WO2024238848A2

WO2024238848A2 - Compositions and methods for managing pesticide resistance

Info

Publication number: WO2024238848A2
Application number: PCT/US2024/029784
Authority: WO
Inventors: Ameer Hamza SHAKEEL; Joseph Frank; Sepehr ZOMORODI; Payam POURTAHERI
Original assignee: Agrospheres Inc
Current assignee: Agrospheres Inc
Priority date: 2023-05-16
Filing date: 2024-05-16
Publication date: 2024-11-21
Anticipated expiration: 2025-11-16
Also published as: WO2024238848A3

Abstract

The present disclosure provides systems, compositions, and methods for restoring one or more pests' susceptibility to pesticides and delaying development of resistance to pesticides. Provided herewith are a composition comprising a minicell comprising (i) a nucleic acid that targets a transcript encoding a polypeptide and/or (ii) a chemical or biological pesticide. Also, provided herewith is a method of managing pesticide resistance that can be developed in pests continuously or repeatedly exposed to pesticides.

Description

COMPOSITIONS AND METHODS FOR MANAGING PESTICIDE RESISTANCE CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of priority to U.S. Provisional Application No. 63/466,889 filed on May 16, 2023, which is hereby incorporated by reference in its entirety. FIELD OF THE DISCLOSURE [0002] The present disclosure generally relates to systems, compositions, and methods for managing pesticide resistance. The present disclosure relates to use of minicell-based RNA for managing pesticide resistance and restoring one or more pests’ susceptibility to pesticides. The present disclosure also relates to systems, compositions, and methods for delaying development of resistance to pesticides using minicell-based RNA. BACKGROUND [0003] Pesticides, largely including insecticides, fungicides, herbicides, molluscicides, avicides, rodenticides, and pediculicides, have been globally utilized to control pests by both chemical and biological methods. Repeated use of the same pesticides can trigger undesirable genetic changes in a pest, which leads to another form of artificial selection, that is pesticide resistance. Pesticide resistance develops due to continuous use of chemical pesticides with the decreased susceptibility of a pest population to a pesticide that was previously effective at controlling the pest. One leading biorational pesticide comes from Bacillus thuringiensis (Bt) identified as insect pathogens, which is Bt insecticidal proteins encoded by the Cry genes. Similarly continuous use of biopesticides or expression of Bt insecticidal proteins in plants can also give rise to selection for resistance in pests. [0004] Management of resistance would be highly desired of companies developing chemical or biological pesticides, as well as farmers and consumers who rely on the commercial pesticides to control pests. Resistance management has been practiced avoiding the development of pesticide resistance such as minimizing pesticide use, avoiding persistent chemicals, avoiding tank mixes, and using long-term rotations of pesticide from different chemical classes. However, the effects of the currently practiced resistance managements are not sufficient to resolve the global issue of pesticide resistance developed in target pest populations. 1 303137700 [0005] Thus, there is an urgent need to provide composition and methods to restore the decreased susceptibility of a pest population to a pesticide and also break down a vicious cycle of pesticide resistance development. SUMMARY OF THE DISCLOSURE [0006] The present disclosure provides an agricultural composition comprising a first minicell encapsulating a nucleic acid that is capable of inducing RNA interference in an agricultural pest. In some embodiments, the agricultural composition further comprises a second minicell encapsulating the pesticide that is capable of killing or controlling the agricultural pest. [0007] In some embodiments, the nucleic acid reduces resistance to or tolerance of a pesticide in the agricultural pest. [0008] In some embodiments, the nucleic acid is capable of recovering the agricultural pest’s sensitivity or susceptibility to the pesticide. In some embodiments, the nucleic acid is capable of altering expression of a gene responsible for pesticide resistance or tolerance. In some embodiments, the expression of the gene responsible for pesticide resistance or tolerance is downregulated or upregulated. [0009] In some embodiments, the gene responsible for pesticide resistance or tolerance is an ion channel gene, a detoxification gene, a target site resistance gene, or a transporter gene. In some embodiments, the ion channel gene is a gene encoding Ryanodine receptor (RyR) or Voltage-gated sodium channel (VGSC). In some embodiments, the detoxification gene is selected from the group consisting of a gene encoding UDP-glycosyltransferase (UGT), Cytochrome P450 monooxygenase, Esterase, Carboxylesterase (CarE), and Glutathione S- transferase (GST). In some embodiments, the target site resistance gene is selected from the group consisting of a gene encoding Acetylcholinesterase (AChE), Voltage-gated sodium channel (VGSC), Gamma-aminobutyric acid (GABA) receptor, Nicotinic acetylcholine receptor (nAChR), and Glutamate-gated chloride channel (GluCl). In some embodiments, the transporter gene is selected from the group consisting of a gene encoding ATP-binding cassette (ABC) transporter, Solute carrier (SLC) transporter, Major facilitator superfamily (MFS) transporter, and P-glycoprotein. [0010] In some embodiments, the agricultural pest is resistant to or tolerant of the pesticide. In some embodiments, the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. 2 303137700 [0011] In some embodiments, the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. In some embodiments, the pesticide is a chemical pesticide or a biological pesticide. In some embodiments, the biological pesticide is a protein toxin. [0012] In some embodiments, the nucleic acid is capable of inducing RNA interference in at least one member from an order selected from the group consisting of: Coleoptera, Diptera, Hymenoptera, Lepidoptera, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Siphonaptera, and Trichoptera. In some embodiments, the nucleic acid is capable of inducing RNA interference in a member of the order Lepidoptera. In some embodiments, the nucleic acid is capable of inducing RNA interference in a member of the genus Plutella. In some embodiments, the nucleic acid is capable of inducing RNA interference in a member of the genus Spodoptera. [0013] In some embodiments, the nucleic acid is a RNA molecule. In some embodiments, the nucleic acid is at least one selected from the group consisting of: a double-stranded RNA (dsRNA) a short-hairpin RNA (shRNA), a small-interfering RNA (siRNA), and a microRNA (miRNA). [0014] In some embodiments, the minicell is ribonuclease deficient. In some embodiments, the minicell comprises at least one fusion protein. In some embodiments, the minicell comprises at least one fusion protein expressed on the surface of the minicell. In some embodiments, the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising at least one target cell adhesion moiety. In some embodiments, the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising a carbohydrate binding molecule. [0015] In some embodiments, the agricultural composition further comprises a solid, dry, or liquid carrier. In some embodiments, said solid carrier is in a form of granule or pellet and is selected from the group consisting of: diatomaceous earth, loam, silica, alginate, clay, bentonite, vermiculite, and combinations thereof. In some embodiments, said dry carrier in a form of powder and is selected from the group consisting of: peat, wheat, bran, vermiculite, clay mineral, calcium carbonate, dolomite, gypsum, bentonite, rock phosphate, phosphorous compound, titanium dioxide, humus, talc, alginate, activated charcoal, and combinations thereof. In some embodiments, said liquid carrier is in a form of liquid or emulsion, and is selected from the group consisting of a surfactant, an emulsifier, a crop oil concentrate, a penetrant, and combinations thereof. 3 303137700 [0016] The present disclosure provides an agricultural composition, comprising: a minicell encapsulating (i) a nucleic acid capable of inducing RNA interference in an agricultural pest and (ii) a pesticide capable of killing or controlling the agricultural pest, wherein the nucleic acid reduces resistance to or tolerance of the pesticide in the agricultural pest. [0017] The present disclosure provides a method of reducing or suppressing pesticide resistance in an agricultural pest, the method comprising: applying an agricultural composition taught herein to an agricultural pest, wherein resistance to a pesticide in the agricultural pest is reduced or suppressed after the application of the agricultural composition. In some embodiments, the resistance to the pesticide is reduced at least 10% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. [0018] The present disclosure provides a method of restoring susceptibility of an agricultural pest to a pesticide, the method comprising: applying an agricultural composition of claim 1 or 36 to an agricultural pest, wherein the agricultural pest is restored to be susceptible to a pesticide after the application of the agricultural composition. In some embodiments, the susceptibility to the pesticide is restored at least 10% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. In some embodiments, the agricultural pest applied with the agricultural composition is more sensitive to the pesticide than an agricultural pest unapplied with the agricultural composition. BRIEF DESCRIPTION OF THE FIGURES [0019] Fig. 1 illustrates two separate minicells encapsulating pesticides and functional RNAs, respectively, and combinational use of the pesticides-encapsulated minicells and the functional RNAs-encapsulated minicells. [0020] Fig.2 illustrates development of a minicell encapsulating both pesticides and RNAs. A parental cell produces functional RNAs (such double-stranded RNAs; dsRNAs) and a minicell containing RNAs is derived from the parental cell. Then, the RNAs-encapsulated minicell is loaded with pesticides of interest, which ends up with an isolated minicell encapsulating both pesticides and RNAs. [0021] Fig. 3 presents a bar graph showing efficacy of minicell-mediated RNA on control of diamondback moth (Plutella xylostella) with resistance to Cry1Ac protoxin. (i.e. no-QAGE strain). 4 303137700 [0022] Fig.4 shows actual efficacy test results of minicell-mediated RNA interference (RNAi), which are quantified and presented in Fig. 3. [0023] Fig. 5 presents a bar graph showing efficacy of minicell mediated RNA on control of protoxin-resistant diamondback moth and protoxin-susceptible diamondback moth, depending on fast and slow release of the minicell mediated RNA. [0024] Fig.6 shows actual efficacy test results of minicell-mediated RNA interference (RNAi), which are quantified and presented in Fig. 5. [0025] Fig. 7A presents relative fold change ryanodine receptor 44F-like gene transcript determined by RT-qPCR analysis on diamondback moth treated with minicell mediated dsRNA. Fig. 7B presents relative fold change of UDP-glucuronosyltransferase receptor gene transcript determined by RT-qPCR analysis on diamondback moth treated with minicell mediated dsRNA. [0026] Figs. 8A-8H show natural infestation field study on cabbages treated with different combination of insecticides: Fig. 8A – untreated control; Fig. 8B – Coragen® insecticide + Dyne-Amic surfactant; Fig. 8C – Xentari® biological insecticide + Dyne-Amic surfactant; Fig. 8D – minicell-mediated dsRNA HP High Conc. + Dyne-Amic surfactant; Fig. 8E – minicell-mediated dsRNA Low Conc. + Dyne-Amic surfactant; Fig. 8F – Coragen® + Dyne- Amic treatment on Week 1 followed by minicell-mediated dsRNA Low Conc. treatment on Week 2; Fig. 8G – minicell-mediated dsRNA Low Conc. + Xentari® + Dyne-Amic; Fig. 8H – Xentari® + Dyne-Amic treatment on Week 1 followed by minicell-mediated dsRNA Low Conc. treatment on Week 2. [0027] Fig.9 shows results of efficacy of minicell-mediated RNA on control of fungi (Botrytis cinerea) with resistance to succinate dehydrogenase inhibitor (SDHI). DESCRIPTION OF THE INVENTION [0028] The present disclosure provides application of a minicell platform formulated to deliver RNAs for delaying development of pesticide resistance, for restoring one or more pests’ susceptibility to pesticides before pesticide resistance has developed, and for reducing the developed pesticide resistance. The present disclosure also provides co-application of a minicell-mediated RNA molecules with pesticides, thereby killing and controlling even pests that have developed pesticide resistance. [0029] The present disclosure is generally directed to an agricultural composition comprising a pesticide (or a plurality of pesticides), a nucleic acid (i.e. functional RNAs for RNA interference), or a combination of both the pesticide and the nucleic acid within a minicell platform. Also, provided is an agricultural composition and/or formulation comprising a 5 303137700 minicell comprising a pesticide or a plurality of pesticides, a minicell comprising a nucleic acid including dsRNA, siRNA, miRNA, and antisense RNA, and a minicell composing both a pesticide and a nucleic acid for RNAi. Further, disclosed are methods of managing pesticide resistance in hosts using an agricultural composition or formulation taught herein. Definitions [0030] While the following terms are believed to be well understood by one of ordinary skill in the art, the following definitions are set forth to facilitate explanation of the presently disclosed subject matter. [0031] The term “a” or “an” refers to one or more of that entity, i.e. can refer to a plural referents. As such, the terms “a” or “an”, “one or more” and “at least one” are used interchangeably herein. In addition, reference to “an element” by the indefinite article “a” or “an” does not exclude the possibility that more than one of the elements is present, unless the context clearly requires that there is one and only one of the elements. [0032] As used herein, “industrially suitable” refers to utilization, and applications, of the minicell, in contexts outside of internally administered human host applications, e.g. outside of administered human therapeutics. [0033] As used herein, “agricultural” composition or compound refers to a substance or compound used in agriculture, such as pesticides, herbicides, fertilizers, growth regulators, animal feeds, animal supplements, or veterinary medicines, but not for human uses. In some embodiments, agricultural activities encompass a wide range of practices, including crop production, livestock farming, forestry, aquaculture, and agroforestry. [0034] The term “biologically active” (synonymous with “bioactive”) indicates that a composition or compound itself has a biological effect, or that it modifies, causes, promotes, enhances, blocks, reduces, limits the production or activity of, or reacts with or binds to an endogenous molecule that has a biological effect. A “biological effect” may be but is not limited to one that impacts a biological process in an plant; one that impacts a biological process in a pest, pathogen or parasite; one that generates or causes to be generated a detectable signal; and the like. Biologically active agents, compositions, complexes or compounds may be used in agricultural applications and compositions. Biologically active agents, compositions, complexes or compounds act to cause or stimulate a desired effect upon a plant, an insect, a worm, bacteria, fungi, or virus. Non-limiting examples of desired effects include, for example, preventing, treating or curing a disease or condition in a host suffering therefrom; limiting the growth of or killing a pest, a pathogen or a parasite that infects a host; augmenting the 6 303137700 phenotype or genotype of a host; stimulating a positive response in a plant to germinate, grow vegetatively, bloom, fertilize, produce fruits and/or seeds, and harvest; and controlling a pest to cause a disease or disorder. [0035] In embodiments, “biologically active compounds,” “biologically active agents,” or “biologicals” encompass a nucleic acid such as RNA biomolecule including antisense nucleic acid, dsRNA, shRNA, siRNA, miRNA, ribozyme, and aptamer. [0036] In the context of agricultural applications of the present disclosure, the term “biologically active” indicates that the composition, complex or compound has an activity that impacts vegetative and reproductive growth of a plant in a positive sense, impacts a plant suffering from a disease or disorder in a positive sense and/or impacts a pest, pathogen or parasite in a negative sense. Thus, a biologically active composition, complex or compound may cause or promote a biological or biochemical activity within a plant that is detrimental to the growth and/or maintenance of a pest, pathogen or parasite; or of cells, tissues or organs of a plant that have abnormal growth or biochemical characteristics and/or a pest, a pathogen or a parasite that causes a disease or disorder within a host such as a plant. [0037] As used herein the term “biocontrol” or “biological control” refers to control of pests by interference with their ecological status, as by introducing a natural enemy or a pathogen into the environment. “Biocontrols” are interchangeably used with ‘biocontrol agents” and “biological control agents”, which are most often referred to as antagonists. Successful biological control reduces the population density of the target species. The term “biocontrol” as a biocontrol agent refers to a compound or composition which originates in a biological matter and is effective in the treatment, prevention, amelioration, inhibition, elimination or delaying the onset of at least one of bacterial, fungal, viral, insect, or any other pest infections or infestations and inhibition of spore germination and hyphae growth. It is appreciated that any biocontrol agent is environmentally safe, that it, it is detrimental to the target species, but does not substantially damage other species in a non-specific manner. Furthermore, it is understood that the term “biocontrol agent” or “biocontrol compound” also encompasses the term “biochemical control agent” or “biochemical control compound”. [0038] As used herein the terms “biostimulant”, “biostimulants” or “biostimulant compound” refers to any microorganism or substance based on natural resources, in the form in which it is supplied to the user, applied to plants, seeds or the root environment soil and any other substrate with the intention to stimulate natural processes of plants to benefit their nutrient use efficiency and/or their tolerance to stress, regardless of its nutrients content, or any combination of such substances and/or microorganisms intended for this use. In some embodiments, 7 303137700 biostimulants refer to biologically active compounds a polypeptide, a metabolite, a semiochemical, a hormone, a pheromone, a micronutrient and a nucleic acid such as RNA biomolecule including antisense nucleic acid, dsRNA, shRNA, siRNA, miRNA, ribozyme, and aptamer. [0039] As used herein, “biological pesticides” or “biopesticides” are substances derived from natural materials such as animals, plants, bacteria, and certain minerals. Unlike chemical pesticides, which are synthetically manufactured, biological pesticides leverage the inherent properties of living organisms or their byproducts to control pests. The “biopesticide” or “biopesticides” also refers to a substance or mixture of substances intended for preventing, destroying or controlling any pest. Specifically, the term relates to substances or mixtures which are effective for treating, preventing, ameliorating, inhibiting, eliminating or delaying the onset of bacterial, fungal, viral, insect- or other pest-related infection or infestation, spore germination and hyphae growth. Also used as substances applied to crops either before or after harvest to protect the commodity from deterioration during storage and transport. As a contraction of 'biological pesticides', biopesticides include several types of pest management intervention through predatory, parasitic, or chemical relationships. The term has been associated historically with biological control – and by implication – the manipulation of living organisms. In some embodiments, biopesticides refer to biologically active compounds a polypeptide, a metabolite, a semiochemical, a hormone, a pheromone, a macronutrient, a micronutrient and a nucleic acid such as RNA biomolecule including antisense nucleic acid, dsRNA, shRNA, siRNA, miRNA, ribozyme, and aptamer. In some embodiments, the biological pesticide is a protein toxin, that is a protoxin. [0040] As used herein, “chemical pesticides” are substances that are used to control, repel, or eliminate pests such as insects, weeds, fungi, and rodents that can harm crops or livestock, but not humans. These pesticides are formulated using various chemicals, including synthetic compounds, to target specific pests or pest categories. Chemical pesticides work by interfering with the pest's physiology, behavior, or reproductive system, ultimately reducing their population or preventing damage to crops or property. [0041] The term “pest” is defined herein as encompassing vectors of plant, humans or livestock disease, unwanted species of bacteria, fungi, viruses, insects, nematodes mites, ticks or any organism causing harm during or otherwise interfering with the production, processing, storage, transport or marketing of food, agricultural commodities, wood and wood products or animal feedstuffs. Insect pests include insects selected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera Orthroptera, Thysanoptera, 8 303137700 Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, etc., particularly Lepidoptera and Coleoptera. Those skilled in the art will recognize that not all compounds are equally effective against all pests. Compounds of the embodiments display activity against insect pests, which may include economically important agronomic, forest, greenhouse, nursery ornamentals, food and fiber, public and animal health, domestic and commercial structure, household and stored product pests. [0042] As used herein the terms “cellular organism” “microorganism” or “microbe” should be taken broadly. These terms are used interchangeably and include, but are not limited to, the two prokaryotic domains, Bacteria and Archaea, as well as certain eukaryotic fungi and protists. [0043] The term “prokaryotes” is art recognized and refers to cells that contain no nucleus or other cell organelles. The prokaryotes are generally classified in one of two domains, the Bacteria and the Archaea. The definitive difference between organisms of the Archaea and Bacteria domains is based on fundamental differences in the nucleotide base sequence in the 16S ribosomal RNA. [0044] The term “Archaea” refers to a categorization of organisms of the division Mendosicutes, typically found in unusual environments and distinguished from the rest of the prokaryotes by several criteria, including the number of ribosomal proteins and the lack of muramic acid in cell walls. On the basis of ssrRNA analysis, the Archaea consist of two phylogenetically-distinct groups: Crenarchaeota and Euryarchaeota. On the basis of their physiology, the Archaea can be organized into three types: methanogens (prokaryotes that produce methane); extreme halophiles (prokaryotes that live at very high concentrations of salt (NaCl); and extreme (hyper) thermophilus (prokaryotes that live at very high temperatures). Besides the unifying archaeal features that distinguish them from Bacteria (i.e., no murein in cell wall, ester-linked membrane lipids, etc.), these prokaryotes exhibit unique structural or biochemical attributes which adapt them to their particular habitats. The Crenarchaeota consists mainly of hyperthermophilic sulfur-dependent prokaryotes and the Euryarchaeota contains the methanogens and extreme halophiles. [0045] “Bacteria” or “eubacteria” refers to a domain of prokaryotic organisms. Bacteria include at least 11 distinct groups as follows: (1) Gram-positive (gram+) bacteria, of which there are two major subdivisions: (1) high G+C group (Actinomycetes, Mycobacteria, Micrococcus, others) (2) low G+C group (Bacillus, Clostridia, Lactobacillus, Staphylococci, Streptococci, Mycoplasmas); (2) Proteobacteria, e.g., Purple photosynthetic+non- photosynthetic Gram-negative bacteria (includes most “common” Gram-negative bacteria); (3) 9 303137700 Cyanobacteria, e.g., oxygenic phototrophs; (4) Spirochetes and related species; (5) Planctomyces; (6) Bacteroides, Flavobacteria; (7) Chlamydia; (8) Green sulfur bacteria; (9) Green non-sulfur bacteria (also anaerobic phototrophs); (10) Radioresistant micrococci and relatives; (11) Thermotoga and Thermosipho thermophiles. [0046] A “eukaryote” is any organism whose cells contain a nucleus and other organelles enclosed within membranes. Eukaryotes belong to the taxon Eukarya or Eukaryota. The defining feature that sets eukaryotic cells apart from prokaryotic cells (the aforementioned Bacteria and Archaea) is that they have membrane-bound organelles, especially the nucleus, which contains the genetic material, and is enclosed by the nuclear envelope. [0047] The terms “genetically modified host cell,” “recombinant host cell,” and “recombinant strain” are used interchangeably herein and refer to host cells that have been genetically modified by the cloning and transformation methods of the present disclosure. Thus, the terms include a host cell (e.g., bacteria, yeast cell, fungal cell, CHO, human cell, etc.) that has been genetically altered, modified, or engineered, such that it exhibits an altered, modified, or different genotype and/or phenotype (e.g., when the genetic modification affects coding nucleic acid sequences of the microorganism), as compared to the naturally-occurring organism from which it was derived. It is understood that in some embodiments, the terms refer not only to the particular recombinant host cell in question, but also to the progeny or potential progeny of such a host cell. [0048] The term “wild-type microorganism” or “wild-type host cell” describes a cell that occurs in nature, i.e. a cell that has not been genetically modified. In the disclosure, “wild type strain” or “wild strain” or “wild type cell line” refers to a cell strain/line that can produce minicells. In some embodiments, wild type bacterial strains and/or cell lines such as E. coli strain p678-54 and B. subtilis strain CU403 can make miniature cells deficient in DNA. Methods for producing such minicells are known in the art. See, for example, Adler et al., 1967, Proc. Natl. Acad. Sci. USA 57:321-326; Inselburg J, 1970 J. Bacteriol.102(3):642-647; Frazer 1975, Curr. Topics Microbiol. Immunol. 69:1-84, Reeve et al 1973, J. Bacteriol. 114(2):860- 873; and Mendelson et al 1974 J. Bacteriol. 117(3):1312-1319. [0049] The term “genetically engineered” may refer to any manipulation of a host cell’s genome (e.g. by insertion, deletion, mutation, or replacement of nucleic acids). [0050] The term “control host cell” refers to an appropriate comparator host cell for determining the effect of a genetic modification or experimental treatment. In some embodiments, the control host cell is a wild type cell. In other embodiments, a control host cell 10 303137700 is genetically identical to the genetically modified host cell, save for the genetic modification(s) differentiating the treatment host cell. [0051] As used herein, the term “genetically linked” refers to two or more traits that are co- inherited at a high rate during breeding such that they are difficult to separate through crossing. [0052] A “recombination” or “recombination event” as used herein refers to a chromosomal crossing over or independent assortment. [0053] As used herein, the term “phenotype” refers to the observable characteristics of an individual cell, cell culture, organism, or group of organisms which results from the interaction between that individual’s genetic makeup (i.e., genotype) and the environment. [0054] As used herein, the term “chimeric” or “recombinant” when describing a nucleic acid sequence or a protein sequence refers to a nucleic acid, or a protein sequence, that links at least two heterologous polynucleotides, or two heterologous polypeptides, into a single macromolecule, or that rearranges one or more elements of at least one natural nucleic acid or protein sequence. For example, the term “recombinant” can refer to an artificial combination of two otherwise separated segments of sequence, e.g., by chemical synthesis or by the manipulation of isolated segments of nucleic acids by genetic engineering techniques. [0055] As used herein, a “synthetic nucleotide sequence” or “synthetic polynucleotide sequence” is a nucleotide sequence that is not known to occur in nature or that is not naturally occurring. Generally, such a synthetic nucleotide sequence will comprise at least one nucleotide difference when compared to any other naturally occurring nucleotide sequence. [0056] As used herein, a “synthetic amino acid sequence” or “synthetic peptide” or “synthetic protein” is an amino acid sequence that is not known to occur in nature or that is not naturally occurring. Generally, such a synthetic protein sequence will comprise at least one amino acid difference when compared to any other naturally occurring protein sequence. [0057] As used herein, the term “nucleic acid” refers to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides, or analogs thereof. This term refers to the primary structure of the molecule, and thus includes double- and single-stranded DNA, as well as double- and single-stranded RNA. It also includes modified nucleic acids such as methylated and/or capped nucleic acids, nucleic acids containing modified bases, backbone modifications, and the like. The terms “nucleic acid” and “nucleotide sequence” are used interchangeably. [0058] As used herein, the term “gene” refers to any segment of DNA associated with a biological function. Thus, genes include, but are not limited to, coding sequences and/or the regulatory sequences required for their expression. Genes can also include non-expressed DNA 11 303137700 segments that, for example, form recognition sequences for other proteins. Genes can be obtained from a variety of sources, including cloning from a source of interest or synthesizing from known or predicted sequence information, and may include sequences designed to have desired parameters. [0059] As used herein, the term “homologous” or “homologue” or “ortholog” is known in the art and refers to related sequences that share a common ancestor or family member and are determined based on the degree of sequence identity. The terms “homology,” “homologous,” “substantially similar” and “corresponding substantially” are used interchangeably herein. They refer to nucleic acid fragments wherein changes in one or more nucleotide bases do not affect the ability of the nucleic acid fragment to mediate gene expression or produce a certain phenotype. These terms also refer to modifications of the nucleic acid fragments of the instant disclosure such as deletion or insertion of one or more nucleotides that do not substantially alter the functional properties of the resulting nucleic acid fragment relative to the initial, unmodified fragment. It is therefore understood, as those skilled in the art will appreciate, that the disclosure encompasses more than the specific exemplary sequences. These terms describe the relationship between a gene found in one species, subspecies, variety, cultivar or strain and the corresponding or equivalent gene in another species, subspecies, variety, cultivar or strain. For purposes of this disclosure homologous sequences are compared. “Homologous sequences” or “homologues” or “orthologs” are thought, believed, or known to be functionally related. A functional relationship may be indicated in any one of a number of ways, including, but not limited to: (a) degree of sequence identity and/or (b) the same or similar biological function. Preferably, both (a) and (b) are indicated. Homology can be determined using software programs readily available in the art, such as those discussed in Current Protocols in Molecular Biology (F.M. Ausubel et al., eds., 1987) Supplement 30, section 7.718, Table 7.71. [0060] As used herein, the term “endogenous” or “endogenous gene,” refers to the naturally occurring gene, in the location in which it is naturally found within the host cell genome. In the context of the present disclosure, operably linking a heterologous promoter to an endogenous gene means genetically inserting a heterologous promoter sequence in front of an existing gene, in the location where that gene is naturally present. An endogenous gene as described herein can include alleles of naturally occurring genes that have been mutated according to any of the methods of the present disclosure. [0061] As used herein, the term “exogenous” is used interchangeably with the term “heterologous,” and refers to a substance coming from some source other than its native source. For example, the terms “exogenous protein,” or “exogenous gene” refer to a protein or gene 12 303137700 from a non-native source or location, and that have been artificially supplied to a biological system. [0062] As used herein, the term “nucleotide change” refers to, e.g., nucleotide substitution, deletion, and/or insertion, as is well understood in the art. For example, mutations contain alterations that produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded protein or how the proteins are made. [0063] As used herein, the term “protein modification” refers to, e.g., amino acid substitution, amino acid modification, deletion, and/or insertion, as is well understood in the art. [0064] As used herein, the term “at least a portion” or “fragment” of a nucleic acid or polypeptide means a portion having the minimal size characteristics of such sequences, or any larger fragment of the full length molecule, up to and including the full length molecule. A fragment of a polynucleotide of the disclosure may encode an enzymatically active portion of a genetic regulatory element. An enzymatically active portion of a genetic regulatory element can be prepared by isolating a portion of one of the polynucleotides of the disclosure that comprises the genetic regulatory element and assessing activity as described herein. Similarly, a portion of a polypeptide may be 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, and so on, going up to the full length polypeptide. The length of the portion to be used will depend on the particular application. A portion of a nucleic acid useful as a hybridization probe may be as short as 12 nucleotides; in some embodiments, it is 20 nucleotides. A portion of a polypeptide useful as an epitope may be as short as 4 amino acids. A portion of a polypeptide that performs the function of the full-length polypeptide would generally be longer than 4 amino acids. [0065] Variant polynucleotides also encompass sequences derived from a mutagenic and recombinogenic procedure such as DNA shuffling. Strategies for such DNA shuffling are known in the art. See, for example, Stemmer (1994) PNAS 91:10747-10751; Stemmer (1994) Nature 370:389-391; Crameri et al.(1997) Nature Biotech. 15:436-438; Moore et al.(1997) J. Mol. Biol.272:336-347; Zhang et al.(1997) PNAS 94:4504-4509; Crameri et al.(1998) Nature 391:288-291; and U.S. Patent Nos. 5,605,793 and 5,837,458. [0066] For PCR amplifications of the polynucleotides disclosed herein, oligonucleotide primers can be designed for use in PCR reactions to amplify corresponding DNA sequences from cDNA or genomic DNA extracted from any organism of interest. Methods for designing PCR primers and PCR cloning are generally known in the art and are disclosed in Sambrook et al.(2001) Molecular Cloning: A Laboratory Manual (3^rd ed., Cold Spring Harbor Laboratory Press, Plainview, New York). See also Innis et al., eds. (1990) PCR Protocols: A Guide to 13 303137700 Methods and Applications (Academic Press, New York); Innis and Gelfand, eds. (1995) PCR Strategies (Academic Press, New York); and Innis and Gelfand, eds. (1999) PCR Methods Manual (Academic Press, New York). Known methods of PCR include, but are not limited to, methods using paired primers, nested primers, single specific primers, degenerate primers, gene-specific primers, vector-specific primers, partially-mismatched primers, and the like. [0067] The term “primer” as used herein refers to an oligonucleotide which is capable of annealing to the amplification target allowing a DNA polymerase to attach, thereby serving as a point of initiation of DNA synthesis when placed under conditions in which synthesis of primer extension product is induced, i.e., in the presence of nucleotides and an agent for polymerization such as DNA polymerase and at a suitable temperature and pH. The (amplification) primer is preferably single stranded for maximum efficiency in amplification. Preferably, the primer is an oligodeoxyribonucleotide. The primer must be sufficiently long to prime the synthesis of extension products in the presence of the agent for polymerization. The exact lengths of the primers will depend on many factors, including temperature and composition (A/T vs. G/C content) of primer. A pair of bi-directional primers consists of one forward and one reverse primer as commonly used in the art of DNA amplification such as in PCR amplification. [0068] As used herein, “promoter” refers to a DNA sequence capable of controlling the expression of a coding sequence or functional RNA. In some embodiments, the promoter sequence consists of proximal and more distal upstream elements, the latter elements often referred to as enhancers. Accordingly, an “enhancer” is a DNA sequence that can stimulate promoter activity, and may be an innate element of the promoter or a heterologous element inserted to enhance the level or tissue specificity of a promoter. Promoters may be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even comprise synthetic DNA segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental conditions. It is further recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, DNA fragments of some variation may have identical promoter activity. [0069] As used herein, the phrases “recombinant construct”, “expression construct”, “chimeric construct”, “construct”, and “recombinant DNA construct” are used interchangeably herein. Also, “construct”, “vector”, and “plasmid” are used interchangeably herein. A recombinant construct comprises an artificial combination of nucleic acid fragments, e.g., 14 303137700 regulatory and coding sequences that are not found together in nature. For example, a chimeric construct may comprise regulatory sequences and coding sequences that are derived from different sources, or regulatory sequences and coding sequences derived from the same source, but arranged in a manner different than that found in nature. Such construct may be used by itself or may be used in conjunction with a vector. If a vector is used then the choice of vector is dependent upon the method that will be used to transform host cells. For example, a plasmid vector can be used. The skilled artisan is well aware of the genetic elements that must be present on the vector in order to successfully transform, select and propagate host cells comprising any of the isolated nucleic acid fragments of the disclosure. The skilled artisan will also recognize that different independent transformation events will result in different levels and patterns of expression (Jones et al., (1985) EMBO J. 4:2411-2418; De Almeida et al., (1989) Mol. Gen. Genetics 218:78-86), and thus that multiple events must be screened in order to obtain lines displaying the desired expression level and pattern. Such screening may be accomplished by Southern analysis of DNA, Northern analysis of mRNA expression, immunoblotting analysis of protein expression, or phenotypic analysis, among others. Vectors can be plasmids, viruses, bacteriophages, pro-viruses, phagemids, transposons, artificial chromosomes, and the like, that replicate autonomously or can integrate into a chromosome of a host cell. A vector can also be a naked RNA polynucleotide, a naked DNA polynucleotide, a polynucleotide composed of both DNA and RNA within the same strand, a poly-lysine- conjugated DNA or RNA, a peptide-conjugated DNA or RNA, a liposome-conjugated DNA, or the like, that is not autonomously replicating. As used herein, the term “expression” refers to the production of a functional end-product e.g., an mRNA or a protein (precursor or mature). [0070] “Operably linked” means in this context the sequential arrangement of the promoter polynucleotide according to the disclosure with a further oligo- or polynucleotide, resulting in transcription of said further polynucleotide. [0071] As used herein, the term "display" refers to the exposure of the polypeptide of interest on the outer surface of the minicell. By way of non-limiting example, the displayed polypeptide may be a protein or a protein domain which is either expressed on the minicell membrane or is associated with the minicell membrane such that the extracellular domain or domain of interest is exposed on the outer surface of the minicell (expressed and displayed on the surface of the minicell or expressed in the parental cell to be displayed on the surface of the segregated/budded minicell). In all instances, the "displayed" protein or protein domain is available for interaction with extracellular components. A membrane-associated protein may 15 303137700 have more than one extracellular domain, and a minicell of the disclosure may display more than one membrane-associated protein. [0072] As used herein, the terms "polypeptide", "protein" and "protein domain" refer to a macromolecule made up of a single chain of amino acids joined by peptide bonds. Polypeptides of the invention may comprise naturally occurring amino acids, synthetic amino acids, genetically encoded amino acids, non-genetically encoded amino acids, and combinations thereof. Polypeptides may include both L-form and D-form amino acids. [0073] As used herein, the term "enzymatically active polypeptide" refers to a polypeptide which encodes an enzymatically functional protein. The term "enzymatically active polypeptide" includes but not limited to fusion proteins which perform a biological function. Exemplary enzymatically active polypeptides, include but not limited to enzymes/enzyme moiety (e.g. wild type, variants, or engineered variants) that specifically bind to certain receptors or biological/chemical substrates to effect a biological function such as biological signal transduction or chemical inactivation. [0074] As used herein, the term "protease-deficient strain" refers to a strain that is deficient in one or more endogenous proteases. For example, protease deficiency can be created by deleting, removing, knock-out, silencing, suppressing, or otherwise downregulating at lease on endogenous protease. Said proteases can include catastrophic proteases. For example, BL21 (DE3) E. coli strain is deficient in proteases Lon and OmpT. E. coli strain has cytoplasmic proteases and membrane proteases that can significantly decrease protein production and localization to the membrane. In some embodiments, a protease-deficient strain can maximize production and localization of a protein of interest to the membrane of the cell. “Protease- deficient” can be interchangeably used as “protease-free” in the present disclosure. [0075] As used herein, the term "ribonuclease-deficient strain" refers to a strain that is deficient in one or more endogenous ribonuclease. For example, ribonuclease deficiency can be created by deleting, removing, knock-out, silencing, suppressing, or otherwise downregulating at lease on endogenous ribonuclease. Said ribonuclease can include ribonuclease III. For example, HT115 E. coli strain is deficient in RNase III. In some embodiments, a ribonuclease-deficient strain is unable to and/or has a reduced capability of recognizing dsRNA and cleaving it at specific targeted locations. “Ribonuclease-deficient” can be interchangeably used as “ribonuclease-free” in the present disclosure. [0076] As used herein, the term "anucleated cell" refers to a cell that lacks a nucleus and also lacks chromosomal DNA and which can also be termed as an “anucleate cell”. Because eubacterial and archaebacterial cells, unlike eukaryotic cells, naturally do not have a nucleus 16 303137700 (a distinct organelle that contains chromosomes), these non-eukaryotic cells are of course more accurately described as being "without chromosomes" or "achromosomal." Nonetheless, those skilled in the art often use the term "anucleated" when referring to bacterial minicells in addition to other eukaryotic minicells. Accordingly, in the present disclosure, the term "minicells" encompasses derivatives of eubacterial cells that lack a chromosome; derivatives of archaebacterial cells that lack their chromosome(s), and anucleate derivatives of eukaryotic cells that lack a nucleus and consequently a chromosome. Thus, in the present disclosure, "anucleated cell" or "anucleate cell" can be interchangeably used with the term “achromosomal cell” and “minicell.” [0077] “Sequence identity" or "identity" in the context of two nucleic acid or polypeptide sequences includes reference to the number of residues in the two sequences which are the same when aligned for maximum correspondence over a specified comparison window. When percentage of sequence identity is used in reference to proteins it is recognized that residue positions which are not identical often differ by conservative amino acid substitutions, where amino acid residues are substituted for other amino acid residues with similar chemical properties (e.g., charge or hydrophobicity) and therefore do not change the functional properties of the molecule. Where sequences differ in conservative substitutions, the percent sequence identity may be adjusted upwards to correct for the conservative nature of the substitution. Sequences which differ by such conservative substitutions are said to have "sequence similarity" or "similarity." Means for making this adjustment are well-known to those of skill in the art. Typically this involves scoring a conservative substitution as a partial rather than a full mismatch, thereby increasing the percentage sequence identity. Thus, for example, where an identical amino acid is given a score of 1 and a non-conservative substitution is given a score of zero, a conservative substitution is given a score between zero and 1. The scoring of conservative substitutions is calculated, e.g., according to the algorithm of Meyers and Miller, Computer Applic. Biol. Sci., 4:11-17 (1988). The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. An example of a local alignment algorithm utilized for the comparison of sequences is the NCBI Basic Local Alignment Search Tool (BLAST®) (Altschul et al. 1990 J. Mol. Biol. 215: 403-10), which is available from several sources, including the National Center for Biotechnology Information (NCBI, Bethesda, Md.) and on the Internet, for use in connection with the sequence analysis programs blastp, blastn, blastx, tblastn and tblastx. It can be accessed on the internet via the National Library of Medicine (NLM)'s world-wide-web URL. A description of how to determine sequence identity using this 17 303137700 program is available at the NLM's website on BLAST tutorial. Another example of a mathematical algorithm utilized for the global comparison of sequences is the Clustal W and Clustal X (Larkin et al. 2007 Bioinformatics, 23, 2947-294, Clustal W and Clustal X version 2.0) as well as Clustal omega. Unless otherwise stated, references to sequence identity used herein refer to the NCBI Basic Local Alignment Search Tool (BLAST®). [0078] As used herein, the term “binding site,” means a molecular structure or compound, such as a protein, a polypeptide, a polysaccharide, a glycoprotein, a lipoprotein, a fatty acid, a lipid or a nucleic acid or a particular region in such molecular structure or compound or a particular conformation of such molecular structure or compound, or a combination or complex of such molecular structures or compounds. In certain embodiments, at least one binding site is on an intact living plant. An “intact living plant,” as used herein, means a plant as it grows, whether it grows in soil, in water or in artificial substrate, and whether it grows in the field, in a greenhouse, in a yard, in a garden, in a pot or in hydroponic culture systems. An intact living plant preferably comprises all plant parts (roots, stem, branches, leaves, needles, thorns, flowers, seeds etc.) that are normally present on such plant in nature, although some plant parts, such as, e.g., flowers, may be absent during certain periods in the plant's life cycle. [0079] A “binding domain,” as used herein, means the whole or part of a proteinaceous (protein, protein-like or protein containing) molecule that is capable of binding using specific intermolecular interactions to a target molecule. A binding domain can be a naturally occurring molecule, it can be derived from a naturally occurring molecule, or it can be entirely artificially designed. A binding domain can be based on domains present in proteins, including but not limited to microbial proteins, antibodies, enzymes, protease inhibitors, protein toxins, fibronectin, lipocalins, single-chain antiparallel coiled coil proteins or repeat motif proteins. Non-limiting examples of such binding domains are carbohydrate binding modules (CBM) such as cellulose binding domain to be targeted to plants, ACC-deaminase, cutinase, cellulose and the like. In some embodiments, a cell adhesion moiety comprises a binding domain. In other embodiments, a cell stimulation moiety comprises a binding domain. In further embodiments, a cell degradation moiety comprises a binding domain. [0080] As used herein, “carrier,” “acceptable carrier,” or “biologically actively acceptable carrier” refers to a diluent, adjuvant, excipient, surfactant, or vehicle with which a composition can be administered to its target, which does not detrimentally effect the composition. [0081] In some embodiments, biologically active compounds can be used as biocontrols and biostimulants that have become the new age of crop protection and enhancement. 18 303137700 [0082] An example of a biocontrol is RNA molecules capable of inducing RNA interference (RNAi), which is used to repress, silence, knock down or downregulate target gene expression, and/or delay gene expression or translation of mRNA. Also, RNAi is capable of activating or upregulating target gene expression for transcript activation. The RNAi can kill target pest by specifically repressing target gene transcripts, while leaving the non targeted pests unharmed. In invertebrates, long dsRNA can be efficiently used to silence gene expression without activation of dsRNA-activated protein kinase (PKR) or the interferon response that has been shown to occur in mammalian cell systems. [0083] The present disclosure teaches minicell-mediated RNAi or minicell-based RNAi, in which the minicells of the present disclosure encapsulate and deliver a nucleic acid (e.g., RNA molecule such as a double-stranded RNA (dsRNA), a short-hairpin RNA (shRNA), a small- interfering RNA (siRNA), and a microRNA (miRNA)) that is capable of inducing RNAi in an agricultural target pest. [0084] Another example of biologically active compounds as a biocontrol is protein toxins or protoxins. Protein toxins, also known as protoxins, are biologically active substances produced by certain organisms, including bacteria, plants, and animals. These toxins are proteins or polypeptides that can cause harm to other organisms by disrupting cellular functions or structures, thereby leading to physiological disturbances, disease, or death in the affected organisms. Examples of protein toxins/protoxins include botulinum toxin produced by Clostridium botulinum bacteria, ricin produced by the castor bean plant, and cry toxins produced by Bacillus thuringiensis (Bt). [0085] The cry toxins are highly beneficial for managing insect pests across various crops, forests, and disease-carrying insects affecting humans. However, the emergence of resistance in insect populations poses a significant challenge to the continued efficacy of this environmentally friendly insecticide. The primary mode of resistance in insects against Cry toxins is characterized by a diminished ability of the toxins to bind to the specific receptors found in the insects' digestive tract. (Ferre and Van Rie, 2002). [0086] Plant antibodies are another form of biocontrols that can be used to specifically target pests. Immunoglobulin domains, light chain, heavy chain, and CDRs, Fv, Fab, and Fc regions can be encapsulated as active compounds and be delivered to a target. The present disclosure provides fungicidal antibodies such as those generated from glucosylceramide. [0087] Plant-growth regulators, hormones, enzymes, pheromones, allomones and kairomones are also biocontrols. A pheromone can act as a biocontrol to prevent bugs and/or insects from mating. 19 303137700 [0088] Biostimulants foster plant development in a number of demonstrated ways throughout the crop lifecycle, from seed germination to plant maturity. They can be applied to plant, seed, soil or other growing media that may enhance the plant’s ability to assimilate nutrients and properly develop. By fostering complementary soil microbes and improving metabolic efficiency, root development and nutrient delivery, biostimulants can increase yield in terms of weight, seed and fruit set, enhance quality, affecting sugar content, color and shelf life, improve the efficiency of water usage, and strengthen stress tolerance and recovery. These biostimulants can include pheromones or enzymes like ACC-Deaminase. [0089] Biostimulants are compounds that produce non-nutritional plant growth responses and reduce stress by enhancing stress tolerance. Fertilizers, which produce a nutritional response can be considered as biostimulants. Many important benefits of biostimulants are based on their ability to influence hormonal activity. Hormones in plants (phytohormones) are chemical messengers regulating normal plant development as well as responses to the environment. Root and shoot growth, as well as other growth responses are regulated by phytohormones. Compounds in biostimulants can alter the hormonal status of a plant and exert large influences over its growth and health. Sea kelp, humic acids and B Vitamins are common components of biostimulants that are important sources of compounds that influence plant growth and hormonal activity. Antioxidants are another group of plant chemicals that are important in regulating the plants response to environmental and chemical stress (drought, heat, UV light and herbicides). When plants come under stress, “free radicals” or reactive oxygen molecules (e.g., hydrogen peroxide) damage the plants cells. Antioxidants suppress free radical toxicity. Plants with the high levels of antioxidants produce better root and shoot growth, maintain higher leaf-moisture content and lower disease incidence in both normal and stressful environments. Applying a biostimulant enhances antioxidant activity, which increases the plant's defensive system. Vitamin C, Vitamin E, and amino acids such as glycine are antioxidants contained in biostimulants. [0090] Biostimulants may act to stimulate the growth of microorganisms that are present in soil or other plant growing medium. Biostimulants are capable of stimulating growth of microbes included in the microbial inoculant. Thus, it is desirable to obtain a biostimulant, that, when used with a microbial inoculant, is capable of enhancing the population of both native microbes and inoculant microbes. [0091] As used herein, “agrochemicals” also known as “agricultural chemicals,” are substances used in agriculture to enhance crop production, protect crops from pests and diseases, and manage weeds. These chemicals play a crucial role in modern agriculture by improving crop 20 303137700 yields, quality, and overall profitability. Main types of agrochemicals and their functions are as follows; (1) Pesticides are chemicals used to control pests, including insects, weeds, fungi, and pathogens, that can damage crops and reduce yields. They include insecticides (for insects), herbicides (for weeds), fungicides (for fungi), and bactericides (for bacteria). Pesticides can be applied through spraying, dusting, or seed treatment; (2) fertilizers provide essential nutrients to crops to promote healthy growth and maximize yields. They typically contain nitrogen, phosphorus, potassium, and other micronutrients necessary for plant growth. Fertilizers can be applied to the soil or sprayed directly onto the foliage; (3) Plant Growth Regulators (PGRs) are chemicals that influence plant growth and development. They can be used to stimulate or inhibit various physiological processes in plants, such as flowering, fruit ripening, and stem elongation. PGRs are commonly used in horticulture and to improve the quality of fruit and ornamental crops; (4) Soil amendments are substances added to soil to improve its physical, chemical, or biological properties. They include lime, gypsum, compost, and organic matter, which can help enhance soil fertility, structure, and water retention capacity; (5) Adjuvants are additives used in conjunction with pesticides to improve their effectiveness and performance. They can enhance pesticide uptake, adhesion, and spread on plant surfaces, as well as reduce spray drift and increase rainfastness. [0092] In some embodiments, the present disclosure provides a minicell encapsulating and delivering a nucleic acid capable of inducing RNAi and/or a chemical or biological pesticide in a scalable, cost-effective manner. The minicell and/or a composition comprising the minicell and a plurality of substances (e.g., a nucleic acid capable of inducing RNAi and/or a chemical or biological pesticide) can also be modified to invasively deliver the plurality of substances to a target or locus of interest. [0093] In one aspect, the minicell and/or a composition comprising the minicell are derived from bacterial cells lacking ribonucleases (ribonuclease III) and has T7, T3 or Sp6 RNA polymerase promoters to produce dsRNA used for RNA interference (RNAi) of a target. This bacterial cell is then modified to produce minicells with the dsRNA encapsulated within them. This helps simplify and cheapen purification and encapsulation. By encapsulating dsRNA, the dsRNA molecules are protected from environmental RNases. For examples, pests including insects orally consume the minicells for the delivery of the dsRNA. Once inside the insects, dsRNAs are a substrate for RNase III-like proteins referred to as Dicer or Dicer-like proteins. Dicer appears to preferentially initiate dsRNA cleavage at the ends of the dsRNA, making successive cleavages to generate 21- to 24-bp small-interfering (si) RNA duplexes to silence and/or suppress their target transcripts and inhibit translations of the transcripts. The resulting 21 303137700 siRNA duplexes are loaded into a multiprotein complex called the RNA-induced silencing complex (RISC) where the passenger (sense) strand is removed and the guide (antisense) strand remains to target mRNA for silencing. The guide strand in the RISC enables base pairing of the complex to complementary mRNA transcripts and enzymatic cleavage of the target mRNA by a class of proteins referred to as Argonaute proteins, thereby preventing translation of the target mRNA. This is what causes the death of the targeted pest, while leaving untargeted pests unharmed. Also, the minicell and/or a composition comprising the minicell can be utilized to encapsulate dsRNA, siRNA shRNA, or miRNA. In other aspects, antisense nucleic acid, ribozyme, or aptamer can be encapsulated within the minicell. [0094] In some embodiments, the minicell and the dsRNA are produced from different host cells and are incubated together after the independent productions have been completed. In some embodiments, the minicells can be utilized to internally express dsRNA from a recombinant plasmid capable of producing dsRNA inside of the anucleate minicell. Then, the internally produced dsRNA is delivered to its target within the anucleate minicell. In other embodiments, the minicells can be utilized to encapsulate externally and/or exogenously produced dsRNA that is first produced outside of the anucleate minicell. Then, the externally- produced dsRNA encapsulated into the minicell is delivered to its target within the anucleate minicell. In further embodiments, the minicells can be utilized to internally express dsRNA within the platform and encapsulate one or more sequences of exogenously-produced dsRNA into the platform for the purposes of targeting one or multiple different pests. This entails encapsulating dsRNA that is either homologous or heterologous to the internally expressed dsRNA sequence in the anucleate cell. Thus, the minicell can carry both internally-expressed dsRNA and externally-expressed, but encapsulated dsRNA over to its intended target. [0095] The present disclosure teaches that the minicells and/or an agricultural composition can deliver internally-produced dsRNA and externally/exogenously-produced dsRNA individually, or together to a target cell. The target cell is not a mammalian cell. [0096] The present disclosure teaches that an industrially suitable minicell and/or an agricultural composition for encapsulation and delivery of at least one biologically active compound, comprising: an intact anucleated cell derived from a ribonuclease deficient parental cell, comprising at least one biologically active compound within said cell, wherein said biologically active compound is a nucleic acid, wherein the nucleic acid targets a transcript encoding a polypeptide within a target cell, and wherein the target cell is not a mammalian cell. The minicell and/or an agricultural composition further comprises at least one biologically acceptable carrier. 22 303137700 [0097] In some embodiments, for protein-mediated biocontrols, the present disclosure uses bacterial cells lacking proteases and has T7, T3, or Sp6 polymerase promoters to produce a significant amount of proteins. This bacterial cell is then modified to produce minicells with the proteins immobilized to their surface or encapsulated within them. A protein-expressing plasmid is integrated into the nucleoid DNA of the bacteria to safely and efficiently produce proteins. Insects then interact with or orally consume the minicells that express or retain the desired proteins. For antibody-mediated biocontrols, minicells can express or encapsulate antibodies to specifically target unwanted pests. Minicells can deliver antibodies or recombinant antibodies that serve as highly specific biopesticides against insects or fungal pathogens (Raymond et al., Fungal Biology Review 25(2) :84-88, 2011). [0098] In some embodiments, for biostimulants, the present disclosure teaches that minicells can deliver a wide range of plant -growth promoting biomolecules to the surface of the plant, its seeds, and its root system. Many of these biomolecules occur as a result of a dynamic, symbiotic relationship that some microorganisms have with plants and are produced naturally in response to certain environmental cues or stresses. The minicell can be engineered to deliver a high-payload capacity of these plant growth promoting biomolecules, either immobilized extracellularly on their surface or encapsulated intracellularly, without relying on microorganism or plants to naturally produce them. This enables a higher effective concentration of these biomolecules to be delivered to the plant microenvironment while also allowing for a more controlled, adaptive response to agricultural input needs. Many of these biomolecules are enzymes that bacteria produce, either intracellularly or extracellularly, that play an important role in promoting soil fertility and providing defense against plant pathogens (Jog et al, Journal of Applied Microbiology 113:1154-1164, 2012; Sathya et al. 3 Biotech 7:102, 2017). Others, like 1-aminocyclopropane-1-carboxylate (ACC) Deaminase, can regulate plant growth on a hormonal level by lowering ethylene levels in the plant microenvironment (Souza et al., Genet. Mol. Biol. 38(4): 401-419, 2015) . [0099] In some embodiments, the biologically active compound are valuable enzymes that could be produced and delivered to the plant or its root system using the minicell, which include, but are not limited to cellulase, phytase, chitinase, protease, phosphatase, nucleases, lipases, glucanases, xylanases, amylases, peptidases, peroxidases, ligninases, pectinases, hemicellulases, and keratinases. Beyond being able to effectively deliver enzymes to promote the growth of plants, the minicell described herein can deliver other high-value biomolecules that play a role in promoting the growth of plants. These biomolecules include, but are not 23 303137700 limited to plant hormones, such as the auxin IAA, peptides, primary metabolites, and secondary metabolites. [00100] In some embodiments, the biologically active compounds are pheromones to improve and modify chemical reactions to help the plants grow and fight stresses as biostimulants. [00101] In other embodiments, the delivery of biocontrols and biostimulants can be assisted through binding domains expressed on a surface of minicells. For example, minicells can express a binding domain such as a carbohydrate binding module (CBM) to be targeted to plants. These domains allow for better retention on plant surfaces, preventing runoff or drift. In some embodiments, minicells express a fusion protein comprising at least one surface expressing moiety and at least one target cell adhesion moiety, wherein said target cell adhesion moiety comprises a carbohydrate binding module. The target cell adhesion moiety comprises a carbohydrate binding module selected from the group consisting of: a cellulose binding domain, a xylan binding domain, a chitin binding domain, and a lignin binding domain. [00102] In other embodiments, minicells can also express various proteins that encourage them to be uptaken by plants for invasive delivery through the leaf surface or roots. In some embodiments, minicells can express and display biologically active compound such as polypeptide and/or proteins on their surface. In other embodiments, minicells can express and display both surface expressed binding proteins and biologically active compound such as polypeptide and/or proteins on their surface. [00103] The surface expressed binding proteins are as a carbohydrate binding module (CBM) described above. The biologically/enzymatically active polypeptide/proteins, which are surface-expressed, comprise cell stimulation moiety and/or cell degradation moiety. Non- limiting examples of such active proteins include, but are not limited to, ACC-deaminase, chitinase, cellulase, phytase, chitinase, protease, phosphatase, nucleases, lipases, glucanases, xylanases, amylases, peptidases, peroxidases, ligninases, pectinases, hemicellulases, and keratinases. [00104] In some embodiments, these proteins are expressed exogenously and encapsulated into the minicells. In other embodiments, these proteins are internally expressed and immobilized on the surface of the minicells. The biologically active compounds such as such proteins are either encapsulated within the minicells after being expressed outside of the minicells or internally expressed within the minicells and displayed on the surface of the minicells. In further embodiments, the minicells express at least one biologically active compound on its surface and encapsulate another biologically active compound at the same time. So, the minicell can carry at least two biologically active compounds within the minicells and on the 24 303137700 surface of the minicells. Non-limiting examples of such proteins include, but are not limited to ACC-deaminase, cellulase, phytase, chitinase, protease, phosphatase, nucleases, lipases, glucanases, xylanases, amylases, peptidases, peroxidases, ligninases, pectinases, hemicellulases, and keratinases. [00105] In some embodiments, the protein is lipase used as a biocontrol compound. In other embodiments, the protein is lipase used as a biostimulant compound. In further embodiments, the protein is ACC deaminase used as a biostimulant compound. In some embodiments, the protein is lipase used as a biocontrol compound. In other embodiments, the protein is lipase used as a biostimulant compound. In further embodiments, the protein is ACC deaminase used as a biostimulant compound. [00106] In some embodiments, minicells express a fusion protein comprising at least one surface expressing moiety and at least one target cell degradation moiety, wherein said target cell degradation moiety comprises an cutinase and cellulose. [00107] The present disclosure teaches production and encapsulation of the RNA biomolecule including antisense nucleic acid, dsRNA, shRNA, siRNA, miRNA, ribozyme, or aptamer during the fermentation cycle by utilizing the microorganism’s RNA synthesis and asymmetric division capabilities. This anucleated cell-based platform (i.e., minicell) and/or an agricultural composition addresses three critical issues that have posed a great challenge to the delivery of ribonucleic acid (RNA) to a system: (1) the scalable synthesis and encapsulation of RNA (2) the synthesized/encapsulated oligonucleotide payload must survive the process; (3) the targeted delivery of this RNA biomolecule such that it reaches the tissue or cells of interest and invokes the desired phenotypic response. Current forms of RNA delivery are direct coupling of siRNA to N-acetylgalactosamine (GalNAc), formulating the RNA (often chemically modified) with cationic lipids and other excipients protects the oligonucleotide from the environment to compact its size, making chemical modifications to stabilize oligonucleotides for RNAi DSSOLFDWLRQV^VXFK^DV^UHSODFLQJ^WKH^^ƍ-K\GUR[\O^JURXS^RQ^WKH^ULERVH^ULQJ^ZLWK^^ƍ-PHWKR[\^DQG^^ƍ- fluoro moieties. For dsRNA production, in vitro transcription is incredibly expensive compared to in vivo bacterial production of dsRNA. There are also Cell-Free and protein capsid processes for the production of dsRNA. The bacterial model is accompanied with the risk of environmental contamination due to proliferation of the modified species. This proliferation can have adverse and unforeseen consequences on the naturally existing species in the environment. Minicells result from naturally occurring mutations. The use of minicells for the purification and delivery of RNA allow for use the benefits of fermentation to scale the dsRNA production, without the risks associated with using genetically-modified bacteria. The use of 25 303137700 minicells is also better for the delivery of protoxins and enzymes than using genetically- modified bacteria as biopesticides. Overview [00108] RNA interference (RNAi) technology are used to specifically target pests, weeds, and pathogens for field control without the unwanted off-target effects of chemical pesticides. Due to their specificity, RNAi can be used to be added to other pesticides for maximum field control via the Integrated Pest Management (IPM) system, which is an effective and environmentally sensitive approach to pest management that relies on a combination of common-sense practices. [00109] In one aspect, the RNAi target can be designed to restore one or more pests’ susceptibility to chemical pesticides. Chemical pesticides work by targeting certain biological or metabolic pathways and specific receptors in those pathways of pests, pathogens, and weeds. Once chemical pesticides are continuously applied to pests, pathogens, and weeds, they develop resistance by mutating the receptors that are targeted by chemical pesticides. With the pesticide resistance developed, the pesticides lose efficacy for controlling pests, thereby leading to an increased dose of the pesticides required to achieve the expected level of control. If the pesticide dose increases however, it enters into a vicious cycle only leading to more resistance issues. [00110] In another aspect, pesticide resistance can be acquired by pests, pathogens, and weeds that evolve to express various biological or metabolic pathways that allow them to rapidly degrade the chemical pesticides or weaken mode of pesticide action. This natural evolution in response to pesticide application leads to another vicious cycle which leads to more resistance issues. [00111] Most, if not all, resistance issues are conventionally dealt with via Integrated Pest Management (IPM) to indirectly reduce resistance by providing rotational partners from different chemical classes. [00112] The present disclosure provides a novel approach to directly deal with pesticide resistance issues by combating the genetic evolution of pests, pathogens, and weeds and to prevent them from developing resistance. Examples of the disclosure include, but are not limited to, (i) minicells co-encapsulated RNAs (such as dsRNA, siRNA, miRNA, and antisense RNA) and chemical pesticides, (ii) minicell encapsulated RNAs paired with unencapsulated chemical pesticides, and (iii) minicell encapsulated RNAs paired with minicell encapsulated pesticides. 26 303137700 [00113] The present disclosure provides that this novel approach using a minicell platform for delivering a nucleic acid capable of inducing RNAi to a target along with chemical pesticides. [00114] The present disclosure also provides that this novel approach using a minicell platform for delivering a nucleic acid capable of inducing RNAi to a target along with biopesticides such as protoxins. [00115] The present disclosure provides that this novel approach using a minicell platform for delivering a nucleic acid capable of inducing RNAi to a target along with biopesticides and chemical pesticides. Minicells [00116] Minicells are the result of aberrant, asymmetric cell division, and contain membranes, peptidoglycan, ribosomes, RNA, protein, and often plasmids but no chromosome. (Frazer AC and Curtiss III, Production, Properties and Utility of Bacterial Minicells, Curr. Top. Microbial. Immunol. 69:1-84 (1975)). Because minicells lack chromosomal DNA, minicells cannot divide or grow, but they can continue other cellular processes, such as ATP synthesis, replication and transcription of plasmid DNA, and translation of mRNA. Although chromosomes do not segregate into minicells, extrachromosomal and/or episomal genetic expression elements may segregate, or may be introduced into minicells after segregation from parent cells. [00117] In embodiments, the minicells described herein are non-naturally occurring. [00118] In some embodiments, the disclosure provides a composition comprising a plurality of minicells. In some embodiments, the disclosure provides a composition comprising a plurality of minicells comprising at least one biologically active compound within said cell. In some embodiments, the disclosure provides a composition comprising a plurality of minicells, wherein each minicell of said plurality comprises an enzymatically active polypeptide displayed on the surface of the minicell, wherein said enzymatically active polypeptide has enzymatic activity. The enzymatic activity is derived from enzymatically active polypeptides disclosed in the present disclosure. [00119] In some embodiments, the invention provides a composition comprising a plurality of intact, bacterially-derived minicells. In some embodiments, the disclosure provides a composition comprising a plurality of intact, bacterially-derived minicells comprising at least one biologically active compound within said cell. In some embodiments, the invention provides a composition comprising a plurality of intact, bacterially-derived minicells, wherein each minicell of said plurality comprises an enzymatically active polypeptide displayed on the 27 303137700 surface of the bacterial minicell, wherein said enzymatically active polypeptide has enzymatic activity. In some embodiments, the composition comprises minicells which further comprise a second polypeptide displayed on the surface of the bacterial minicell, to increase adhesion to a subject and/or subjects including, but are not limited to substrates of enzymes, receptors, metal, plastic, soil, bacteria, fungi, pathogens, germs, plants, animals, human, and the like. In some embodiments, the composition comprises a mixture of minicells, wherein certain minicells within the mixed minicell population display the enzymatically active polypeptide or display the second polypeptide including subject adhesion increasing polypeptide or display both. (i) Eubacterial Minicells [00120] One type of minicell is a eubacterial minicell. For reviews of eubacterial cell cycle and division processes, see Rothfield et al., Annu. Rev. Genet., 33:423-48, 1999; Jacobs et al., Proc. Natl. Acad. Sci. USA, 96:5891-5893, May, 1999; Koch, Appl. and Envir. Microb., Vol. 66, No. 9, pp. 3657-3663; Bouche and Pichoff, Mol Microbiol, 1998. 29: 19-26; Khachatourians et al., J Bacteriol, 1973. 116: 226-229; Cooper, Res Microbiol, 1990.141: 17- 29; and Danachie and Robinson, “Cell Division: Parameter Values and the Process,” in: Escherichia Coli and Salmonella Typhimurium: Cellular and Molecular Biology, Neidhardt, Frederick C., Editor in Chief, American Society for Microbiology, Washington, D.C., 1987, Volume 2, pages 1578-1592, and references cited therein; and Lutkenhaus et al., “Cell Division,” Chapter 101 in: Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology, 2^nd Ed., Neidhardt, Frederick C., Editor in Chief, American Society for Microbiology, Washington, D.C., 1996, Volume 2, pages 1615-1626, and references cited therein. When DNA replication and/or chromosomal partitioning is altered, membrane- bounded vesicles “pinch off” from parent cells before transfer of chromosomal DNA is completed. As a result of this type of dysfunctional division, minicells are produced which contain an intact outer membrane, inner membrane, cell wall, and all of the cytoplasm components but do not contain chromosomal DNA. [00121] In some embodiments, the bacterially-derived minicells are produced from a strain, including, but are not limited to a strain of Escherichia coli, Bacillus spp., Salmonella spp., Listeria spp., Mycobacterium spp., Shigella spp., or Yersinia spp. In some embodiments, the bacterially-derived minicells are produced from a strain that naturally produces minicells. Such natural minicell producing strains produce minicells, for example, at a 2: 1 ratio (2 bacterial cells for every one minicell). In certain embodiments, exemplary bacterial strains that naturally produce minicells include, but are not limited to E. coli strain number P678-54, Coli Genetic Stock Center (CGSC) number: 4928 and B. subtilis strain CU403. 28 303137700 [00122] As one example, mutations in B. subtilis smc genes result in the production of minicells (Britton et al., 1998, Genes and Dev. 12:1254-1259; Moriya et al., 1998, Mol Microbiol 29:179-87). Disruption of smc genes in various cells is predicted to result in minicell production therefrom. [00123] As another example, mutations in the divIVA gene of Bacillus subtilis results in minicell production. When expressed in E. coli, B. subtilis or yeast Schizosaccharomyces pombe, a DivIVA-GFP protein is targeted to cell division sites therein, even though clear homologs of DivIVA do not seem to exist in E. coli, B. subtilis or S. pombe (David et al., 2000, EMBO J.19:2719-2727. Over- or under-expression of B. subtilis DivIVA or a homolog thereof may be used to reduce minicell production in a variety of cells. [00124] In some embodiments, the minicell-producing bacteria is a Gram-negative bacteria. The Gram-negative bacteria includes, but is not limited to, Escherichia coli, Salmonella spp. including Salmonella typhimurium, Shigella spp. including Shigella flexneri, Pseudomonas aeruginosa, Agrobacterium, Campylobacter jejuni, Lactobacillus spp., Neisseria gonorrhoeae, and Legionella pneumophila,. In some embodiments, the minicell-producing gram-negative bacteria can produce minicells naturally caused by endogenous or exogenous mutation(s) associated with cell division and/or chromosomal partitioning. In some embodiments, the minicell-producing bacteria comprises endogenous or exogenous gene(s) that is involved in cell division and/or chromosomal partitioning, where the gene is genetically modified such as by homologous recombination, compared to a corresponding wild-type gene. In some embodiments, the minicell-producing gram-negative bacteria is deficient in protease and/or its activity naturally and/or by genetic engineering techniques disclosed herein. In some embodiments, the protease-deficient minicell-producing gram-negative bacteria comprises a recombinant expression vector comprising a gene or genes that is involved in a protein of interest disclosed in the present disclosure. [00125] In some embodiments, the minicell-producing bacteria can be a Gram-positive bacteria. The Gram-positive bacteria includes, but is not limited to, Bacillus subtilis, Bacillus cereus, Corynebacterium Glutamicum, Lactobacillus acidophilus, Staphylococcus spp., or Streptococcus spp. In some embodiments, the minicell-producing gram-positive bacteria can produce minicells naturally caused by endogenous or exogenous mutation(s) associated with cell division and/or chromosomal partitioning. In some embodiments, the minicell-producing gram-positive bacteria comprises endogenous or exogenous gene(s) that is involved in cell division and/or chromosomal partitioning, where the gene is genetically modified such as by homologous recombination, compared to a corresponding wild-type gene. In some 29 303137700 embodiments, the minicell-producing gram-positive bacteria is deficient in protease and/or its activity naturally and/or by genetic engineering techniques disclosed herein. In some embodiments, the protease-deficient minicell-producing gram-positive bacteria comprises a recombinant expression vector comprising a gene or genes that is involved in a protein of interest disclosed in the present disclosure. [00126] The minicell-producing bacteria can be a Extremophilic bacteria. The Extremophilic bacteria includes, but is not limited to, Thermophiles including Thermus aquaticus, Psychrophiles, Piezophiles, Halophilic bacteria, Acidophile, Alkaliphile, Anaerobe, Lithoautotroph, Oligotroph, Metallotolerant, Oligotroph, Xerophil or Polyextremophile. In some embodiments, the minicell-producing Extremophilic bacteria can produce minicells naturally caused by endogenous or exogenous mutation(s) associated with cell division and/or chromosomal partitioning. In some embodiments, the minicell-producing Extremophilic bacteria comprises endogenous or exogenous gene(s) that is involved in cell division and/or chromosomal partitioning, where the gene is genetically modified such as by homologous recombination, compared to a corresponding wild-type gene. In some embodiments, the minicell-producing Extremophilic bacteria is deficient in protease and/or its activity naturally and/or by genetic engineering techniques disclosed herein. In some embodiments, the protease- deficient minicell-producing Extremophilic bacteria comprises a recombinant expression vector comprising a gene or genes that is involved in a protein of interest disclosed in the present disclosure. (ii) Eukaryotic Minicells [00127] Achromosomal eukaryotic minicells (i.e., anucleate cells) are within the scope of the disclosure. Yeast cells are used to generate fungal minicells. See, e.g., Lee et al., Ibd1p, a possible spindle pole body associated protein, regulates nuclear division and bud separation in Saccharomyces cerevisiae, Biochim Biophys Acta 3:239-253, 1999; Kopecka et al., A method of isolating anucleate yeast protoplasts unable to synthesize the glucan fibrillar component of the wall J Gen Microbiol 81:111-120, 1974; and Yoo et al., Fission yeast Hrp1, a chromodomain ATPase, is required for proper chromosome segregation and its overexpression interferes with chromatin condensation, Nucl Acids Res 28:2004-2011, 2000. Cell division in yeast is reviewed by Gould and Simanis, The control of septum formation in fission yeast, Genes & Dev 11:2939-51, 1997). [00128] In some embodiments, the eukaryotic minicells can be produced from yeast cells, such as Saccharomyces cerevisiae, Pichia pastoris and/or Schizosaccharomyces pombe. 30 303137700 [00129] As one example, mutations in the yeast genes encoding TRF topoisomerases result in the production of minicells, and a human homolog of yeast TRF genes has been stated to exist (Castano et al., 1996, Nucleic Acids Res 24:2404-10). Mutations in a yeast chromodomain ATPase, Hrp1, result in abnormal chromosomal segregation; (Yoo et al., 2000 Nuc. Acids Res. 28:2004-2011). Disruption of TRF and/or Hrp1 function is predicted to cause minicell production in various cells. Genes involved in septum formation in fission yeast (see, e.g., Gould et al., 1997 Genes and Dev. 11:2939-2951) can be used in like fashion. [00130] Platelets are a non-limiting example of eukaryotic minicells. Platelets are anucleate cells with little or no capacity for de novo protein synthesis. The tight regulation of protein synthesis in platelets (Smith et al., 1999, Vasc Med 4:165-72) may allow for the over- production of exogenous proteins and, at the same time, under-production of endogenous proteins. Thrombin-activated expression elements such as those that are associated with Bcl-3 (Weyrich et al., Signal-dependent translation of a regulatory protein, Bcl-3, in activated human platelets, Cell Biology 95:5556-5561, 1998) may be used to modulate the expression of exogenous genes in platelets. [00131] As another non-limiting example, eukaryotic minicells are generated from tumor cell lines (Gyongyossy-Issa and Khachatourians, Tumour minicells: single, large vesicles released from cultured mastocytoma cells (1985) Tissue Cell 17:801-809; Melton, Cell fusion-induced mouse neuroblastomas HPRT revertants with variant enzyme and elevated HPRT protein levels (1981) Somatic Cell Genet. 7: 331-344). [00132] Yeast cells are used to generate fungal minicells. See, e.g., Lee et al., Ibd1p, a possible spindle pole body associated protein, regulates nuclear division and bud separation in Saccharomyces cerevisiae, Biochim Biophys Acta 3:239-253, 1999; Kopecka et al., A method of isolating anucleate yeast protoplasts unable to synthesize the glucan fibrillar component of the wall J Gen Microbiol 81:111-120, 1974; and Yoo et al., Fission yeast Hrp1, a chromodomain ATPase, is required for proper chromosome segregation and its overexpression interferes with chromatin condensation, Nucl Acids Res 28:2004-2011, 2000. Cell division in yeast is reviewed by Gould and Simanis, The control of septum formation in fission yeast, Genes & Dev 11:2939-51, 1997). In some embodiments, the present disclosure teaches production of yeast minicells (iii) Archaebacterial Minicells [00133] The term “archaebacterium” is defined as is used in the art and includes extreme thermophiles and other Archaea (Woese, C.R., L. Magrum. G. Fox. 1978. Archaebacteria. Journal of Molecular Evolution. 11:245-252). Three types of Archaebacteria are halophiles, 31 303137700 thermophiles and methanogens. By physiological definition, the Archaea (informally, archaes) are single-cell extreme thermophiles (including thermoacidophiles), sulfate reducers, methanogens, and extreme halophiles. The thermophilic members of the Archaea include the most thermophilic organisms cultivated in the laboratory. The aerobic thermophiles are also acidophilic; they oxidize sulfur in their environment to sulfuric acid. The extreme halophiles are aerobic or microaerophilic and include the most salt tolerant organisms known. The sulfate- reducing Archaea reduce sulfate to sulfide in extreme environment. Methanogens are strict anaerobes, yet they gave rise to at least two separate aerobic groups: the halophiles and a thermoacidophilic lineage. Non-limiting examples of halophiles include Halobacterium cutirubrum and Halogerax mediterranei. Non-limiting examples of methanogens include Methanococcus voltae; Methanococcus vanniela; Methanobacterium thermoautotrophicum; Methanococcus voltae; Methanothermus fervidus; and Methanosarcina barkeri. Non-limiting examples of thermophiles include Azotobacter vinelandii; Thermoplasma acidophilum; Pyrococcus horikoshii; Pyrococcus furiosus; and Crenarchaeota (extremely thermophilic archaebacteria) species such as Sulfolobus solfataricus and Sulfolobus acidocaldarius. [00134] Archaebacterial minicells are within the scope of the invention. Archaebacteria have homologs of eubacterial minicell genes and proteins, such as the MinD polypeptide from Pyrococcus furiosus (Hayashi et al., EMBO J. 20:1819-28, 2001). It is thus possible to create Archaebacterial minicells by methods such as, by way of non-limiting example, overexpressing the product of a min gene isolated from a prokaryote or an archaebacterium; or by disrupting expression of a min gene in an archaebacterium of interest by, e.g., the introduction of mutations thereof or antisense molecules thereto. See, e.g., Laurence et al., Genetics 152:1315-1323, 1999. [00135] By physiological definition, the Archaea (informally, archaes) are single-cell extreme thermophiles (including thermoacidophiles), sulfate reducers, methanogens, and extreme halophiles. The thermophilic members of the Archaea include the most thermophilic organisms cultivated in the laboratory. The aerobic thermophiles are also acidophilic; they oxidize sulfur in their environment to sulfuric acid. The extreme halophiles are aerobic or microaerophilic and include the most salt tolerant organisms known. The sulfate-reducing Archaea reduce sulfate to sulfide in extreme environment. Methanogens are strict anaerobes, yet they gave rise to at least two separate aerobic groups: the halophiles and a thermoacidophilic lineage. In some embodiments, the present disclosure teaches production of archaeal minicells. 32 303137700 (iv) Minicells derived from endophytes [00136] An endophyte is an endosymbiont, often a bacterium or fungus, that lives within a plant for at least part of its life cycle. The endophyte can transport itself from the environment to internal organs of plants. Non-limiting examples of endophytes include Acidovorax facilis, Bradyrhizobium, Rhizobium, Rhodococcus rhodochrous, Colletotrichum, Curvularia, Epichloë, Fusarium, Mycosphaerella, Neotyphodium, Piriformospora, and Serendipita. In some embodiments, the present disclosure teaches production of endophyte-derived minicells. In other embodiments, endophyte-derived minicells can enter into internal plant cell, tissues, or organs, and function as an invasive minicell. [00137] Fungal endophytes have the ability to colonize inter- or intra-cellularly. The colonization process involves several steps, including host recognition, spore germination, penetration of the epidermis and tissue multiplication. Once the endophytes are successfully colonized in the host tissue, the endophytic niche becomes established. In the endophytic niche, endophytes will obtain a reliable source of nutrition from the plant fragment, exudates and leachates and protect the host against other microorganisms (Gao et al., 2010). In some embodiments, minicells produced from fungal endophytes can transmit the active compounds within and/or on their surface to a target using their invasive capability (v) Minicells derived from plant pathogen bacteria [00138] The present disclosure provides plant pathogen bacteria, which can be utilized for minicell production, including but are not limited to (1) Pseudomonas syringae pathovars; (2) Ralstonia solanacearum; (3) Agrobacterium tumefaciens; (4) Xanthomonas oryzae pv. oryzae; (5) Xanthomonas campestrispathovars; (6) Xanthomonas axonopodis pathovars; (7) Erwinia amylovora; (8) Xylella fastidiosa; (9) Dickeya (dadantii and solani); (10) Pectobacterium carotovorum (and Pectobacterium atrosepticum), (11) Clavibacter michiganensis (michiganensis and sepedonicus), (12) Pseudomonas savastanoi, and (13) Candidatus Liberibacter asiaticus. Such plant pathogen bacteria natively have the capacity to penetrate and invade into internal host tissues in their natural state. In some embodiments, minicells derived from plant pathogen bacteria described above can naturally deliver biologically active compounds disclosed herein into internal cells, tissues, and/or organs of a target host in their natural ability of invasion, penetration, and/or transmission into internal parts of a target. [00139] From example, some pathogen bacteria are found to secrete cell wall-degrading endoglucanase and endopolygalacturonase, potentially explaining penetration into the root endosphere. Other pathogen bacteria can penetrate through the stomata into the substomatal 33 303137700 chamber, and colonization of the intercellular spaces of the leaf mesophyll. The minicells produced from these pathogen bacteria possess and utilize natural ability of invading, penetrating and/or transmitting for scalable and targeted delivery of active compounds disclosed herein. Bacterial Minicell Production [00140] Minicells are produced by parent cells having a mutation in, and/or overexpressing, or under expressing a gene involved in cell division and/or chromosomal partitioning, or from parent cells that have been exposed to certain conditions, that result in aberrant fission of bacterial cells and/or partitioning in abnormal chromosomal segregation during cellular fission (division). The term “parent cells” or “parental cells” refers to the cells from which minicells are produced. Minicells, most of which lack chromosomal DNA (Mulder et al., Mol Gen Genet, 221: 87-93, 1990), are generally, but need not be, smaller than their parent cells. [00141] Minicells are achromosomal, membrane-encapsulated biological nanoparticles (أ400 nm) that are formed by bacteria following a disruption in the normal division apparatus of bacterial cells. Minicells can also be 400nm to 650nm in size. In essence, minicells are small, metabolically active replicas of normal bacterial cells with the exception that they contain no chromosomal DNA and as such, are non-dividing and non-viable. Although minicells do not contain chromosomal DNA, the ability of plasmids, RNA, native and/or recombinantly expressed proteins, and other metabolites have all been shown to segregate into minicells. Some methods of construction of minicell-producing bacterial strains are discussed in detail in U.S. patent application Ser. No.10/154,951(US Publication No. US/2003/0194798 A1), which is hereby incorporated by reference in its entirety. [00142] Disruptions in the coordination between chromosome replication and cell division lead to minicell formation from the polar region of most rod-shaped prokaryotes. Disruption of the coordination between chromosome replication and cell division can be facilitated through the overexpression of some of the genes involved in septum formation and binary fission. Alternatively, minicells can be produced in strains that harbor mutations in genes that modulate septum formation and binary fission. Impaired chromosome segregation mechanisms can also lead to minicell formation as has been shown in many different prokaryotes. (i) Plasmid Based Methods of Minicell Production [00143] In some embodiments, minicell production can be achieved by the overexpression or mutation of genes involved in the segregation of nascent chromosomes into daughter cells. For example, mutations in the parC or mukB loci of E. coli have been demonstrated to produce 34 303137700 minicells. The overexpression or mutation of a cell division gene capable of driving minicell production in one family member, can be used to produce minicells in another. For example, it has been shown that the overexpression E. coli ftsZ gene in other Enterobacteriacea family members such as Salmonella spp. and Shigella spp as well as other class members such as Pseudomonas spp. will result in similar levels of minicell production. [00144] In some embodiments, minicells can be produced in E. coli by the overproduction of the protein FtsZ which is an essential component of the Min division system by which E. coli operates. Overproduction of this protein in E. coli results in the inability for this ring to be spatially restricted to the midsection of the cell, thus resulting in production of minicells upon cell division. Because the overproduction of FtsZ can create minicells, it can be overexpressed using a plasmid based system. [00145] The same can be demonstrated in the mutation-based minicell producing bacterial strains. For example, deletion of the Min locus in any of bacterial strains results in minicell production. Cell division genes in which mutation can lead to minicell formation include but are not limited to the min genes (such as minC, minD, and minE). [00146] In some embodiments, E. coli rely on the min system in order to ensure proper replication of parent cells into daughter cells. This min system (known as the minB operon) consists of 3 parts, minD, minC, and minE. These genes work together in order to control the placement of the Z-ring which is comprised of polymerized FtsZ protein. MinC consists of two distinct domains, both of which interact directly with the FtsZ protein in order to inhibit polymerization (Z-ring formation). MinD is a protein that is associated with the membrane that forms at one of the cell’s poles and polymerizes toward the cell’s mid-point. It binds MinC which is distributed throughout the cytoplasm. MinE is a protein that binds to MinD as well and releases MinC. It polymerizes into a ring like shape and oscillates from pole to pole in the cell. [00147] In some embodiments, this system can be manipulated in order to shift the Z-ring to a polar end of the cell which excludes the nucleoid DNA upon completion of replication. The Z-ring can be shifted by not allowing the cell to sequester MinC to the polar ends of the cell. In the absence of MinC or MinD, or overexpression of MinE, E. coli cells will form achromosomal and/or anucleate cells. The FtsZ and the Min systems for causing asymmetrical cell division are exemplified by Piet et al, 1990, Proc. Natl. Acad. Sci. USA 87:1129-1133 and Xuan-Chuan et al, 2000, J. Bacteriol. 182(21):6203-62138, each of which is incorporated herein by reference. 35 303137700 [00148] Genes can be introduced in a site directed fashion using homologous recombination. Homologous recombination permits site specific modifications in endogenous genes and thus inherited or acquired mutations may be corrected, and/or novel alterations may be engineered into the genome. Homologous recombination and site-directed integration in plants are discussed in, for example, U.S. Patent Nos. 5,451,513; 5,501,967 and 5,527,695. [00149] In some embodiments, minicells are produced by deleting, mutating, knocking out, or disrupting minC, minD, and/or minC and minD gene(s) in bacteria by traditional gene engineering techniques including homologous recombination. In other embodiments, minicells are produced by overexpressing certain genes such as ftsZ and/or minE in bacteria. (ii) Controlled Production of Minicells [00150] In some embodiments, the present disclosure teaches mutating cell populations by introducing, deleting, or replacing selected portions of genomic DNA. Thus, in some embodiments, the present disclosure teaches methods for targeting mutations to a specific locus such as ftsZ, minC, minD, minC/D, and minE. In other embodiments, the present disclosure teaches the use of gene editing technologies such as ZFNs, TALENS, CRISPR or homing endonucleases, to selectively edit target DNA regions. In aspects, the targeted DNA regions is ftsZ, minC, minD, minC/D, and minE. [00151] Engineered nucleases such as zinc finger nucleases (ZFNs), Transcription Activator Like Effector Nucleases (TALENs), engineered homing endonucleases, and RNA or DNA guided endonucleases, such as CRISPR/Cas such as Cas9 or CPF1, are particularly appropriate to carry out some of the methods of the present disclosure. Additionally or alternatively, RNA targeting systems can use used, such as CRISPR/Cas systems have RNA targeting nucleases. [00152] In some embodiments, one skilled in the art can appreciate that the Cas9 disclosed herein can be any variant described in the literature, including but not limited to the functional mutations described in: Fonfara et al. Nucleic Acids Res.2014 Feb;42(4):2577-90; Nishimasu H. et al. Cell.2014 Feb 27; 156(5):935-49; Jinek M. et al. Science.2012337:816-21; and Jinek M. et al. Science.2014 Mar 14; 343(6176); see also U.S. Pat. App. No.13/842,859 filed March 15, 2013, which is hereby incorporated by reference; further, see U.S. Pat. Nos. 8,697,359; 8,771,945; 8,795,965; 8,865,406; 8,871,445; 8,889,356; 8,895,308; 8,906,616; 8,932,814; 8,945,839; 8,993,233; and 8,999,641, which are all hereby incorporated by reference. Thus, in some embodiments, the systems and methods disclosed herein can be used with the wild type Cas9 protein having double-stranded nuclease activity, Cas9 mutants that act as single stranded nickases, deactivated Cas9 (dCas9) that has no nuclease activity, or other mutants with modified nuclease activity. 36 303137700 [00153] In some examples, a Type II nuclease may be catalytically dead (e.g. dCas9, “dead Cas9,” “deactivated Cas9”) such that it binds to a target sequence, but does not cleave. dCAS9 is a variant of the CAS9 protein (CRISPR) that has had its active site altered to no longer be able to edit genomes, but can still bind to highly specific segments of the genome using a guide RNA. This protein can stop transcription of the gene if bound. In some embodiments, the dCAS9 gene can be placed under inducible control so that its expression would be controlled. The guide RNA corresponding to the knockout within the Min system could be included on a plasmid or cut into the genome and placed under inducible control. Upon induction with this system, the guide RNA would direct the dCAS9 protein to the gene within the Min system in order to stop its expression. The stopping of expression of this gene such as minC, minD, and minC/D would result in the formation of minicells. (iii)Antibiotic Resistance Knock in-Knock out [00154] In some embodiments, the present disclosure teaches uses of the genetic manipulation technique using Lambda-Red recombination system in order to edit genome integrated with exogenous expression cassette such as an selectable marker such as antibiotic resistant gene. In some embodiments, an selectable marker such as antibiotic resistant gene is integrated into the host genome (e.g. bacteria) in order to knockout minC/D/CD gene for inducing minicell production. If the marker with antibiotic resistance is no longer desired after successfully selecting the minicells in which the target gene (such as minC/D/CD) is knocked out, the flippase can be used to remove the integrated antibiotic resistant gene cassette from the host genome. A fragment of linear DNA is inserted into the genome directed by that fragment homology to the genome. This can be used to knock in genes of interest or to knockout genes of interest by replacing them with an antibiotic resistance cassette such as Chloramphenicol- resistant gene, kanamycin-resistant gene, spectinomycin-resistant gene, streptomycin-resistant gene, ampicillin-resistant gene, tetracycline-resistant gene, erythromycin-resistant gene, bleomycin-resistant gene, and bleomycin-resistant gene. A successful genetic manipulation is then selected for using this antibiotic resistance cassette. If a flippase recombination target (FRT) site is included within the resistance cassette for further genetic manipulations, it can be used for removing the antibiotic resistant gene integrated into the genome in vivo after selection of target minicells. The enzyme used for this is recombinase flippase and is often expressed from a plasmid that can be removed from the cell line using a temperature sensitive origin of replication. Recombinase flippase recognizes two identical FRT sites on both the 5’ and 3’ ends of the antibiotic resistance cassette and removes the DNA between the two sites. In some 37 303137700 embodiments, the FRT site can be included within an antibiotic resistance cassette to remove the antibiotic resistance cassette after its use. Strains for Minicell Production [00155] A E. coli P678-54 strain is obtained from Coli Genetic Stock Center (CGSC), and is used to produce minicells (Adler et al., 1967, Proc. Natl. Acad. Sci. USA 57:321-326; Inselburg J, 1970 J. Bacteriol.102(3):642-647; Frazer 1975, Curr. Topics Microbiol. Immunol.69:1-84). [00156] In some embodiments, a minicell is produced from a P678-54 E. coli parental strain. The minicell produced from P678-54 parental bacterial strain is used as an anucleated cell- based platform (i.e. minicell platform) and/or an agricultural composition for the encapsulation and delivery of biologically active compounds. (i) Protease-deficient bacterial strains [00157] The present disclosure provides the production of minicells from B strains using genetically-engineering techniques including B strains including BL21, BL21 (DE3), and BL21-AI are deficient in Lon protease (cytoplasm) and OmpT protease (outer membrane). Accordingly, B strains as protease-deficient strains can be utilized to create protease-deficient and/or protease-deficient minicells. The DE3 designation means that respective strains contain WKH^^'(^^O\VRJHQ^WKDW^FDUULHV^WKH^JHQH^IRU^7^^51$^SRO\PHUDVH^XQder control of the lacUV5 promoter. IPTG is required to maximally induce expression of the T7 RNA polymerase in order to express recombinant genes cloned downstream of a T7 promoter. BL21(DE3) is suitable for expression from a T7 or T7-lac promoter or promoters recognized by the E.coli RNA polymerase: e.g. lac, tac, trc, ParaBAD, PrhaBAD and also the T5 promoter. The genotype of BL21 (DE3) is: IKX$^^>ORQ@^RPS7^JDO^^^^'(^^^>GFP@^¨KVG6 ^^'(^^ ^^^V%DP+,R^¨(FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^L^^^¨QLQ^^ [00158] BL21-AI E. coli contains a chromosomal insertion of the gene encoding T7 RNA polymerase (RNAP) into the araB locus of the araBAD operon, placing regulation of T7 RNAP under the control of the arabinose-inducible araBAD promoter. Therefore, this strain is especially useful for the expression of genes that may be toxic to other BL21 strains where basal expression of T7 RNAP is leaky. The BL21-AI strain does not contain the Ion protease and is deficient in the outer membrane protease, OmpT. The genotype of BL21-AI is F- ompT hsdS_B (r_B- m_B-) gal dcm araB::T7RNAP-tetA. The BL21-AI has an arabinose promoter that controls the production T7 RNA Polymerase, while the BL21 (DE3) has a lac promoter that controls the production of the T7 RNA Polymerase. This is significant because the lac 38 303137700 promotion system is leaky. Therefore, the BL21-AI protein production is more tightly regulated due to the arabinose promotion system. [00159] The present disclosure teaches that LPS (Lipopolysaccharide) modified BL21 (DE3) cells can be used. The LPS of the E. Coli is modified to be significantly less toxic. This LPS modified BL21 (DE3) cells if necessary. This could also be branched out to other gram- negative bacterial cells. Safe usage of gram-negative cells can be beneficial for minicell and/or an agricultural composition. [00160] ClearColi® BL21(DE3) cells are the commercially available competent cells with a modified LPS (Lipid IVA) that does not trigger the endotoxic response in diverse cells. For example, ClearColi cells lack outer membrane agonists for hTLR4/MD-2 activation; therefore, activation of hTLR4/MD-2 signaling by ClearColi® is several orders of magnitude lower as compared with E. coli wild-type cells. Heterologous proteins prepared from ClearColi® are virtually free of endotoxic activity. After minimal purification from ClearColi cells, proteins or plasmids (which may contain Lipid IVA) can be used in most applications without eliciting an endotoxic response in human cells. In ClearColi cells, two of the secondary acyl chains of the normally hexa-acylated LPS have been deleted, eliminating a key determinant of endotoxicity in eukaryotic cells. The six acyl chains of the LPS are the trigger which is recognized by the Toll-like receptor 4 (TLR4) in complex with myeloid differentiation factor 2 (MD-2), causing activation of NF-ڡ%^ DQG^SURGXFWLRQ^RI^Sroinflammatory cytokines. The deletion of the two secondary acyl chains results in lipid IVA, which does not induce the formation of the activated heterotetrameric TLR4/MD-2 complex and thus does not trigger the endotoxic response. In ClearColi® BL21(DE3) Electrocompetent Cells 4 MA145 Rev. 31OCT2016 addition, the oligosaccharide chain is deleted, making it easier to remove the resulting lipid IVA from any downstream product. [00161] In some embodiments, protease-deficient minicells disclosed herein are produced from protease-deficient parental strains including, but are not limited to, BL21 (DE3), BL21- AI and LPS-modified BL21 (DE3). In other embodiments, BL21 (DE3), BL21-AI and LPS- modified BL21 (DE3) strains are genetically engineered by deleting, mutating, knocking out, or disrupting minC, minD, and/or minC and minD gene(s) to induce minicell production. In other embodiments, BL21 (DE3), BL21-AI and LPS-modified BL21 (DE3) strains are genetically engineered by overexpressing ftsZ and/or minE genes to induce minicell production. [00162] In further embodiments, the present disclosure provides a new minicell-producing strain named as B8. This strain is the protease-deficient minicell-producing strain without the 39 303137700 T7 RNA Polymerase. This minicell strain is produced from the BL21 (DE3) strain. While knocking out minC/D/CD, the T7 RNA Polymerase was silenced due to the homology of the introduced knockout via Lambda Red Transformation. This strain can be used for a need of a protease-deficient minicell, but not having the T7 RNA Polymerase. In some embodiments, minicells displayed an enzymatically active polypeptide such as complicated or toxic proteins on their surface, need to be more controlled and slower expression of the desired but complicated or toxic proteins. [00163] The present disclosure teaches genotypes of newly-generated protease-deficient minicell strains comprising i) minC-deleted BL21(DE3); IKX$^^>ORQ@^RPS7^JDO^^^^'(^^^>GFP@^ ¨KVG6 ^^'(^^ ^^^V%DP+,R^¨(FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^L^^^¨QLQ^^¨PLQ&^^ ii) minD-deleted BL21(DE3); IKX$^^>ORQ@^RPS7^JDO^^^^'(^^^>GFP@^¨KVG6 ^^'(^^ ^^^V%DP+,R^ ¨(FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^L^^^¨ QLQ^^¨ PLQ'^^LLL^ minC/D-deleted BL21(DE3); IKX$^^ >ORQ@^ RPS7^ JDO^ ^^^ '(^^^ >GFP@^ ¨KVG6 ^^ '(^^ ^ ^^ V%DP+,R^ ¨(FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^ L^^^ ¨QLQ^^ ¨PLQ&^ ¨PLQ'^^ iv) minC-deleted BL21-AI; F- ompT hsdS_B (r_B- m_B-) gal dcm araB::T7RNAP-tetA ¨PLQ&^^ v) minD-deleted BL21-AI; F- ompT hsdS_B (r_B- m_B-) gal dcm araB::T7RNAP-tetA ¨PLQ'^^YL^ minC/D-deleted BL21-AI; F- ompT hsdS_B (r_B- m_B-) gal dcm araB::T7RNAP-tetA ¨PLQ&^¨PLQ'^^YLL^ minC-deleted LPS- modified BL21(DE3); PVE$^^^^¨JXW4^¨NGV'^¨OS[/^¨OS[0^¨SDJ3^¨OS[3^¨HSW$^¨minC, viii) minD-deleted LPS-modified BL21(DE3); PVE$^^^^¨JXW4^¨NGV'^¨OS[/^¨OS[0^¨SDJ3^¨OS[3^ ¨HSW$^¨minD, ix) minC/D-deleted LPS-modified BL21(DE3); PVE$^^^^¨JXW4^¨NGV'^¨OS[/^ ¨OS[0^¨SDJ3^¨OS[3^¨HSW$^¨minC, ¨minD, x) minC-deleted B8 with suppression on T7 RNA polymerase activity; IKX$^^>ORQ@^RPS7^JDO^^^^'(^^^>GFP@^¨ KVG6 ^^'(^^ ^^^V%DP+,R^¨ (FR5,- B int::(lacI::PlacUV5::T7 JHQH^^^L^^^¨QLQ^^¨PLQ&^^[L^^minD-deleted B8 with suppression on T7 RNA polymerase activity; IKX$^^>ORQ@^RPS7^JDO^^^^'(^^^>GFP@^¨KVG6 ^^'(^^ ^^^V%DP+,R^ ¨(FR5,-B int::(lacI::PlacUV5::T7 gene1^^L^^^¨ QLQ^^¨ PLQ'^^DQG^[LL^^minC/D-deleted B8 with suppression on T7 RNA polymerase activity; fhuA2 [lon] ompT JDO^^^^'(^^^>GFP@^¨KVG6 ^^ '(^^ ^^^V%DP+,R^¨(FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^L^^^¨QLQ^^¨PLQ&^¨PLQ'^ [00164] Minicells that have segregated from parent cells lack chromosomal and/or nuclear components, but retain the cytoplasm and its contents, including the cellular machinery required for protein expression. In some embodiments, minicells are protease-deficient because the parent cells are protease-deficient strains. Although chromosomes do not segregate into minicells, extrachromosomal and/or episomal genetic expression elements may segregate, or may be introduced into minicells after segregation from parent cells. In some embodiments, the disclosure is drawn to protease-deficient minicells comprising an expression element, 40 303137700 which may be an inducible expression element. The inducible expression element such as an inducible promoter can be introduced to a recombinant plasmid used for homologous recombination to knock out and/or delete gene(s) involved to cell division and/or chromosomal partitioning such as minC, minD, and minC/D, a recombinant expression vector to overexpress gene(s) involved to cell division and/or chromosomal partitioning such as ftsZ and minE, and a recombinant expression vector for expressing an enzymatically active polypeptide including a protein of interest disclosed herein. In further embodiments, the inducible expression element comprises expression sequences operably linked to an open reading frame (ORF) that encodes proteins of interest disclosed herein. Optionally, at any point in the method, an inducing agent is provided in order to induce expression of an ORF that encodes proteins of interest disclosed herein. [00165] In some embodiments, the disclosure teaches methods of making a protease-deficient bacterial minicell comprising a recombinant fusion protein that is not naturally found in parental cells. In some embodiment, the disclosure teaches method of preparing protease- deficient minicells from the host cells. [00166] In other embodiments, the present disclosure teaches production of protease-deficient minicells from B. subtilis strains such as CU403 DIVIVA, CU403,DIVIVB,SPO-, CU403,DIVIVB and CU403,DIVIVB1 using by deleting, mutating, knocking out, or disrupting gene encoding WprA protease. [00167] B. subtilis genetic manipulations work slightly differently than genetic manipulations in E. coli. B. subtilis is known to readily undergo homologous recombination if DNA containing homology to the existing genome is inserted. This is unlike E. coli; E. coli has mechanisms in place to degrade any non-natural linear DNA present. This difference can be utilized in order to knockout genes by designing an antibiotic resistance cassette flanked by homologous arms which correspond to the start and end of the gene that is desired to be knockout out. [00168] The present disclosure provides the production of minicells from B. subtilis using genetically-engineering techniques. In some embodiments, B. subtilis strains including, but are not limited to CU403 DIVIVA (BGSC No. 1A196), CU403,DIVIVB,SPO- (BGSC No. 1A197), CU403,DIVIVB (BGSC No. 1A292), CU403,DIVIVB1 (BGSC No. 1A513), KO7 can be used as parental bacterial cells to produce minicells. B. subtilis strains are the commercially available and can be obtained from Bacillus Genetic Stock Center (BGSC). The catalog of strains is available on the document provided by publicly accessible BGSC webpage (www.bgsc.org/_catalogs/Catpart1.pdf). 41 303137700 [00169] In some embodiments, Bacillus Subtilis stains including, but are not limited to CU403 DIVIVA, CU403,DIVIVB,SPO-, CU403,DIVIVB and CU403,DIVIVB1 can be genetically modified by knocking out gene encoding WprA Protease in these strains. WprA protease is known as one of the harshest proteases. [00170] In order to knock out, delete, and or remove WprA-encoding gene from B. subtilis strains, the pUC18 WprA-CamR vector is used. This vector has the homologous arms corresponding to the gene coding for WprA cell wall protease that naturally occurs in B. subtilis which is undesirable for protein surface expression. These homologous arms flank a chloramphenicol resistance cassette in order to allow for selection. After the homologous recombination via the homologous arms within the host cells, the WprA-encoding nucleotide except the homologous arm is replaced with the chloramphenicol selection marker gene. This plasmid can replicate within E. coli due to its origin of replication, thus when transformed into B. subtilis it cannot replicate. After transformation, colonies are selected for using chloramphenicol in order to isolate the colonies in which the knockout of WprA successfully occurs. Because the plasmid cannot replicate in B. subtilis, only the cells can survive against the presence of chloramphenicol if the recombinant cassette having the chloramphenicol resistant marker gene is integrated to the genome of the B. subtilis cell by homologous recombination. [00171] B. subtilis secretes no fewer than seven proteases during vegetative growth and stationary phase. Strains in which multiple protease genes have been inactivated have proved to be superior to wild type strains in production of foreign proteins. The KO7 is prototrophic, free of secreted proteases, and have marker-free deletions in PY79 genetic background. This KO7 is available from the BGSC as accession number 1A1133. KO7 Genotype: ǻQSU( ǻDSU( ǻHSU ǻPSU ǻQSU% ǻYSU ǻESU^ [00172] In some embodiments, a seven-protease deletion strain, B. subtilis KO7, can be used for B. subtilis minicell production by knocking out DIV-IVA and DIV-IVB using genetic engineering techniques described in the present disclosure. [00173] In some embodiments, a minicell is produced from a P678-54 E. coli wild strain. In other embodiments, a minicell is produced from a protease-deficient E. coli strain including BL21, BL21(DE3), BL21-AI, LPS-modified BL21 (DE3) and B8. In some embodiments, a minicell is produced from a parental bacterial cell deficient in WprA protease. In some embodiments, a minicell is produced from a protease deficient B. subtilis parental bacterial cell. In some embodiments, a minicell is produced from produced from a protease deficient KO7 B. subtilis parental bacterial cell. In other embodiments, a minicell is produced from a 42 303137700 protease deficient B. subtilis parental bacterial cell selected from the group consisting of: (1) CU403,DIVIVA; (2) CU403,DIVIVB,SPO-; (3) CU403,DIVIVB; and (4) CU403,DIVIVB1, wherein at least one protease encoding gene has been repressed, deleted, or silenced. In further embodiments, a minicell is produced from an eukaryotic cell. In further embodiments, the minicell produced as described above is used as an anucleated cell-based platform and/or an agricultural composition for the encapsulation and delivery of biologically active compounds. [00174] In some embodiments, minicells taught in the present disclosure is protease deficient or ribonuclease deficient. In some embodiments, said minicell is protease deficient. In some embodiments, said minicell is ribonuclease deficient. In some embodiments, said minicell is protease deficient and ribonuclease deficient. (ii) Ribonuclease-deficient bacterial strains [00175] The present disclosure provides the production of minicells from HT115 (DE3) using genetically-engineering techniques. HT115 (DE3) is a RNAi Feeding strain, which is an Rnase III-deficient E. coli strain with IPTG-inducible T7 Polymerase activity. To induce dsRNA production from these plasmids, the HT115 bacteria is grown on special RNAi NGM feeding plates that contain IPTG and the ampicillin analog carbenicillin. Carbenicillin is preferred over ampicillin because it tends to be more stable. Accordingly, HT115 strain as a ribonuclease- deficient strains can be utilized to create ribonuclease-deficient and/or ribonuclease-free PLQLFHOOV^^7KH^'(^^GHVLJQDWLRQ^PHDQV^WKDW^UHVSHFWLYH^VWUDLQV^FRQWDLQ^WKH^^'(^^O\VRJHQ^WKDW^ carries the gene for T7 RNA polymerase under control of the lacUV5 promoter. IPTG is required to maximally induce expression of the T7 RNA polymerase in order to express recombinant genes cloned downstream of a T7 promoter. HT115 (DE3) is suitable for expression from a T7 or T7-lac promoter or promoters recognized by the E.coli RNA polymerase: e.g. lac, tac, trc, ParaBAD, PrhaBAD and also the T5 promoter. The genotype of HT115 (DE3) is: F-, mcrA, mcrB, IN(rrnD-rrnE)1, rnc14::Tn10(DE3 lysogen: lavUV5 promoter -T7 polymerase) (IPTG-inducible T7 polymerase) (RNAse III minus). This strain grows on LB or 2XYT plates. This strain is tetracycline resistant. Researchers using this strain can test for expression by transforming in one of the plasmids from the Fire Vector Kit (1999) (pLT76, e.g.) using standard CaCl₂ transformation techniques. This strain is resistant to tetracycline, and can be cultivated at ^^^^ LB, and aerobic. Researchers also use this strain to test the interference experiment of nematodes. [00176] In some embodiments, ribonuclease-deficient minicells disclosed herein are produced from ribonuclease-deficient parental strains including, but are not limited to, HT115 (DE3). In other embodiments, HT115 (DE3) strain is genetically engineered by deleting, 43 303137700 mutating, knocking out, or disrupting minC, minD, and/or minC and minD gene(s) to induce minicell production. In other embodiments, HT115 (DE3) strain is genetically engineered by overexpressing ftsZ and/or minE genes to induce minicell production. [00177] In some embodiments, ribonuclease-deficient minicells disclosed herein can be produced from protease-deficient parental strains including, but are not limited to, BL21 (DE3), BL21-AI and LPS-modified BL21 (DE3), genetically engineered by deleting, mutating, knocking out, or disrupting gene(s) encoding ribonuclease III. In other embodiments, BL21 (DE3), BL21-AI and LPS-modified BL21 (DE3) strains, in which ribonuclease III expression is suppressed, disrupted and/or nullified, are further genetically engineered by deleting, mutating, knocking out, or disrupting minC, minD, and/or minC and minD gene(s) to induce minicell production. In other embodiments, BL21 (DE3), BL21-AI and LPS-modified BL21 (DE3) strains, in which ribonuclease III expression is suppressed, disrupted and/or nullified, are also genetically engineered by overexpressing ftsZ and/or minE genes to induce minicell production. [00178] The present disclosure teaches genotypes of newly-generated ribonuclease-deficient minicell strains comprising i) minC-deleted and ribonuclease III-deleted BL21(DE3); fhuA2 >ORQ@^ RPS7^ JDO^ ^^^ '(^^^ >GFP@^ ¨KVG6 ^^ '(^^ ^ ^^ V%DP+,R^ ¨(FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^ L^^^ ¨QLQ^^ ¨PLQ& rnc14::Tn10, ii) minD-deleted and ribonuclease III-deleted BL21(DE3); IKX$^^>ORQ@^RPS7^JDO^^^^'(^^^>GFP@^¨KVG6 ^^'(^^ ^^^ sBamHIo ¨(FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^ L^^^ ¨QLQ^^ ¨PLQ' rnc14::Tn10, iii) minC/D-deleted and ribonuclease III-deleted BL21(DE3); IKX$^^ >ORQ@^ RPS7^ JDO^ ^^^ '(^^^ >GFP@^¨ KVG6 ^^'(^^ ^^^V%DP+,R^¨ (FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^L^^^¨ QLQ^^¨ PLQ&^ ¨PLQ' rnc14::Tn10, iv) minC-deleted and ribonuclease III-deleted BL21-AI; F- ompT hsdS_B (r_B- m_B-) gal dcm araB::T7RNAP-tetA ¨PLQ& rnc14::Tn10, v) minD-deleted and ribonuclease III-deleted BL21-AI; F-ompT hsdS_B (r_B- m_B-) gal dcm araB::T7RNAP-tetA ¨PLQ' rnc14::Tn10, vi) minC/D-deleted and ribonuclease III-deleted BL21-AI; F- ompT hsdS_B (r_B- m_B-) gal dcm araB::T7RNAP-tetA ¨PLQ&^¨PLQ' rnc14::Tn10; vii) minC- deleted LPS-modified and ribonuclease III-deleted BL21(DE3); PVE$^^^^¨ JXW4^¨ NGV'^¨ OS[/^ ¨OS[0^ ¨SDJ3^ ¨OS[3^ ¨HSW$^ ¨minC rnc14::Tn10, viii) minD-deleted LPS-modified and ribonuclease III-deleted BL21(DE3); PVE$^^^^ ¨JXW4^ ¨NGV'^ ¨OS[/^ ¨OS[0^ ¨SDJ3^ ¨OS[3^ ¨HSW$^¨minD rnc14::Tn10, ix) minC/D-deleted LPS-modified and ribonuclease III-deleted BL21(DE3); PVE$^^^^ ¨JXW4^ ¨NGV'^ ¨OS[/^ ¨OS[0^ ¨SDJ3^ ¨OS[3^ ¨HSW$^ ¨minC, ¨minD rnc14::Tn10, x) minC-deleted and ribonuclease III-deleted B8 with suppression on T7 RNA polymerase activity; IKX$^^>ORQ@^RPS7^JDO^^^^'(^^^>GFP@^¨ KVG6 ^^'(^^ ^^^V%DP+,R^¨ (FR5,- 44 303137700 B int::(lacI::PlacUV5::T7 JHQH^^^ L^^^ ¨QLQ^^ ¨PLQ& rnc14::Tn10; xi) minD-deleted and ribonuclease III-deleted B8 with suppression on T7 RNA polymerase activity; fhuA2 [lon] RPS7^JDO^^^^'(^^^>GFP@^¨ KVG6 ^^'(^^ ^^^V%DP+,R^¨ (FR5,-B int::(lacI::PlacUV5::T7 gene1) L^^^ ¨QLQ^^ ¨PLQ' rnc14::Tn10; xii) minC/D-deleted and ribonuclease III-deleted B8 with suppression on T7 RNA polymerase activity; IKX$^^>ORQ@^RPS7^JDO^^^^'(^^^>GFP@^¨KVG6 ^^ '(^^ ^ ^^ V%DP+,R^ ¨(FR5,-B int::(lacI::PlacUV5::T7 JHQH^^^ L^^^ ¨QLQ^^ ¨PLQ&^ ¨PLQ' rnc14::Tn10; xiii) minC-deleted HT115 (DE3); F-, mcrA, mcrB, IN(rrnD-rrnE)1, rnc14::Tn10(DE3 lysogen: lavUV5 promoter -T7 polymerase) ¨PLQ&^^ [iv) minD-deleted HT115 (DE3); F-, mcrA, mcrB, IN(rrnD-rrnE)1, rnc14::Tn10(DE3 lysogen: lavUV5 promoter -T7 polymerase) ¨PLQ'^^DQG^[Y^ minC/D-deleted HT115 (DE3); F-, mcrA, mcrB, IN(rrnD- rrnE)1, rnc14::Tn10(DE3 lysogen: lavUV5 promoter -T7 polymerase) ¨PLQ&^¨PLQ'^ [00179] Minicells that have segregated from parent cells lack chromosomal and/or nuclear components, but retain the cytoplasm and its contents, including the cellular machinery required for protein expression. In some embodiments, minicells are ribonuclease-deficient because the parent cells are ribonuclease-deficient strains. Although chromosomes do not segregate into minicells, extrachromosomal and/or episomal genetic expression elements may segregate, or may be introduced into minicells after segregation from parent cells. In some embodiments, the disclosure is drawn to ribonuclease-deficient minicells comprising an expression element, which may be an inducible expression element. The inducible expression element such as an inducible promoter can be introduced to a recombinant plasmid used for homologous recombination to knock out and/or delete gene(s) involved to cell division and/or chromosomal partitioning such as minC, minD, and minC/D, a recombinant expression vector to overexpress gene(s) involved to cell division and/or chromosomal partitioning such as ftsZ and minE, and a recombinant expression vector for expressing an enzymatically active polypeptide including a protein of interest disclosed herein. In further embodiments, the inducible expression element comprises expression sequences operably linked to an open reading frame (ORF) that encodes proteins of interest disclosed herein. Optionally, at any point in the method, an inducing agent is provided in order to induce expression of an ORF that encodes proteins of interest disclosed herein. [00180] In some embodiments, the disclosure teaches methods of making a ribonuclease- deficient bacterial minicell comprising a recombinant fusion protein that is not naturally found in parental cells. In some embodiment, the disclosure teaches method of preparing ribonuclease-deficient minicells from the host cells. 45 303137700 [00181] In further embodiments, a minicell is produced from an eukaryotic cell. In further embodiments, the minicell produced as described above is used as an anucleated cell-based platform and/or an agricultural composition for the encapsulation and delivery of biologically active compounds. [00182] In some embodiments, minicells taught in the present disclosure is protease deficient or ribonuclease deficient. In some embodiments, said minicell is protease deficient. In some embodiments, said minicell is ribonuclease deficient. In some embodiments, said minicell is protease deficient and ribonuclease deficient. In some embodiments, said minicell is ribonuclease-deficient, and wherein said biologically active compound is a nucleic acid. In some embodiments, said biologically active compound is said nucleic acid is selected from the group consisting of an antisense nucleic acid, a double-stranded RNA (dsRNA), a short-hairpin RNA (shRNA), a small-interfering RNA (siRNA), a microRNA (miRNA), an antisense RNA, a ribozyme, an aptamer, and combination thereof. Minicell Separation and Purification [00183] A variety of methods are used to separate minicells from parent cells (i.e., the cells from which the minicells are produced) in a mixture of parent cells and minicells. In general, such methods are physical, biochemical and genetic, and can be used in combination. (i) Physical Separation of Minicells from Parent Cells [00184] By way of non-limiting example, minicells are separated from parent cells glass-fiber filtration (Christen et al., Gene 23:195-198, 1983), and differential and zonal centrifugation (Barker et al., J. Gen. Microbiol.111:387-396, 1979), size-exclusion chromatography, e.g. gel- filtration, differential sonication (Reeve, J. N., and N. H. Mendelson.1973. Biochem. Biophys. Res. Commun. 53:1325-1330), and UV-irradiation (Tankersley, W. G., and J. M. Woodward. 1973. Proc Soc Exp Biol Med. 1974 Mar;145(3):802–805). [00185] Some techniques involve different centrifugation techniques, e.g., differential and zonal centrifugation. By way of non-limiting example, minicells may be purified by the double sucrose gradient purification technique described by Frazer and Curtiss, Curr. Topics Microbiol. Immunol.69:1-84, 1975. [00186] Other physical methods may also be used to remove parent cells from minicell preparations. By way of non-limiting example, mixtures of parent cells and minicells are frozen WR^í^^^^&^^DQG^WKHQ^WKDZHG^VORZO\^^Frazer and Curtiss, Curr. Topics Microbiol. Immunol.69:1- 84, 1975). 46 303137700 (ii) Biochemical Separation of Minicells from Parent Cells [00187] Contaminating parental cells may be eliminated from minicell preparations by incubation in the presence of an agent, or under a set of conditions, that selectively kills dividing cells. Because minicells can neither grow nor divide, they are resistant to such treatments. [00188] Examples of biochemical conditions that prevent or kill dividing parental cells is treatment with an antibacterial agent, such as penicillin or derivatives of penicillin. Penicillin has two potential affects. First, penicillin prevent cell wall formation and leads to lysis of dividing cells. Second, prior to lysis dividing cells form filaments that may assist in the physical separation steps described above. In addition to penicillin and its derivatives, other agents may be used to prevent division of parental cells. Such agents may include azide. Azide is a reversible inhibitor of electron transport, and thus prevents cell division. As another example, D-cycloserine or phage MS2 lysis protein may also serve as a biochemical approach to eliminate or inhibit dividing parental cells. (Markiewicz et al., FEMS Microbiol. Lett.70:119- 123, 1992). Khachatourians (U.S. Pat. No.4,311,797) states that it may be desirable to incubate PLQLFHOO^SDUHQW^ FHOO^PL[WXUHV^ LQ^ EUDLQ^ KHDUW^ LQIXVLRQ^ EURWK^ DW^ ^^^^&^^ WR^ ^^^^&^^ SULRU^ WR^ WKH^ addition of penicillin G and further incubations. (iii) Genetic Separation of Minicells from Parent Cells [00189] Alternatively or additionally, various techniques may be used to selectively kill, preferably lyse, parent cells. For example, although minicells can internally retain M13 phage in the plasmid stage of the M13 life cycle, they are refractory to infection and lysis by M13 phage (Staudenbauer et al., Mol. Gen. Genet.138:203-212, 1975). In contrast, parent cells are infected and lysed by M13 and are thus selectively removed from a mixture comprising parent cells and minicells. A mixture comprising parent cells and minicells is treated with M13 phage at an M.O.I.=5 (phage cells). The infection is allowed to continue to a point where ؤ50% of the parent cells are lysed, preferably ؤ75%, more preferably ؤ95% most preferably ؤ99%; and أ25% of the minicells are lysed or killed, preferably أ15%, most preferably أ1%. [00190] As another non-limiting example of a method by which parent cells can be selectively killed, and preferably lysed, a chromosome of a parent cell may include a conditionally lethal gene. The induction of the chromosomal lethal gene will result in the destruction of parent cells, but will not affect minicells as they lack the chromosome harboring the conditionally lethal gene. As one example, a parent cell may contain a chromosomal integrated bacteriophage comprising a conditionally lethal gene. One example of such a bacteriophage is an integrated 47 303137700 lambda phage that has a temperature sensitive repressor gene (e.g., lambda cI857). Induction of this phage, which results in the destruction of the parent cells but not of the achromosomal minicells, is achieved by simply raising the temperature of the growth media. A preferred bacteriophage to be used in this method is one that kills and/or lyses the parent cells but does not produce infective particles. One non-limiting example of this type of phage is one that lyses a cell but which has been engineered to as to not produce capsid proteins that are surround and protect phage DNA in infective particles. That is, capsid proteins are required for the production of infective particles. [00191] As another non-limiting example of a method by which parent cells can be selectively killed or lysed, toxic proteins may be expressed that lead to parental cell lysis. By way of non- limiting example, these inducible constructs may employ a system to control the expression of a phage holing gene. Holin genes fall with in at least 35 different families with no detectable orthologous relationships (Grundling, A., et al. 2001. Proc. Natl. Acad. Sci. 98:9348-9352) of which each and any may be used to lyse parental cells to improve the purity of minicell preparations. [00192] In some embodiments, minicells are substantially separated from the minicell- producing parent cells in a composition comprising minicells. After separation, the compositions comprising the minicells is at least about 99.9%, about 99.8%, about 99.7%, about 99.6%, about 99.5%, about 99.4%, about 99.3%, about 99.2%, about 99.1%, about 99%, about 98%, about 97%, about 96%, about 95%, about 94%, about 93%, about 92%, about 91%, about 90%, about 89%, about 88%, about 87%, about 86%, about 85%, about 84%, about 83%, about 82%, about 81%, about 80%, about 79%, about 78%, about 77%, about 76%, about 75%, about 74%, about 73%, about 72%, about 71%, about 70%, about 65%, about 60%, about 55%, about 50%, about 45%, about 40%, about 35%, about 30%, about 25% or about 20% free of minicell-producing parent cells. Thus, the compositions of the disclosure can comprise minicells that are substantially free of the parental cell. [00193] In some aspects, the present invention provides a method for making minicells, the method comprising (a) culturing a minicell-producing parent cell, wherein the parent cell comprises an recombinant construct, wherein the recombinant construct comprises a nucleotide sequence homologous to a target gene associated with regulating cell division, and (b) separating the minicells from the parent cell, thereby generating a composition comprising minicells. In some embodiments, the method further comprises (c) purifying the minicells from the composition by centrifugation and/or filtration. In some embodiments, one or more additional expression constructs can be introduced into the minicell-producing parent cell 48 303137700 which comprise genes associated with cell division. In such instances, the expression constructs may be simultaneously or sequentially introduced into the parent cell prior to induction for minicell formation, and can comprise one or more selection markers (e.g., antibiotic resistance genes) and/or reporter genes to allow for selection and/or visualization of minicells expressing the protein(s) of interest. In other embodiments, the expression construct can express one or more additional proteins, which are driven by the same or different promoters, including inducible promoters. In further embodiments, genes associated with cell division are minC, minD, and/or both minC and minD. Encapsulation [00194] Encapsulation is a process of enclosing the substances within an inert material, which protects from environment as well as control release of active compounds. Two type of encapsulation has been well studies; 1) Nanoencapsulation that is the coating of various substances within another material at sizes on the nano scale, and 2) Microencapsulation that is similar to nanoencapsulation aside from it involving larger particles and having been done for a greater period of time than nanoencapsulation. Encapsulation is a new technology that has wide applications in pharmaceutical industries, agrochemical, food industries and cosmetics. In some embodiments, at least one biologically active compound described herein is inert to a cell other than a cell of a target. [00195] In some embodiments, a minicell is utilized to encapsulate and deliver biologically active compounds. The bacterial cells including gram-negative bacteria, gram-negative bacteria, and Extremophilic bacteria, can produce the platform, which can encapsulate the desired biologically active compounds. The anucleated cells comprises minicells that are produced from parental bacterial cells disclosed herein naturally and/or by genetic engineering techniques taught herein. [00196] The present disclosure teaches the benefit of using bacterial minicells which simplify purification of minicell and reduce costs of encapsulation thereof. By employing encapsulation to biologically active compounds, the compounds are protected from external factors that causes degradation of the compounds and reduces life cycle of the compounds. [00197] Current encapsulation techniques include oils, invert suspensions, polymer-based nanomaterials, lipid-based nanomaterials, porous inorganic nanomaterials, and clay-based nanomaterials. [00198] COC (Crop Oil Concentrate) and MSO (Methylated Seed Oil) technologies are used for oil encapsulation. They act as humectants to move the active ingredient droplets through 49 303137700 the spray nozzle and reconfigure the droplets on the outside to keep the active ingredients from evaporating. [00199] Invert suspension is an oil sub-category providing either a suspension of water encapsulated within an oil shell or water surrounded by an oil coating used to minimize the creation of driftable fines (sub 105 microns) after being sprayed through a nozzle tip. This technology works on reducing driftable fines for the active ingredients. [00200] Polymer-based nanomaterials consist of a polymer that has nanoparticles or nanofillers dispersed within the polymer matrix. Typically, the polymers are contrasting (one hydrophobic, one hydrophilic) to sustain amphiphilic properties. Either synthetic or natural polymers (guar gum) act to increase the viscosity of the spray solution and affect the rheological profile by producing larger spray particles. Polymer-based adjuvants increase the possibility of spray particles shattering, increasing drift. However, use of polymer-based drift reduction technology adjuvants for aerial applications of active ingredients is not recommend. Although they have an efficient loading capacity, the necessary polymers are expensive, limiting scalability. [00201] Lipid-based nanomaterials have great potential to encapsulate hydrophilic, hydrophobic, and lipophilic active ingredients, and are commonly used in the pharmaceutical field. However, scalable production is significantly limited by cost. [00202] Porous inorganic nanomaterials, such as silica nanoparticles, are effective at encapsulating bioactive molecules, but face limitations in biodegradability and scalability. These polymer-coated nanoparticles suffer from various limitations such as poor thermal and chemical stability, rapid elimination by the plant enzyme system, and degradation of some polymers, resulting in the formation of acidic monomers and decreased pH value within the polymer matrix. Clay nanoparticles are economically viable and provide great opportunities for developing multifunctional nanocarrier materials, but are energy intensive, requiring high heat for production. These alternatives cannot be modified as easily to provide targeted delivery to plants. [00203] In some embodiments, a minicell and/or an agricultural composition comprising the minicell has advantages in cost and biodegradability. The minicell platforms are easily scaled through common, industrial fermentation practices. Once scaled, they can be purified through a series of centrifugation and/or filtration steps. The self-assembly of the carbohydrate-binding modules to the surface of minicells significantly cuts the cost of making a targeting bioparticle. Additionally, an anucleated cell-based minicell platform is advantageous compared to other encapsulation technologies in terms of biocompatibility for plant and environmental use; this 50 303137700 is because the anucleated cell-based minicell platform is derived by safe, commonly found microbes that are native to the applied areas and can safely biodegrade to be reused by the ecosystem. This platform suitable for scalable, non-toxic delivery can play an significant role in the field of agriculture. [00204] In order to solve problems of conventional agrochemicals or biologicals that are easily degraded or evaporated before they reach their intended target, the present disclosure provides a minicell and/or an agricultural composition for the encapsulation and delivery of biologically active compounds aims to protect the bioactivity from external factors until the compounds are applied to a target and to be slowly released to the intended target. The various mechanisms by which biologically active compounds are typically lost to the environment are averted using the disclosed minicell-based encapsulation and delivery platform. This is because the lipid-bilayer of the minicell acts as an effective layer of protection against harsh environmental conditions. Specifically, the internalization of the active inside of the minicell protects the compounds against sharp changes in temperature, pH, or strong exposure to light. In other words, the minicell protects the compounds against volatilization, photolytic degradation, and hydrolysis. Therefore, the biologically active compounds can remain protected from adverse external factors and is allowed for gradual and/or controlled release to intended targets via minicell-based platform that encapsulates the biologically active compound of interest. [00205] Furthermore, the other benefit of the present disclosure provides a minicell and/or an agricultural composition for the encapsulation and delivery of biologically active compounds is that this platform offers the improved and enhanced targeting capability to the plant and its microenvironment. The inherent surface chemistry of the outer membrane of the minicell- based bioparticle naturally mimics that of bacteria. This is significant because there are many types of bacteria that live symbiotically in a microbiome on the surface of plant leaves, stems, and in their root system. By using the minicell-based platform, biological membrane of the minicell has natural adherence to the various surfaces of plants. This feature allows for delivering encapsulated biologically-active compounds including biocontrols and biostimulants in the minicell chassis that is targeted to adhere to plant surfaces and the soil microenvironment around the plant’s root system as well as to other targets such as pests, insects, bugs, weeds, worms, bacteria, viruses, pathogens, and parasites. In addition to relying on the natural adherence of the minicell-based bioparticle to plants, the present disclosure teaches uses of genetic engineering to give rise to surface-expressing moiety fused with specific 51 303137700 binding domain on the membrane of the minicell. In this way its ability to target the plant or the pest is significantly enhanced. [00206] In some embodiments, the present disclosure provides the genetic engineering techniques to make minicell-based platform with binding domains/motifs that functionalize the surface of the minicell. Proteins including specific binding domains and/or motifs are expressed on the surface of the minicells and specifically target binding sites that are present on the surface of plants or pests. [00207] In some embodiments, minicell-based platform can be functionalized by proteins with carbohydrate binding modules (CBMs) that can target and bind to carbohydrates such as cellulose, xylan, chitin, and lignin, which are important and ubiquitous structural components of plant cell walls. Because CBMs can recognize their binding site present on a subject such as a plant or a pest, the minicell-based platform comprising the functionalized binding domain allows for targeting with high specificity. [00208] In some embodiments, the use of CBMs is not limited to agriculture uses. CBMs can be used for the purification of active ingredients or biomolecules through the means of cellulose columns. Supplementary to the surface chemistry of the minicell-based platform, the relative mass of the bioparticle can also significantly mitigate the off-target exposure of active compounds due to aerosolization and leaching. By concentrating and encapsulating actives in the relatively large chassis of the minicell before being sprayed, the compound is less susceptible to aerosolization or drift caused by wind when compared to spraying free-floating compounds. Furthermore, the larger size of the minicell encapsulation and delivery platform can mitigate the leaching of actives through the soil and into groundwater supplies. Agriculturally Acceptable Carrier [00209] Compositions described herein can comprise an agriculturally acceptable carrier. The composition useful for these embodiments may include at least one member selected from the group consisting of a tackifier, a microbial stabilizer, a fungicide, an antibacterial agent, a preservative, a stabilizer, a surfactant, an anti-complex agent, an herbicide, a nematicide, an insecticide, a plant growth regulator, a fertilizer, a rodenticide, a desiccant, a bactericide, a nutrient, or any combination thereof. In some examples, compositions may be shelf-stable. For example, any of the compositions described herein can include an agriculturally acceptable carrier (e.g., one or more of a fertilizer such as a non-naturally occurring fertilizer, an adhesion agent such as a non- naturally occurring adhesion agent, and a pesticide such as a non-naturally occurring pesticide). A non-naturally occurring adhesion agent can be, for example, a polymer, 52 303137700 copolymer, or synthetic wax. For example, any of the coated seeds, seedlings, or plants described herein can contain such an agriculturally acceptable carrier in the seed coating. In any of the compositions or methods described herein, an agriculturally acceptable carrier can be or can include a non-naturally occurring compound (e.g., a non-naturally occurring fertilizer, a non-naturally occurring adhesion agent such as a polymer, copolymer, or synthetic wax, or a non-naturally occurring pesticide). [00210] In some embodiments, a minicell described herein can be mixed with an agriculturally acceptable carrier. The carrier can be a solid carrier or liquid carrier, and in various forms including microspheres, powders, emulsions and the like. The carrier may be any one or more of a number of carriers that confer a variety of properties, such as increased stability, wettability, or dispersability. Wetting agents such as natural or synthetic surfactants, which can be nonionic or ionic surfactants, or a combination thereof can be included in the composition. Water-in-oil emulsions can also be used to formulate a composition that includes the isolated bacteria (see, for example, U.S. Patent No. 7,485,451). Suitable formulations that may be prepared include wettable powders, granules, gels, agar strips or pellets, thickeners, liquids such as aqueous flowables, aqueous suspensions, water-in-oil emulsions, etc. The formulation may include grain or legume products, for example, ground grain or beans, broth or flour derived from grain or beans, starch, sugar, or oil. [00211] In some embodiments, the agricultural carrier may be soil or a plant growth medium. Other agricultural carriers that may be used include water, fertilizers, plant-based oils, humectants, or combinations thereof. Alternatively, the agricultural carrier may be a solid, such as diatomaceous earth, loam, silica, alginate, clay, bentonite, vermiculite, seed cases, other plant and animal products, or combinations, including granules, pellets, or suspensions. Mixtures of any of the aforementioned ingredients are also contemplated as carriers, such as but not limited to, pesta (flour and kaolin clay), agar or flour-based pellets in loam, sand, or clay, etc. Formulations may include food sources for the bacteria, such as barley, rice, or other biological materials such as seed, plant parts, sugar cane bagasse, hulls or stalks from grain processing, ground plant material or wood from building site refuse, sawdust or small fibers from recycling of paper, fabric, or wood. [00212] Additional examples of agriculturally acceptable carriers include dispersants (e.g., polyvinylpyrrolidone/vinyl acetate PVPIVA S-630), surfactants, binders, and filler agents. [00213] Persons having skill in the art will appreciate that, unless otherwise noted, all references to an agricultural composition in the present disclosure can be read as referring to an agricultural formulation. Therefore, embodiments described in the present disclosure which 53 303137700 refer to an agricultural composition will also be understood to refer to an agricultural formulation. Release of Biologically Active Compounds Encapsulated by Minicell [00214] The present disclosure teaches that substances (including nucleic acids, RNA molecules, or agrochemicals) is retained within the minicell and be released over time. The disclosure teaches a high value, low volume product of a minicell encapsulating at least one biologically active compounds and/or expressing a fusion protein. In some embodiments, the fusion protein has at least one surface expressing moiety and at least one cell adhesion moiety. In some embodiments, the fusion protein has at least one surface expressing moiety and at least one cell stimulation moiety. In some embodiments, the fusion protein has at least one surface expressing moiety and at least one cell degrading moiety. In some embodiments, the anucleated cell-based product can be sprayed much less than other commercially available agrochemical products and also retain the desired effects of the active compounds over a longer period of time. [00215] The term “controlled release” as used herein means that one or more substances (including nucleic acids, RNA molecules, or agrochemicals) encapsulated by a minicell described in the present disclosed is released over time in a controlled manner. The controlled release is meant for purposes of the present disclosure that, once the biologically active compound is released from the formulation, it is released at a controlled rate such that levels and/or concentrations of the compounds are sustained and/or delayed over an extended period of time from the start of compound release, e.g., providing a release over a time period with a prolonged interval. [00216] Current controlled release mechanism is based mainly on fully encapsulation of fertilizer (e.g. Agrium, ICL, Kingenta and Ekompany) or pesticides (e.g. Adama, Syngenta, Bayer). Fully encapsulation of fertilizer is usually based on resins (e.g. polyurethanes) or sulfur base mixture. Pesticides are loaded into micro polymeric capsules. Products of encapsulated fertilizer are limited to milligrams scale of dry fertilizer, due to the need of thick wall opposing the high inner pressure. This pressure is built up due to water entering the capsule driven by the negative osmotic potential of the dissolve fertilizer. As more fertilizer is encapsulated, more pressure will build up and a thicker wall is required. The feasible ratio between fertilizer amounts to wall thickness is in the tens of milligrams scale. Nevertheless encapsulated fertilizer is still very expensive and costs up to four times over the fertilizer price. 54 303137700 [00217] Moreover, the release mechanism is based on transport through faults and cracks distributed in the casing. Meaning, coating must be uniform throughout the all surface area, which is in turn a manufacturing challenge. On top of that, the materials being used for coating are temperature sensitive and change their structural properties extremely in small temperature UDQJH^^^^^^&^-^^^^&^^^^OHDGLQJ^WR^UDGLFDO^FKDQJHV^LQ release rates (up to double the rate). Thus, conventional encapsulation of agrochemicals and biologicals has challenges of uniform coating and temperature dependent. [00218] If it is desired to permit fast release of the encapsulated composition during drying of the formulation on a leaf, or similar, surface it is necessary to have thin walled microcapsules. Typically microcapsules with a mean diameter of about 2 microns require a polymer wall concentration in the formulation of about 3 % by weight. Greater quantities of polymer will slow the release rate. The diameter of the capsules and the quantity of wall forming polymer can be used to tune the performance of the capsules, depending on the required pesticide and the conditions of use. [00219] The increasing use of pesticides, herbicides, fungicides, insecticides, nematicides, fertilizer and the like, poses serious health and environmental problems which must be controlled in order to minimize the harmful effects of those products. One problem frequently encountered with herbicides, such as alachlor, metolachlor, norflurazon and sulfometuron is leaching and migration, which results in loss of herbicidal efficiency and can cause damage to other crops and contaminate water. [00220] The present disclosure teaches that biologically active compounds encapsulated by minicells disclosed herein can be released in a controlled manner. In some embodiments, the controlled release of the compounds are determined by a treatment of an agent such as glutaraldehyde, formaldehyde, as well as natural compounds, such as genipin, and epigallocatechin gallat, derivatives of ethylene glycol di(meth)acrylate, derivatives of methylenebisacrylamide, and formaldehyde-free crosslinking agent DVB (Divinyl Benzene). In some embodiments, a varying concentration of the agent (e.g. glutaraldehyde) can prevent the degradation of minicells encapsulating the biologically active compounds in different degrees. [00221] In other embodiments, the agent includes, but is not limited to glutaraldehyde, formaldehyde, as well as natural compounds, such as genipin, and epigallocatechin gallat, derivatives of ethylene glycol di(meth)acrylate, derivatives of methylenebisacrylamide, and formaldehyde-free crosslinking agent DVB (Divinyl Benzene). 55 303137700 [00222] In some embodiments, biologically active compounds encapsulated by minicells disclosed herein can be released at a rate of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of a desired minicell unit/input per day. In other embodiments, an amount of the desired minicell unit/input accounts for encapsulated biologically active compounds. Encapsulation amount of biologically active compounds can calculate encapsulation fraction and mass fraction, which determines the desired minicell unit and/or input per day. [00223] In some embodiments, minicells without treatment of an agent (e.g. glutaraldehyde) may have an initial fast release of 10%, 20%, 30%, 40%, 50%, 60%, 70%, or 80% of their desired unit/input per day and are followed by a controlled release of minicells treated with a varying concentration of the agent (e.g. glutaraldehyde), which give rise to a controlled release of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% of the desired input per day. In some embodiments, a varying concentration of the agent (e.g. glutaraldehyde) can prevent the degradation of minicells encapsulating the biologically active compounds in different degrees. In some embodiments, the agent includes, but is not limited to glutaraldehyde, formaldehyde, as well as natural compounds, such as genipin, and epigallocatechin gallat, derivatives of ethylene glycol di(meth)acrylate, derivatives of methylenebisacrylamide, and formaldehyde-free crosslinking agent DVB (Divinyl Benzene). Improved Encapsulation and Retention [00224] In order to improve encapsulation and retention, the present disclosure teaches that solvents can be used in the encapsulation solution to increase the solubility of the biologically active compounds in the minicells. These solvents include, but are not limited to, CaCl₂ solution, ethanol, DMSO, polyethylene glycol, and glycerol. Not only can these solvents be used to increase the solubility of certain active compounds, but they may be used to increase the diffusion of the active compounds into the cell through certain protein channels or through the lipid bilayer of the outer membrane. In addition to the use of solvents to enhance the encapsulation process of the minicell, certain fixatives, preservatives, and cross-linking agents can be used to trap the active ingredient within the membrane of the minicell, cross-link certain active compounds to the minicell itself, and improve the stability of the minicell. The relative concentration of these stabilizing/cross-linking agents can be tuned to achieve the required 56 303137700 loading capacity for the active ingredient as well as the release kinetics of the active ingredient from the cell. These agents include, but are not limited to synthetic compounds, such as glutaraldehyde, formaldehyde, as well as natural compounds, such as genipin, and epigallocatechin gallat. [00225] In some embodiments, minicells described herein are treated with a solvent, agent, fixative, preservative, or cross-linking agent for better solubility, increased stability, or enhanced integrity. In some embodiments, said minicell exhibits a controlled release rate of said biologically active compound, wherein the release can be a steady release or an initial burst followed by steady release. [00226] In other embodiments, minicells can show their innate and modified stability and can withstand various environmental conditions and changes in temperature, pH, and/or shear stress. [00227] In further embodiments, the present disclosure teaches that the minicell can be derived from ribonuclease-deficient cell strains and/or protease-deficient cell strains. Also, the minicell can be generated from cell strains genetically engineered to disrupt structure/function of ribonuclease and/or protease. The ribonuclease-deficient minicell can capture and deliver dsRNAs to a target disclosed herein. In order to enhance encapsulation and retention of dsRNAs, the present disclosure teaches expression of dsRNA binding protein internally and/or externally. Once the dsRNA binding protein recognizes and binds to the dsRNA within the minicells, the dsRNA cannot flow back across the membrane. Also, the dsRNA binding protein can aid in dsRNA encapsulation and retention as well as protect dsRNA from degradation by RNase. On the other hand, the protease-deficient minicell can better encapsulate and retain dsRNA within the minicells when the dsRNA binding protein is expressed to protect dsRNA from RNase activity. RNase cannot have an easy access to the dsRNA bound to the dsRNA binding protein for degradation. The dsRNA binding protein can also be expressed in conjunction with internal dsRNA production to ensure better retention. [00228] In some embodiments, the minicell expresses a polypeptide within the cell, and wherein the polypeptide binds to said at least one biologically active compound such as dsRNA within the cell. In other embodiments, said at least one biologically active compound is a dsRNA and wherein said polypeptide is a dsRNA binding protein. The dsRNA binding protein increases stability of said dsRNA and protects said dsRNA from degradation. In further embodiments, the dsRNA binding protein is DRB4 protein. In some embodiments, an agricultural formulation comprises a polypeptide within minicells, wherein said polypeptide is expressed within said minicell, wherein said polypeptide binds to said nucleic acid. In some 57 303137700 embodiments, said polypeptide is a dsRNA binding protein, and wherein said dsRNA binding protein increases loading and enhances the stability of dsRNA. Invasive Delivery [00229] The present disclosure teaches an invasive delivery method of biologically active compounds into a target cell, which is not a mammalian cell by application of an agent that can help improve penetration of the minicell into targets such as plants, pests, insects, bugs, worms, pathogens and parasites. The minicells encapsulating the biologically active compounds described herein is applied to a target cell with an agent. In some embodiments, the agent is an adjuvant for improving penetration of the minicell into the target cell and invasively delivering the biologically active compounds within the target cell. The agent is a surfactant, an emulsifier, a crop oil concentrate, a penetrant, a salt or combination thereof. Not-limiting examples of the agent are methylated seed oil, N,N-dimethyldecanamide, and N-decyl-N- methly formamide. In some embodiments, a method of delivering at least one biologically active compound is provided, comprising: applying said minicell to said target cell with an agent, wherein said agent is an adjuvant for improving penetration of minicells into a target cell. In further embodiments, a method of delivering at least one biologically active compound is provided, said agent is a surfactant, an emulsifier, a crop oil concentrate, a penetrant, a salt or combination thereof . [00230] Various surfactants and other formulation additives can be used to enhance the uptake/invasiveness of nanoparticles or compounds into plants through the roots and leaves. Silicone surfactants can enhance the uptake of compounds and nanoparticles through the stomata, cuticle, and root system. Lipid-based liquid crystalline nanoparticles can be used as a surfactant to improve delivery of biologically active compounds through the cuticle layer. [00231] In other embodiments, the present disclosure teaches an invasive delivery method of biologically active compounds into a target cell by expressing proteins that improve penetration of plant surface or increase uptake through the roots or stomata. In some embodiments, the minicells express at least one fusion protein comprises at least one surface expressing moiety and at least one target cell degradation moiety. The target cell degradation moiety comprises an cutinase and cellulose, which can facilitate minicells to pass through plant surface and deliver biologically active compounds into a target cell, tissue or organ. [00232] In some embodiments, the intact minicell has a cutinase on its surface that facilitate said minicell to penetrate through a plant cuticle into the target cell. The intact minicell expresses a heterologous cutinase that is displayed on its surface. The intact minicell has a 58 303137700 cellulase on its surface that breaks down a target cell wall and facilitate said minicell to penetrate into the target cell. The intact minicell has a heterologous cellulase that is displayed on its surface. [00233] In further embodiments, the present disclosure teaches an invasive delivery method of biologically active compounds into a target cell, which is not a mammalian cell, by generating minicells from plant invasive species such as Agrobacterium and Endophytes. [00234] The present disclosure provides compositions and methods of producing minicells from plant pathogenic bacteria and fungi such as endophytes. The bacterial and/or yeast species has mechanisms to transport itself from the environment to the cells, internal tissues or organs of target plants. In some embodiments, minicells from these bacterial and yeast endophytes are produced. The endophytes used for minicell production include, but are not limited to Acidovorax facilis, Bradyrhizobium, Rhizobium, Rhodococcus rhodochrous, Colletotrichum, Curvularia, Epichloë, Fusarium, Mycosphaerella, Neotyphodium, Piriformospora, Serendipita. The minicells derived from endophytes can encapsulate biologically active compounds described herein and deliver them into the internal parts of target plants by invasion/penetration mechanisms. [00235] There are several pathways by which biologically active compounds or particles are able to be uptaken through the leaf. These pathways include through trichomes, stomata, plant wounds, root junctions, stigma, and the cuticle (Alshaal et al., Env. Biodiv. Soil Security 1:71- 83, 2017). Due to the extensive presence of the cuticle at the outermost layer of plant leaves, a primary manner in which foliar uptake occurs is through the cuticle layer. Various compounds, both lipophilic and hydrophilic, are able to transport across the cuticle through aqueous pores (for polar compounds) or cutin matrices (for apolar compounds) (Wang et al., Pesticide Biochemistry and Physiology, 87(1):1-8, 2007). It has been reported that all kinds of nanoparticles, from negatively charged silica nanoparticles (20 nm) to lipid-based liquid crystalline NPs (150-300 nm), have been shown to accumulate above actinal cell walls and in the cuticle (Schwab et al., J of Nanotoxicology 10(3):257-278, 2016). There are permeable regions of the cuticle, such as trichomes, hydathodes, or cell junctions, in plant tissue that have also have uptake functions. [00236] On the other hand, plants are able to uptake compounds and nanoparticles through the stomata. The ability for uptake through the stomata varies for each plant species, but the stomata has generally shown to have a high transport velocity into the leaf, especially for particles or compounds less than 10 nm. However, it is also the case that larger nanoparticles have been able to enter the plant through stomata openings. Foliar application of nanoparticles 59 303137700 has been shown to lead to translocation of nanoparticle from stomatal cavities to plant tissues, the vasculature, and roots cuticle (Schwab et al., J of Nanotoxicology 10(3):257-278, 2016). Bacteria (which are larger than minicells) are also able to invade plants through stomata openings, often times regulating their openings using virulence factors (Zeng et al., Curr. Opin. Biotechnol. 21(5):599-603, 2010). In some embodiments, minicells disclosed herein can be uptaken to target plants and translocated to target cells when the minicells encapsulating biologically active compounds are applied to leaves of target plants. [00237] Agricultural applications of nanoparticles in soil can be very effective since nanoparticles generally accumulate in the first few meters or centimeters of the soil and therefore, interact closely with the rhizosphere. Many studies have shown that nanoparticles are able to accumulate and aggregate near the roots, root tips, root caps, and mucilage of plants. It has also been shown that the mucilage, exudates, and exDNA of plants around its root system serves as a “trap” that immobilizes some nanoparticles and bacteria. Furthermore, plant roots have been shown to be able to uptake and absorb a variety of compounds and nanoparticles into the plant vasculature and tissue (Schwab et al., J of Nanotoxicology 10(3):257-278, 2016). In some embodiments, minicells disclosed herein can be uptaken to target plants and translocated to target cells when the minicells encapsulating biologically active compounds are applied to soil and/or roots of target plants. [00238] Once these compounds and/or nanomaterials have successfully invaded the plant and are in proximity to the plant cell membranes, they can undergo a process of endocytosis. The plant cell membrane uptakes extracellular material, including nanoparticles, through endocytosis. Nanoparticles, up to 500 nm and regardless of charge, can enter the plant cell through endocytosis. Alternative pathways for nanoparticles and other compounds into plant cells are through the permeable pathways of the cell membrane themselves. One of these pathways, aquaporins, allows for non-ionic, solutes to be non-selectively be uptaken into plant cells. In some embodiments, at least one biologically active compound is delivered into a target cell, which is not a mammalian cell, when the minicell described herein is applied by endocytosis. In some embodiments, minicells descried herein are applied to a target and delivered into a cell of a target by endocytosis. Target [00239] As used herein, the term “target” is intended to include any target surface to which a compound, a minicell, an agricultural composition or a minicell of the present disclosure may be applied to a plant or a pest. For example to a plant, plant material including roots, bulbs, 60 303137700 tubers, corms, leaves, flowers, seeds, stems, callus tissue, nuts, grains, fruit, cuttings, root stock, scions, harvested crops including roots, bulbs, tubers, corms, leaves, flowers, seeds, stems, callus tissue, nuts, grains, fruit, cuttings, root stock, scions, or any surface that may contact harvested crops including harvesting equipment, packaging equipment and packaging material. [00240] The term “target cell” refers to cells that is a component of each target. [00241] In some embodiments, exemplary crops, according to certain embodiments of the present disclosures, include but not limited to Row crops, specialty crops, commodity crops, and ornamental crops. Examples of row crops include sunflower, potato, canola, dry bean, field pea, flax, safflower, buckwheat, cotton, maize, soybeans, and sugar beets. Examples of commodity crops include maize, soybean and cotton. Examples of ornamental crops include boxwood, christmas trees, greenhouse grown decorative plants [00242] The present disclosure also teaches exemplary crops as a target, according to certain embodiments of the present disclosure, including vegetables such as broccoli, cauliflower, globe artichoke, peas, beans, kale, collard greens, spinach, arugula, beet greens, bok choy, chard, choi sum, turnip greens, endive, lettuce, mustard, greens, watercress, garlic chives, gai lan, leeks, Brussels sprouts, capers, kohlrabi, celery, rhubarb, cardoon, Chinese celery, lemon gass, asparagus, bamboo shoots, galangal, ginger, soybean, mung beans, urad, carrots parsnips, beets, radishes, rutabagas, turnips, burdocks, onions, shallots, leeks, garlic, green beans, lentils, and snow peas; fruits, such as tomatoes, cucumbers, squash, zucchinis, pumpkins, melons, peppers, eggplant, tomatillos, christophene, okra, breadfruit, avocado, blackcurrant, redcurrant, gooseberry, guava, lucuma, chili pepper, pomegranate, kiwifruit, grapes, cranberry, blueberry, orange, lemon, lime, grapefruit, blackberry, raspberry, boysenberry, pineapple, fig, mulberry, hedge apple, apple, rose hip, and strawberry; nuts such as almonds, pecans, walnuts, brazil nuts, candlenuts, cashew nuts, gevuina nuts, horse-chestnuts, macadamia nuts, Malabar chestnuts, mongongo, peanuts, pine nuts, and pistachios; tubers such as potatoes, sweet potatoes, cassava, yams, and dahlias; cereals or grains such as maize, rice, wheat, barley, sorghum, millet, oats, rye, triticale, fonio, buckwheat, and quinoa; fibers, including, for example, cotton, flax, hemp, kapok, jute, ramie, sisal, and other fibers from plants; stimulant crops, including, for example, coffee, cocoa bean, tea, mate, other plants; and pulses, including, for example, beans (including, for example, kidney, haricot, lima, butter, adzuki, mungo, golden, green gram, black gram, urd, scarlet runner, rice, moth, tepary, lablab, hyacinth, jack, winged, guar, velvet, yam, and other beans), horse-bean, broad bean, field bean, garden pea, chickpea, bengal gram, garbanzo, cowpea, blackeyed pea, pigeon 61 303137700 pea, cajan pea, congo bean, lentil, bambara ground nut, earth pea, vetches, lupins, and other pulses. [00243] In some embodiments, the present disclosure also teaches exemplary aquaculture targets including fish, shrimp, shellfish, and crustacean. The target can be viruses that cause diseases. [00244] The present disclosure teaches that a target cell comprises a plant cell, an insect cell, a worm cell, a bacterial cell, a fungal cell, a virus and a cell of an aquatic animal, wherein said aquatic animal comprises a fish, a shellfish, and a crustacean. [00245] It is appreciated that the minicell and/or agricultural formulation as described herein is particularly useful within the fishing and aquaculture industries, primarily by causing a reduction in the harmful effects of microbial organisms exerted on shellfish, cartilaginous fish, fin fish or aquatic mammals. Shellfish may comprise the group of filter-feeding bivalves such as e.g. clams, oysters, scallops and mussels, and may in addition comprise lobsters, crabs and shrimps. Finfish include, but are not limited to the salmonid species including Atlantic salmon (Salmo salar), rainbow trout (Oncorhynchus mykiss). Further aquatic animal is a fish including a gadid species including Gadus callarias, sea trout (Salmo trutta) and sea bass (Dicentrarchus labrax) and cod, eel as well as fresh water finfish and carp. Further, an aquatic animal may be a dolphin or a whale. [00246] Aquatic animals further encompass any of the broadly known ornamental fish widely used throughout the hobby of fish tank maintenance. Ornamental hobby fish include both fresh water and salt water fish. Representative species of ornamental fish are well known to enthusiasts of the hobby. Preferably the aquatic animal is an animal farmed in an aquaculture. The aquatic animal may be in an early developmental stage e.g., such as larvae and juvenile animals, or a later developmental stage subsequent to the juvenile stage. [00247] The present disclosure provides that the anucleated cell-based platform and/or agricultural formulation as described herein, is targeted to a plant, an insect, a worm, a bacterium, a fungus, a virus and an aquatic animal, wherein said aquatic animal comprises a fish, a shellfish, and a crustacean. [00248] In some embodiments, the target is agricultural pests such as mites, aphids, whiteflies and thrips among the agricultural pests. Examples of other agricultural insect pests than the mites, aphids, whiteflies and thrips include diamondback moth (Plutella xylostella), cabbage armyworm (Mamestra brassicae), common cutworm (Spodoptera litura), codlingmoth (Cydia pomonella), bollworm (Heliothis zea), tobacco budworm (Heliothis virescens), gypsy moth (Lymantria dispar), rice leafroller (Cnaphalocrocis medinalis), smaller tea tortrix 62 303137700 (Adoxophyes sp.), Colorado potato beetle (Leptinotarsa decemlineata), cucurbit leaf beetle (Aulacophora femoralis), boll weevil (Anthonomus grandis), planthoppers, leafhoppers, scales, bugs, grasshoppers, anthomyiid flies, scarabs, black cutworm (Agrotis ipsilon), cutworm (Agrotis segetum) and ants. [00249] In addition, examples of other agricultural pests include soil pests, such as plant parasitic nematodes such as root-knot nematodes (Meloidogynidae), cyst nematodes (Heteroderidae), root-lesion nematodes (Pratylenchidae), white-tip nematode (Aphelenchoi desbesseyi), strawberry bud nematode (Nothotylenchus acris) and pine wood nematode (Bursaphelenchus xylophilus); gastropods such as slugs and snails; and isopods such as pill bugs (Armadillidium vulgare) and pill bugs (Porcellio scaber). [00250] Examples of other insect pests include hygienic insect pests such as tropical rat mite (Ornithonyssus bacoti), cockroaches, housefly (Musca domestica) and house mosquito (Culex pipiens pallens); stored grain insect such as angoumois grain moth (Sitotroga cerealella), adzuki bean weevil (Callosobruchus chinensis), red flour beetle (Tribolium castaneum) and mealworms; clothes insect pests such as casemaking clothes moth (Tinea translucens) and black carpet beetle (Attagenus unicolor japonicus); house and household insect pests such as subterranean termites; domestic mites such a mold mite (Tyrophaqus putrescentiae), Dermatophagoides farinae and Chelacaropsis moorei; and hygienic insect pests such as tropical rat mite (Ornithonyssus bacoti). [00251] Insect pests include insects selected from the orders Coleoptera, Diptera, Hymenoptera, Lepidoptera, Mallophaga, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Isoptera, Anoplura, Siphonaptera, Trichoptera, etc. [00252] In some embodiments, the insects are selected from cotton bollworm, native budworm, green mirids, aphids, green vegetable bugs, apple dimpling bugs, thrips (plaque thrips, tobacco thrips, onion thrips, western flower thrips), white flies and two spotted mites. In an embodiment the insect pests of animals include fleas, lice, mosquitoes, flies, tsetse flies, ants, ticks, mites, silverfish and chiggers. The above agricultural pests and insect pests are described, for example, in U.S. Patent Application Nos. 2012/0016022 and 2016/0174571, which are incorporated by reference herein in their entirety. Delivery Amounts [00253] In some embodiments, biologically active compounds are encapsulated within the minicells described herein and delivered to a desired target. Amounts of an biologically active compound of interest are provided herein with percent weight proportions of the various 63 303137700 components used in the preparation of the minicell for the encapsulation and deliver of biologically active compounds. [00254] The percent weight proportions of the various components used in the preparation of the minicell for the encapsulation and deliver of biologically active compounds can be varied as required to achieve optimal results. In some embodiments, the biologically active compounds including, but are not limited to a nucleic acid, a polypeptide, a metabolite, a semiochemical and a micronutrient polypeptide, are present in an amount of about 0.1 to about 90% by weight, is present in an amount of about 0.5 to about 80% by weight, 1 to about 70% by weight, 2 to about 60% by weight, 3 to about 55% by weight, 5 to about 50% by weight, 10 to about 45% by weight, and 15 to about 40% by weight, based on the total weight of the minicell within which an active compound of interest is encapsulated. When a polymer is used in the preparation of the minicell disclosed herein, according to one embodiment it is present in an amount of about 0.01 to about 10% by weight based on the total weight of the minicell disclosed herein. When a co-solvent is used in the preparation of the minicell disclosed herein, according to one embodiment it is present in an amount of about 0.1 to about 30% by weight based on the total weight of the minicell disclosed herein. Alternate percent weight proportions are also envisioned. For example, the biologically active compound of interest can be present in an amount of up to about 50% by weight; the solvent can be present in an amount of up to about 70% by weight; the surfactant can be present in an amount of up to about 40% by weight and the water can be present in an amount of from about 1 to about 90% by weight, based on the total weight of the minicell disclosed herein. [00255] Among the various aspects of the present disclosure is a minicell in the form of encapsulation of an biologically active compound of interest at least about 0.1%, about 0.2%, about 0.3%, about 0.4%, about 0.5%, about 0.6%, about 0.7%, about 0.8%, about 0.9%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%, by weight of the biologically active compound within the minicell. [00256] In other embodiments, the biologically active compound within the minicell is present in an amount of at least about 0.01, about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.1, about 0.2, about 0.3, about 0.4, about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 64 303137700 16, about 17, about 18, about 19, about 20, about 30, about 40, about 50, about 60, about 70, about 80, about 90, or about 100 g/L. [00257] In another embodiment, the biologically active compound of interest and the minicell are present in compositions of the disclosure in a weight ratio of at least 1:200, 1:195, 1:190, 1:185, 1:180, 1:175, 1:170, 1:165, 1:160, 1:155, 1:150, 1:145, 1:140, 1:135, 1:130, 1:125, 1:120, 1:115, 1:110, 1:105, 1:100, 1:95, 1:90, 1:85, 1:80, 1:75, 1:70, 1:65, 1:60, 1:55, 1:50, 1:45, 1:40, 1:35, 1:30, 1:25, 1:20, 1:15, 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2, 1:1, 2:1, 3:1, 4:1, 5:1, 6:1,7:1, 8:1, 9:1, 10:1, 15:1, 20:1, 25:1, 30:1, 35:1, 40:1, 45:1, 50:1, 55:1, 60:1, 65:1, 70:1, 75:1, 80:1, 85:1, 90:1, 95:1, 100:1, 110:1, 120:1, 130:1, 140:1, 150:1, 160:1, 170:1, 180:1, 190:1 or 200:1. In another embodiment, the biologically active compound of interest and the minicell are present in a weight ratio of from about 1:50 to about 50:1, from about 1:40 to about 40:1, from about 1:30 to about 30:1, from about 1:20 to about 20:1, from about 1:10 to about 10:1, or from about 1:5 to about 5:1. [00258] In further embodiments, the density of the formulation of the minicell encapsulating the biologically active compound is least 0.01, at least 0.02, at least 0.03, at least 0.04, at least 0.05, at least 0.06, at least 0.07, at least 0.08, at least 0.09, at least 0.1, at least about 0.2, at least about 0.3, at least about 0.4, at least about 0.5, at least about 0.6, at least about 0.7, at least about 0.8, at least about 0.9, at least about 1.0, at least 1.1, at least about 1.2, at least about 1.3, at least about 1.4, at least about 1.5, at least about 1.6, at least about 1.7, at least about 1.8, at least about 1.9, at least about 2.0, at least 2.1, at least about 2.2, at least about 2.3, at least about 2.4, at least about 2.5, at least about 2.6, at least about 2.7, at least about 2.8, at least about 2.9, at least about 3.0, at least 3.1, at least about 3.2, at least about 3.3, at least about 3.4, at least about 3.5, at least about 3.6, at least about 3.7, at least about 3.8, at least about 3.9, at least about 4.0, at least 4.1, at least about 4.2, at least about 4.3, at least about 4.4, at least about 4.5, at least about 4.6, at least about 4.7, at least about 4.8, at least about 4.9, at least about 5.0, at least about 5.5, at least about 6.0, at least about 6.5, at least about 7.0, at least about 7.5, at least about 8.0, at least about 8.5, at least about 9.0, at least about 9.5, or at least about 10.0 grams/liter. [00259] In some embodiments, an biologically active compound of interest, for example, is present in at least about 20% of the total mass of the formulated product. In further embodiments, about 20 to 40% of the total mass of the formulated product is provided for the biologically active compound disclosed herein and the remaining about 60 to 80% of the mass is from the minicell. 65 303137700 [00260] In some embodiments, more than one non-expressed biologically active compounds can be encapsulated within the minicell. In another embodiment, the formulated product comprises two biologically active compounds that are present in compositions of the disclosure in a weight ratio of at least 1:10, 1:9, 1:8, 1:7, 1:6, 1:5, 1:4, 1:3, 1:2,1:1, 2:1, 3:1, 4:1, 5:1, 6:1,7:1, 8:1, 9:1, or 10:1. [00261] In terms of amounts of the biologically active compound, about a concentration of about 0.01-20, about 0.1-15, about 0.2-10, about 0.3-9, about 0.3-8, about 0.5-5, about 1-3 g/L is provided for the formulated product. [00262] In some embodiments, the targeted delivery and controlled release disclosed herein can improve efficacy of the biologically active compounds so that the amounts of the biologically active compound can be used less. The formulation of the minicell can be in a liquid or solid form. In some embodiments, the formulated product is a liquid form such as a solution. In some embodiments, the formulated product is a solid form such as a powder. [00263] In some embodiments, the agricultural formulation further comprises an agricultural chemical that is useful for promoting plant growth, reducing weeds, or reducing pests. In some embodiments, the agricultural formulation further comprises at least one of a fungicide, an herbicide, a pesticide, a nematicide, an insecticide, a plant activator, a synergist, an herbicide safener, a plant growth regulator, an insect repellant, an acaricide, an algaecide, a bactericide, a virucide, an ovicide, a rodenticide, a larvicide, a molluscicide, a pediculicide, or a fertilizer. In some embodiments, the agricultural formulation further comprises a surfactant. In some embodiments, the agricultural composition further comprises a carrier. The present disclosure provides for agricultural compositions formulated for contacting to pests and/or plants. [00264] The formulations can be suitable for treating plants or plant propagation material, such as seeds, in accordance with the present disclosure, e.g., in a carrier. Suitable additives include buffering agents, wetting agents, coating agents, polysaccharides, and abrading agents. Exemplary carriers include water, aqueous solutions, slurries, solids and dry powders (e.g., peat, wheat, bran, vermiculite, clay, pasteurized soil, many forms of calcium carbonate, dolomite, various grades of gypsum, bentonite and other clay minerals, rock phosphates and other phosphorous compounds, titanium dioxide, humus, talc, alginate and activated charcoal. Any agriculturally suitable carrier known to one skilled in the art would be acceptable and is contemplated for use in the present invention). Optionally, the formulations can also include at least one surfactant, herbicide, fungicide, pesticide, or fertilizer. [00265] In some embodiments, the agricultural composition further comprises at least one of a surfactant, an herbicide, a pesticide, such as but not limited to a fungicide, a bactericide, an 66 303137700 insecticide, an acaricide, and a nematicide, a plant activator, a synergist, an herbicide safener, a plant growth regulator, an insect repellant, or a fertilizer. Pesticides [00266] Pesticides are substances that are meant to control pests. As used herein, “pesticide” includes all of the following: herbicide, insecticides (which may include insect growth regulators, termiticides, etc.), nematicide, molluscicide, piscicide, avicide, rodenticide, bactericide, insect repellent, animal repellent, antimicrobial, and fungicide. Most pesticides are intended to serve as plant protection products (also known as crop protection products), which in general, protect plants from weeds, fungi, or insects. [00267] In general, a pesticide is a chemical or biological agent that deters, incapacitates, kills, or otherwise discourages pests. Target pests can include insects, plant pathogens, weeds, molluscas, lices, birds, mammals, fish, nematodes (roundworms), and microbes that destroy property, cause nuisance, or spread disease, or are disease vectors. In some embodiments, one or more biological pesticide, includes but not limited to, Pasteuria spp., Paeciliomyces, Pochonia chlamydosporia, Myrothecium metabolites, Muscodor volatiles, Tagetes spp., bacillus firmus, including bacillus firmus CNCM 1-1582. Insecticides [00268] Insecticides are substances used to kill insects and some other arthropods (mites, ticks, spiders, etc.) or to prevent them from causing damage. Insecticides include ovicides and larvicides used against insect eggs and larvae, respectively. Insecticides can be classified into two major groups: systemic insecticides, which have residual or long term activity; and contact insecticides, which have no residual activity. [00269] The mode of action describes how the pesticide kills or inactivates a pest. Many insecticides act at specific sites in the insect's nervous system. These usually provide very quick knockdown of insects that may ultimately die from dehydration or starvation. The insecticides usually are sprayed on infested plants or surface on which they rest. Depending on the pest, the insecticide may kill by direct contact with the spray droplets, ingestion of treated foliage, or prolonged contact with the residue on a treated surface. [00270] In some aspects, there are some types of insecticides, but not limited to; (i) organochlorides, such as DDT, (ii) cholinesterase inhibitors that interfere with nerve impulse transmission at the synapse gap, such as organophosphate (malathion, diazinon, chlorpyrifos, parathion, acephate) and carbamate (carbaryl, propoxur, methomyl) insecticides; (iii) 67 303137700 pyrethroids (permethrin, cypermethrin, deltamethrin, bifenthrin) as nonpersistent sodium channel modulators; (iv) neonicothinoids as acetylcholine receptor agonists (such as imidacloprid, clothianidin, thiamethoxam, acetamiprid, thiacloprid); (v) butenolides as acetylcholine receptor agonist (such as flupyradifurone); (vi) ryanoids (such as chlorantraniliprole); (vii) bacterial toxins derived from soil microorganisms, such as Bacillus thuringiensis (Bts) and spinosyns (spinosad, spinetoram); (viii) botanical insecticides extracted from plants, such as pyrethrins, nicotine, and azadirachtin; (ix) insect growth regulators (IGRs) based on hormones that regulate arthropod development (such as benzoylphenyl ureas, diflubenzuron, methoprene, hydroprene, kinoprene, methoprene, pyriproxyfen, fenoxycarb, and tebufenozide), (x) diamides (chlorantraniliprole, flubendiamide, cyantraniliprole), (xi) Bacillus thuringiensis (Bt) toxin (Bt kurstaki, Bt israelensis, Bt tenebrionis), (xii) biopesticides (azadirachtin from Neem oil; Beauveria bassiana (fungal insecticide), and Spinosad derived from a soil bacterium, and (xiii) others (chlorfenapyr, cyfluthrin, indoxacarb, fipronil). [00271] The present disclosure teaches that RNAi can be utilized to alter and/or regulate (either downregulate or upregulate) expression of genes associated with or responsible for insecticide resistance. As one example, the amino acid mutations in ryanodine receptor (RyR) and elevated activity of detoxification enzymes have been associated with the diamide insecticide resistance in the diamondback moth, Plutella xylostella (L.). Higher expression of P. xylostella RyR (PxRyR) is detected in field collected resistant populations, compared to that in a susceptible population. Suppression of P. xylostella RyR mRNA (PxRyR) using RNAi (such as micro RNAs; miRNAs) can restore the toxicity of chlorantraniliprole against the fourth instar larvae from the resistant diamondback moth population (Li, X et al., Sci Rep 5, 14095 (2015). The expression of PxRyR can be regulated by two miRNAs, miR-7a and miR-8519 in P. xylostella. As the target of diacylhydrazines insecticides, the expression of ecdysone receptor (EcR) is regulated by miRNA-281 in the silkworm, Bombyx mori (Jiang et al, Insect Biochem Molec 43:692-700, 2013). The reduced expression of nicotinic acetylcholine receptor (nAChR) VXEXQLW^ Į^^ LV^ UHVSRQVLEOH^ IRU^ WKH^ LPLGDFORSULG^ UHVLVWDQFH^ LQ^ KRusefly, Musca domestica (Markussen et al, Pest Manag Sci 66:1257-1262, 2010). [00272] Also, the knockdown of a cytochrome P450 gene, CYP6BG1, from the diamondback moth by RNAi imposes a significant reduction in larval resistance to permethrin in the diamondback moth (Bautista MA, et al., Insect Biochem Mol Biol. 2009 Jan;39(1):38-46). [00273] It is reported by Tabashnik BE et al. (Proc Natl Acad Sci U S A. 1997;94(5):1640- 1644) that one gene in diamondback moth confers resistance to four Bacillus thuringiensisௗWR[LQV^^,Q^Vome embodiments, the agricultural compositions/formulations of the 68 303137700 present disclosure using the minicell platform are used to knockdown expression of the gene conferring resistance to four Bt toxins by RNAi. [00274] Insects including fall armyworm, can develop resistance to many chemical insecticides: chlorpyriphos, permethrin, flubendamide, chlorantraniliprole, methomyl, thiodicarb, permethrin, chlorpyriphos, zeta-cypermethrin, deltamethrin, triflumuron, spinetoram, spinosad, emamectin benzoate and abamectin. [00275] In some embodiments, the minicell technology for delivery of RNAi molecules to a target with insecticides can be used to manage resistance of the fall armyworm to chemical insecticides as well as transgenic Bt corns. [00276] In some embodiments, Monera et al. (J Econ Entomol actions 2019 Mar 21;112(2):792-802) reports that the resistance levels of fall armyworm to insecticides of different modes of action in fall armyworm populations from Puerto Rico and several Mexican states with different insecticide use patterns. Mexican populations that expressed higher resistance ratios (RR50) were: Sonora (20-fold to chlorpyriphos), Oaxaca (19-fold to permethrin), and Sinaloa (10-fold to flubendamide). The Puerto Rico population exhibited a remarkable field-evolved resistance to many pesticides. The RR50 to the insecticides tested were: flubendiamide (500-fold), chlorantraniliprole (160-fold), methomyl (223-fold), thiodicarb (124-fold), permethrin (48-fold), chlorpyriphos (47-fold), zeta-cypermethrin (35- fold), deltamethrin (25-fold), triflumuron (20-fold), spinetoram (14-fold). Spinosad (eightfold), emamectin benzoate and abamectin (sevenfold) displayed lower resistance ratio. These compounds are still effective to manage fall armyworm resistance in Puerto Rico. Fall armyworm populations from Mexico show different levels of susceptibility, which may reflect the heterogeneity of the pest control patterns in this country. [00277] In some embodiments, exemplary insecticides, include, but are not limited to, thiamethoxam, imidacloprid, clothianidin, lamda-cyhalothrin, tefluthrin, beta-cyfluthrin, permethrin, abamectin, fipronil, cyanotraniliprole, chlorantraniliprole, and spinosad. Details (e.g., structure, chemical name, commercial names, etc.) of each of the above insecticides with a common name can be found in the e-Pesticide Manual, version 3.1, 13th Edition, Ed. CDC Tomlin, British Crop Protection Council, 2004-05. The above compounds are described, for example, in U.S. Pat. No. 8,124,565, which is incorporated by reference herein in its entirety. Fungicides [00278] Fungicides are biocidal chemical compounds or biological organisms used to kill parasitic fungi or their spores. They may be applied to seeds, soil, or foliage. Fungi can cause 69 303137700 serious damage in agriculture, resulting in critical losses of yield, quality, and profit. Fungicides are used both in agriculture and to fight fungal infections in animals. Chemicals used to control oomycetes, which are not fungi, are also referred to as fungicides, as oomycetes use the same mechanisms as fungi to infect plants. [00279] Exemplary lists of fungicides, but not limited to, are as follows: Azoxystrobin, Boscalid, BYF 14182, Carbendazim, Carboxin, Chlorothalonil, Fenamidone, Fludioxonil, Fluopicolide, Fluoxastrobin, Fluquinconazole, Flutriafol, Ipconazole, Iprodione, Isotianil, Mancozeb, Mefenoxam, Metalaxyl, Myclobutanil, Pencycuron, Prochloraz, Propiconazole, Prothioconazole, Pyraclostrobin, Pyrimethanil, Silthiopham, Tebuconazole, Thiophanate- methyl, Thiram, Tolylfluanid, Triadimenol, Triazoxide, Trifloxystrobin, Triflumuron, Triticonazole. [00280] In some embodiments, additional exemplary fungicides include, but are not limited to, sedaxane, fludioxonil, penthiopyrad, prothioconazole, flutriafol, difenoconazole, azoxystrobin, captan, cyproconazole, cyprodinil, boscalid, diniconazole, epoxiconazole, fluoxastrobin, trifloxystrobin, metalaxyl, metalaxyl-M (mefenoxam), fluquinconazole, fenarimol, nuarimol, pyrifenox, pyraclostrobin, thiabendazole, tebuconazole, triadimenol, benalaxyl, benalaxyl-M, benomyl, carbendazim, carboxin, flutolanil, fuberizadole, guazatine, myclobutanil, tetraconazole, imazalil, metconazole, bitertanol, cymoxanil, ipconazole, iprodione, prochloraz, pencycuron, propamocarb, silthiofam, thiram, triazoxide, triticonazole, tolylfluanid, isopyrazam, mandipropamid, thiabendazole, fluxapyroxad, and a manganese compound (such as mancozeb, maneb). [00281] In some embodiments, further exemplary fungicides include, but are not limited to, Cyprodinil ((4-cyclopropyl-6-methyl-pyrimidin-2-yl)-phenyl-amine), Dodine, Chlorothalonil, Folpet, Prothioconazole, Boscalid, Proquinazid, Dithianon, Fluazinam, Ipconazole, and Metrafenone. Some of the above compounds are described, for example, in “The Pesticide Manual” [The Pesticide Manual—A World Compendium; Thirteenth Edition; Editor: C. D. S. Tomlin; The British Crop Protection Council, 2003]. The above compounds are described, for example, in U.S. Pat. No. 8,349,345, which is incorporated by reference herein in its entirety. [00282] In some embodiments, other exemplary fungicides includes, but are not limited to, fludioxonil, metalaxyl and a strobilurin fungicide, or a mixture thereof. In some embodiments, the strobilurin fungicide is azoxystrobin, picoxystrobin, kresoxim-methyl, or trifloxystorbin. In some embodiments, the agricultural chemical formulation comprises an effective amount of one or more of an insecticide selected from a phenylpyrazole and a neonicotinoid. In some embodiments, the phenylpyrazole is fipronil and the neonicotinoid is selected from 70 303137700 thiamethoxam, imidacloprid, thiacloprid, clothianidin, nitenpyram and acetamiprid. The above compounds are described, for example, in U.S. Pat. No. 7,071,188, which is incorporated by reference herein in its entirety. [00283] Botrytis cinerea is a major plant pathogen, causing gray mold rot in a variety of cultures. Repeated fungicide applications can result in the development of fungal populations with resistance to one or more fungicides. It is reported that fungicide resistance frequencies and the occurrence of multiple resistance in Botrytis cinerea strains were found from raspberries, strawberries, grapes, stone fruits and ornamental flowers treated with repeated fungicide treatments. B. cinerea strains carrying multiple resistance mutations against all classes of site-specific fungicides were detected with the chemical control of Borytis using quinone-outside inhibitors (QoI) such as azoxystrobin, trifloxystrobin or pyraclostrobin; succinate dehydrogenase inhibitors (SDHI) such as boscalid or the recently registered fluopyram; anilinopyrimidines (e.g., cyprodinil, pyrimethanil, mepanipyrim); phenylpyrroles (fludioxonil); and hydroxyanilides (fenhexamid) (Rupp S et al., Front Microbiol. 2017 Jan 3;7:2075). There is an alarming increase in the occurrence of multiresistance in B. cinerea populations from different cultures, which presents a major threat to the chemical control of gray mold. [00284] The present disclosure teaches that RNAi can be utilized to negatively regulate expression of genes associated with fungicide resistance. As one example, azoles are one of the main treatments in the management of Aspergillus diseases caused by ubiquitous fungi, such as Aspergillus fumigatus. The main resistance mechanism is a combination of alterations in the gene cyp51A (TR34/L98H) (Berger S et al., Front Microbiol. 2017 Jun 7;8:1024). In some embodiments, the cyp51A gene can be a target of the minicell-based RNAi along with fungicide treatment including azoles. Herbicides [00285] Herbicides, also known as weedkillers, are substances used to control undesired plants. Non-selective herbicides (sometimes called total weedkillers in commercial products) can be used to clear ground and sites where all plant material are killed with contact of the herbicides. A selective herbicide controls or suppresses unwanted plant species without seriously affecting the growth of desirable plant species. Selective herbicides are used to kill weeds without harming nearby desirable plants. For example, 2,4-D may be used for selective control of many broadleaf weeds without significant injury to desirable grasses. Herbicides are often synthetic mimics of natural plant hormones which interfere with growth of the target plants. 71 303137700 [00286] Due to herbicide resistance, a number of products combine herbicides with different means of action. Integrated pest management may use herbicides alongside other pest control methods. However, these approaches have led to the evolution and widespread distribution of herbicide-resistant weeds, which has become a challenge for crop producers and land managers After commercialization of glyphosate-tolerant soybean in 1996 and corn in 1997, glyphosate has been used extensively for weed control and resulted in the evolution of glyphosate-resistant weeds. Even the weed species with multiple-resistance has acquired resistance against herbicides belonging to different chemistries. This is due to overreliance or continuous use of two or more selected herbicides over time. Multiple-resistance has been reported in several weed species including Palmer amaranth (Amaranthus palmeri). [00287] In some embodiments, exemplary herbicides includes, but are not limited to, paraquat, mesotrione, sulcotrione, clomazone, fentrazamide, mefenacet, oxaziclomefone, indanofan, glyphosate, prosulfocarb, molinate, triasulfuron, halosulfuron-methyl, pretilachlor, topramezone, tembotrione, isoxaflutole, fomesafen, clodinafop-propargyl, fluazifop-P-butyl, dicamba, 2,4-D (2,4-Dichlorophenoxyacetic acid), dicamba, atrazine, paraquat, S-metolachlor, glufosinate, fluroxypyr, pinoxaden, bicyclopyrone, metolachlor, and pyroxasulfone. The above herbicidal active ingredients are described, for example, in “The Pesticide Manual”, Editor C. D. S. Tomlin, 12th Edition, British Crop Protection Council, 2000, under the entry numbers added in parentheses; for example, mesotrione (500) is described therein. The above compounds are described, for example, in U.S. Pat. No. 7,338,920, which is incorporated by reference herein in its entirety. [00288] In some embodiments, the minicell-based RNAi along with herbicide treatment are taught herein. RNA interference (RNAi) [00289] RNA interference (RNAi) is a biological process that regulates the expression of genes in cells. It involves the silencing of specific genes through the use of small RNA molecules. These small RNAs, typically around 20-25 nucleotides in length, can interfere with the translation of messenger RNA (mRNA) or target mRNA for degradation, thereby preventing the production of the corresponding protein. [00290] The process of RNA interference involves with the introduction of double-stranded RNA (dsRNA) into the cell. This dsRNA is cleaved by an enzyme called Dicer into small interfering RNAs (siRNAs) or microRNAs (miRNAs). These small RNAs then bind to a protein complex called the RNA-induced silencing complex (RISC). Within the RISC, one of 72 303137700 the strands of the siRNA or miRNA, known as the guide strand, directs the complex to complementary sequences on target mRNA molecules. The RISC then either cleaves the mRNA, preventing its translation into protein, or inhibits translation by other means. [00291] The present disclosure teaches that RNAi is used as a promising tool for pest control, offering a targeted and environmentally friendly approach to managing insect pests. In some embodiments, RNAi is utilized as a biopesticide by identify specific genes essential for the survival or development of the target insect pest. These genes could be involved in vital physiological processes such as metabolism, development, or reproduction. Once the target genes are identified, double-stranded RNA (dsRNA) molecules are designed to match sequences within these genes. These dsRNAs can be synthesized in the laboratory. Then, the dsRNA molecules are encapsulated and effectively delivered into the target pests. This delivery can be achieved through various delivery methods such as spraying, application as powder or granules. Upon exposure to the dsRNA, the insect pests ingest or absorb it. Inside the insect cells, the dsRNA is processed into small interfering RNAs (siRNAs) by enzymes such as Dicer. The siRNAs are incorporated into the RNA-induced silencing complex (RISC), where they guide the complex to target mRNA molecules that have complementary sequences. The RISC then cleaves the target mRNA, leading to its degradation or inhibition of translation. By targeting essential genes, RNAi disrupts vital physiological processes in the insect pests, leading to reduced survival, development, or reproduction. This effectively controls pest populations while minimizing off-target effects on non-target organisms and the environment. [00292] The present disclosure teaches that RNAi-based biopesticides offer advantages over traditional chemical pesticides. They are highly specific, targeting only the desired insect pests while leaving beneficial organisms unharmed. Additionally, they are environmentally friendly, as they degrade rapidly in the environment and reduce the risk of pesticide residues in food and water. [00293] In some embodiments, a dsRNA can target more than one target gene if the dsRNA sequence is processed into small RNAs that can recognize and/or bind to more than one target gene transcript based on its sequence homology. In some embodiments, a dsRNA can target 1 target gene, 2 target genes, 3 target genes, 4 target genes, 5 target genes, 6 target genes, 7 target genes, 8 target genes, 9 target genes, 10 target genes, or more than 10 target genes for degradation of target gene transcripts and/or inhibition of translation of target gene transcripts. [00294] The present disclosure teaches that minicell encapsulates dsRNA that will be process into small RNAs or the processed small RNAs for RNAi in order to regulate the expression of target genes in cells. The present disclosure teaches benefits of minicell-mediated dsRNAs 73 303137700 (which is interchangeably used with minicell-encapsulated dsRNA) that confers protection and stabilization of dsRNAs by a barrier (that is, membranes of minicells). In some embodiments, minicells provide a protective barrier around the biological ingredients/compounds or chemical compounds (such as agrochemicals), shielding them from degradation due to environmental factors such as UV radiation, temperature fluctuations, moisture, and microbial activity. This helps maintain the viability and efficacy of the biologicals or chemicals during storage and application. [00295] The present disclosure teaches that minicells can provide targeted delivery. In some embodiments, minicell-encapsulated biologicals (such as dsRNA or small RNA) and/or minicell-encapsulated agrochemicals can be designed to release their active ingredients slowly and steadily over time, allowing for controlled and targeted delivery to specific sites, such as pests, insects, plants or soil zones where they are needed most. This ensures optimal utilization of the biologicals and/or chemicals and reduces wastage. [00296] The present disclosure teaches that minicells can provide enhanced persistence. In some embodiments, minicells help prolong the persistence of biologicals or agrochemicals in the environment by preventing rapid degradation or wash-off. This extends their effectiveness and reduces the frequency of application, leading to cost savings and environmental benefits. [00297] The present disclosure teaches that minicells can provide improved handling and application. Minicell formulations can be easier to handle and apply compared to their liquid or powder counterparts. They can be formulated into various forms such as granules, pellets, or coated seeds, which are convenient to transport, store, and apply using standard agricultural equipment. [00298] In some embodiments, a minicell can encapsulate 1 dsRNA, 2 dsRNAs, 3 dsRNAs, 4 dsRNAs, 5 dsRNAs, 6 dsRNAs, 7 dsRNAs, 8 dsRNAs, 9 dsRNAs, 10 dsRNAs, or more than 10 dsRNAs, each of which targets a different target gene transcript. Resistance potential in SDHI fungicides [00299] SDHI (succinate dehydrogenase inhibitor) fungicides are a class of fungicides that target the succinate dehydrogenase enzyme complex in fungi, disrupting cellular respiration and leading to fungal death. Here are some examples of SDHI fungicides: Boscalid, Flutolanil, Isopyrazam, Bixafen, Fluopyram, and Isofetamid. [00300] Concerns regarding pathogen resistance to SDHI fungicides, like other site-specific penetrant fungicides, have emerged due to reduced sensitivity across various cropping systems. Instances of reduced sensitivity to SDHI fungicides have been reported in pathogens such as 74 303137700 Botrytis cinerea in fruit crops (Yin et al.2011), Alternaria spp. in nut and potato crops (Avenot et al, 2008; Gudmestad et al, 2013), and Didymella bryoniae in cucurbits (Avenot et al, 2011). Also, resistance of the dollar spot pathogen (Clarireedia spp.) to certain SDHI fungicides is also known. [00301] Understanding the issue of cross-resistance among different chemical groups and active ingredients of SDHI fungicides is complex. Isolates resistant to certain SDHI fungicides remaining sensitive to others. For example, some isolates of Didymella bryoniae resistant to boscalid and penthiopyrad were found to be sensitive to fluopyram. Similarly, in cases of boscalid control failures with early blight of potato, isolates resistant to boscalid remained sensitive to fluopyram. Additionally, isolates resistant to boscalid were found to be sensitive to penthiopyrad. Prototoxins [00302] Protein toxins, also known as protoxins produced by bacteria or plants represent potent cytotoxic agents that may be coupled to specific carrier ligands used for cellular targeting. Current targeted toxins are comprised of fusion proteins that contain a potent toxin engineered in bacteria, along with a carrier ligand. The mode of action of protein toxins involves disrupting essential biological processes in pests. Exemplary protoxins includes, but are not limited to, (i) insecticides, which are neonicotinoids, pyrethroids, Cry and Cyt toxins derived from Bacillus thuringiensis (Bt); (ii) fungicides, which are SDHI fungicides (Succinate Dehydrogenase Inhibitors), triazoles, strobilurins; (iii) herbicides, which are glyphosate, ALS inhibitors (Acetolactate Synthase Inhibitors), and photosystem II inhibitors (such as atrazine and diuron). [00303] Bacillus thuringiensis toxin (Bt toxin) refers to a group of protein toxins produced by the bacterium Bacillus thuringiensis (Bt). These toxins are commonly used as biopesticides to control insect pests in agriculture and forestry. Bt toxins are insecticidal proteins that are selectively toxic to certain insect species while being harmless to humans, animals, and most beneficial insects. They function by disrupting the gut lining of susceptible insect larvae, leading to paralysis, starvation, and eventual death. There are several types of Bt toxins, each targeting specific groups of insect pests. Exemplary types of Bt toxins used in pest control include, but are not limited to, Cry toxins as pore-forming toxins that create pores in the gut epithelial cells of susceptible insects (Cry1A, Cry2A, Cry3, Cry4, Cry5, and others), .Cyt toxins as cytolytic toxins that disrupt the cell membranes of insect gut cells and often act synergistically with Cry toxins to enhance insecticidal activity, and Vip toxins (Vegetative 75 303137700 insecticidal protein toxins) that target the midgut epithelium of susceptible insects and disrupt cellular functions. [00304] In biopesticides, Bt toxins can be applied directly to crops as sprays or introduced into crop plants through genetic engineering to confer resistance to specific insect pests. Bt-based biopesticides are widely used in organic and conventional agriculture as environmentally friendly alternatives to synthetic chemical insecticides. Use of minicell technology [00305] In some embodiments, the disclosure provides a plurality of minicells. In some embodiments, the disclosure provides a plurality of minicells comprising at least one biologically active compound within said minicells. In other embodiments, the disclosure provides a plurality of minicells comprising at least one biocontrol within said minicells. [00306] In some embodiments, the disclosure provides a plurality of minicells comprising at least one nucleic acid such as RNA molecule including antisense nucleic acid, dsRNA, shRNA, siRNA, miRNA, ribozyme, and aptamer within said minicells. [00307] In some embodiments, the disclosure provides a plurality of minicells comprising at least one pesticide selected from herbicide, insecticides (which may include insect growth regulators, termiticides, etc.), a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, an avicide, and combinations thereof. In some embodiments, the disclosure provides a plurality of minicells comprising at least one herbicide. In some embodiments, the disclosure provides a plurality of minicells comprising at least one fungicide. In some embodiments, the disclosure provides a plurality of minicells comprising at least one antifungal. In some embodiments, the disclosure provides a plurality of minicells comprising at least one bactericide. In some embodiments, the disclosure provides a plurality of minicells comprising at least one pesticides within said minicells. In some embodiments, the disclosure provides a plurality of minicells comprising at least one pesticides within said minicells. In some embodiments, the disclosure provides a plurality of minicells comprising at least one pesticides within said minicells. [00308] In some embodiments, the disclosure provides a composition comprising a plurality of intact, bacterially-derived minicells. In some embodiments, the disclosure provides a composition comprising a plurality of intact, bacterially-derived minicells comprising at least one biologically active compound within said minicells. 76 303137700 [00309] The present disclosure provides an agricultural composition, comprising a minicell encapsulating a nucleic acid that is capable of inducing RNA interference in an agricultural pest. In some embodiments, the nucleic acid reduces resistance to or tolerance of the pesticide in the agricultural pest. In some embodiments, the agricultural pest is resistant to or tolerant of the pesticide. In some embodiments, the pesticide is a chemical pesticide or a biological pesticide, which is an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, or an avicide. In some embodiments, the agricultural pest is insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, or bird. [00310] In some embodiments, the nucleic acid is capable of recovering the agricultural pest’s sensitivity or susceptibility to the pesticide. In some embodiments, the nucleic acid is capable of altering expression of a gene responsible for pesticide resistance or tolerance. In some embodiments, the expression of the gene responsible for pesticide resistance or tolerance is downregulated. In other embodiments, the expression of the gene responsible for pesticide resistance or tolerance is upregulated. [00311] In some embodiments, the gene responsible for pesticide resistance or tolerance is an ion channel gene, a detoxification gene, a target site resistance gene, or a transporter gene. In some embodiments, the ion channel gene is a gene encoding Ryanodine receptor (RyR) or Voltage-gated sodium channel (VGSC). In some embodiments, the detoxification gene is a gene encoding UDP-glycosyltransferase (UGT), Cytochrome P450 monooxygenase, Esterase, Carboxylesterase (CarE), or Glutathione S-transferase (GST). In some embodiments, the target site resistance gene is a gene encoding Acetylcholinesterase (AChE), Voltage-gated sodium channel (VGSC), Gamma-aminobutyric acid (GABA) receptor, Nicotinic acetylcholine receptor (nAChR), or Glutamate-gated chloride channel (GluCl). In some embodiments, the transporter gene is a gene encoding ATP-binding cassette (ABC) transporter, Solute carrier (SLC) transporter, Major facilitator superfamily (MFS) transporter, or P-glycoprotein. [00312] In some embodiments, the nucleic acid is capable of inducing RNA interference in at least one member from an order selected from the group consisting of: Coleoptera, Diptera, Hymenoptera, Lepidoptera, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Siphonaptera, and Trichoptera. In some embodiments, the nucleic acid is capable of inducing RNA interference in a member of the order Lepidoptera. In some embodiments, the nucleic acid is capable of inducing RNA interference in a member of the genus Plutella. In some 77 303137700 embodiments, the nucleic acid is capable of inducing RNA interference in a member of the genus Spodoptera. [00313] In some embodiments, the nucleic acid is a RNA molecule, which is a double-stranded RNA (dsRNA), a short-hairpin RNA (shRNA), a small-interfering RNA (siRNA), a microRNA (miRNA), or an antisense RNA,. [00314] In some embodiments, the minicell is ribonuclease deficient. In some embodiments, the minicell comprises at least one fusion protein. In some embodiments, the minicell comprises at least one fusion protein expressed on the surface of the minicell. In some embodiments, the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising at least one target cell adhesion moiety. In some embodiments, the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising a carbohydrate binding molecule. [00315] The present disclosure provides the agricultural composition comprises a minicell- encapsulated dsRNA (ME-dsRNA) and an agricultural chemical or biological pesticide. That is, the minicell-mediated RNA molecule can be co-applied with an agricultural chemical or biological pesticide. [00316] In some embodiments, the pesticide can be applied exogenously along with the minicell-mediated RNA molecule. In other embodiments, the pesticide is encapsulated by another second minicell and applied with a first minicell encapsulating the nucleic acid capable of inducing RNAi. In further embodiments, the agricultural composition comprises the first minicell encapsulating a nucleic acid that is capable of inducing RNA interference and the second minicell encapsulating the pesticide. In some embodiments, the agricultural composition of the present disclosure further comprises an agricultural suitable additive or adjuvant. [00317] The present disclosure teaches that minicell-encapsulated or minicell-mediated RNA molecule works as biopesticide to reduce a pest (such as insects and fungi) that is resistant to or tolerant of a chemical or biological pesticide. In some embodiments, at minicell- encapsulated or minicell-mediated RNA molecule can kill or control a pest (such as insects and fungi) that is resistant to or tolerant of a chemical or biological pesticide to reduce a number of pesticide-resistant pest population. In some embodiments, at minicell-encapsulated or minicell- mediated RNA molecule can kill or control a pest (such as insects and fungi) have developed resistance to or tolerance of a chemical or biological pesticide to reduce a number of pesticide- resistant pest population. The present disclosure provides minicell-encapsulated or minicell- mediated RNA molecule prevents pests from developing resistance to pesticides. 78 303137700 [00318] The disclosure teaches (i) minicells encapsulated both RNA molecules (such as dsRNA, siRNA, miRNA, and antisense RNA) and chemical pesticides, (ii) minicell encapsulated RNAs paired with unencapsulated chemical pesticides, and (iii) a first minicell encapsulated RNAs paired with a second minicell encapsulated pesticides. [00319] In some embodiments, an agricultural composition comprises a first minicell encapsulating a nucleic acid that is capable of inducing RNA interference in an agricultural pest and a second minicell encapsulating the pesticide that is capable of killing or controlling the agricultural pest, wherein the nucleic acid reduces resistance to or tolerance of a pesticide in the agricultural pest. In some embodiments, an agricultural composition comprises a minicell encapsulating (i) a nucleic acid capable of inducing RNA interference in an agricultural pest and (ii) a pesticide capable of killing or controlling the agricultural pest, wherein the nucleic acid reduces resistance to or tolerance of a pesticide in the agricultural pest. [00320] In some embodiments, an agricultural composition comprises a first minicell encapsulating a single-stranded nucleic acid (e.g., an antisense RNA) and a second minicell encapsulating the pesticide that is capable of killing or controlling the agricultural pest, wherein the single-stranded nucleic acid reduces resistance to or tolerance of a pesticide in an agricultural pest. In some embodiments, agricultural composition, comprising: a minicell encapsulating (i) a single-stranded nucleic acid and (ii) a pesticide capable of killing or controlling an agricultural pest, wherein the single-stranded nucleic acid reduces resistance to or tolerance of a pesticide in the agricultural pest. [00321] The present disclosure provides that this novel approach using a minicell platform for delivering a nucleic acid capable of inducing RNAi to a target along with chemical pesticides. [00322] The present disclosure also provides that this novel approach using a minicell platform for delivering a nucleic acid capable of inducing RNAi to a target along with biopesticides such as protoxins. [00323] Pests can introduce mutations into genes, which confer pesticide resistance to pesticides. In some embodiments, the RNA molecule encapsulated by minicell targets and downregulates expression of genes that are responsible for pesticide resistance. In other embodiments, the RNA molecule encapsulated by minicell targets and downregulates expression of mutated genes conferring pesticide resistance. [00324] In further embodiments, the RNA molecule restores sensitivity or susceptibility to a chemical or biological pesticide in an insect or fungal population by upregulating expression of a target gene via RNA-activation mechanism. 79 303137700 Co-application of RNAi with Pesticides Using Minicell Technology [00325] RNAi is a post-transcription gene regulation mechanism that is present in all known eukaryotes. The cellular RNAi machinery is initiated by dsRNAs that are initially processed into small interfering RNAs (siRNAs) by Dicer-like (DCL) proteins and eventually leads to the degradation of target mRNAs through the action of the gene silencing complex (RISC). RNAi-based genetic transformation technology has widely been utilized to control several insect pests, and diseases, in what is collectively coined as ‘host-induced gene silencing’ (HIGS) (Fire et al 1998; Baulcombe et al 2015). For instance, the expression of dsRNAs targeting dcl1/2 or target of the rapamycin (TOR) genes of B. cinerea significantly suppressed gray mold disease progression in transgenic Arabidopsis, potato and tomato plants, respectively (Xiong et al 2019). [00326] Recently, spray-induced gene silencing (SIGS), which involves the exogenous application of the RNAi has emerged as an appealing alternative to HIGS, as it does not incorporate foreign genes in the treated species (Islam et al 2020; Wang et al 2017; Mcloughlin et al 2018). In some embodiments, the application of dsRNA or siRNA using the minicell platform taught herein is provided to restore one or more pests’ susceptibility to a plurality of pesticides. [00327] Targets of RNAi include but are not limited to genes encoding proteins and/or receptors associated with development of pesticide resistance in a pest or multiple herbicide- resistant weeds/plants. [00328] In some embodiments, dsRNA, antisense RNA, miRNA or siRNA for RNAi are loaded into the minicell platform. The advantage of the minicell platform is that the encapsulation capsule and biomolecule of interest, in this case dsRNA, can both be produced in one fermentation batch. Once the dsRNA is produced and encapsulated in the minicell, the dsRNA is significantly more stable than dsRNA on its own. [00329] In some embodiments, the minicell platform has proven to significantly enhance the stability of dsRNA. The present disclosure also describes a dsRNA bioproduction platform that is based on bacterial minicell carrier systems. [00330] The present disclosure provides the development and applicability of minicell-based RNAi technology in agriculture, in combination with treatment of chemical or biological pesticides. The disclosure presents a robust, scalable platform for producing minicell- encapsulated dsRNAs (ME-dsRNAs) and/or producing minicell-encapsulated pesticides. In some embodiments, ME-dsRNAs are treated with an exogenous pesticide or minicell- encapsulated pesticides. In other embodiments, the present disclosure provides the 80 303137700 development and applicability of minicell encapsulated or loaded with both dsRNAs and pesticides. The minicell platform for delivering dsRNAs and/or pesticides has high stability, efficacy and scalability. [00331] In some embodiments, this minicell platform is incorporated into Integrated Pest Management (IPM) programs which reduce reliance on chemical control and rescue pests’ susceptibility to pesticide resistance. [00332] The present disclosure teaches the use of minicell-based RNAi technology in integrated pest/disease management programs for controlling pests, viruses, and other fungal pathogens and preventing them from development of pesticide resistance, in combination of use of synthetic chemicals or biocontrols. [00333] In some embodiments, Escherichia coli derived anucleated minicells can be utilized as a cost-effective, scalable platform for dsRNA production and encapsulation. [00334] In some embodiments, minicell-encapsulated dsRNA (ME-dsRNA) is shielded from RNase degradation. ME-dsRNAs selectively target genes encoding proteins and/or receptors associated with development of pesticide resistance in a pest or multiple herbicide-resistant weeds/plants, which would lead to the delaying of the development of resistance to transgenic insecticidal crops and/or chemical pesticides, and the rescuing of one or more pests' susceptibility to transgenic insecticidal crops and/or chemical pesticides. [00335] In some embodiments, the potential of ME-dsRNAs to enable the commercial application of RNAi based species-specific biocontrols along with application of conventional chemicals (such as chemical insecticides, fungicides, herbicides, etc.). [00336] In some embodiments, the potential of ME-dsRNAs to enable the commercial application of RNAi based species-specific biocontrols along with application of biological chemicals such as protein toxins. [00337] The present disclosure teaches methods of suppressing development of pesticide resistance in at least one pest or plants of one or more species, the method comprising: applying with at least one pesticide an agricultural formulation comprising a first minicell comprising at least one biologically active compound (e.g., RNA molecule that is a nucleic acid capable of inducing RNAi). [00338] The present disclosure also teaches methods of restoring susceptibility of at least one pest or plants of one or more species to at least one pesticide, the method comprising: the method comprising: applying with at least one pesticide an agricultural formulation comprising a first minicell comprising at least one biologically active compound (e.g., RNA molecule, which is a nucleic acid capable of inducing RNAi or an antisense RNA). 81 303137700 [00339] In some embodiments, the at least one biologically active compound is a nucleic acid which is capable of inducing RNAi. In some embodiments, the at least one biologically active compound is a single-stranded nucleic acid, which is an antisense RNA or an antisense oligonucleotide. In some embodiments, the at least one biologically active compound is a nucleic acid which recognizes a transcript encoding a polypeptide within a cell of a target. In some embodiments, said at least one pesticide is applied exogenously. In other embodiments, said at least one pesticide is loaded into the first minicell comprising the at least one biologically active compound. In further embodiments, said at least one pesticide is loaded into a second minicell and the second minicell is applied to a target pest together with the first minicell comprising a RNA molecule. [00340] In some embodiments, said at least one pesticide is an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide, or an antimicrobial. In other embodiments, said target comprises a plant, an insect, a worm, a bacterium, a fungus, a virus, a nematode, a snail, or a slug. In further embodiments, said nucleic acid is selected from the group consisting of an antisense nucleic acid, a double-stranded RNA (dsRNA), a short-hairpin RNA (shRNA), a small-interfering RNA (siRNA), a microRNA (miRNA), an antisense RNA, a ribozyme, an aptamer, and combination thereof. [00341] The present disclosure teaches application of minicell-encapsulated RNA molecule with a chemical or biological pesticide sequentially or concurrently. The RNA specifically acts on a pesticide-resistant pest to reduce, control, or counteract a built-up resistance, thereby keeping pesticide-resistant pests low in number and maintain a population of pesticide-resistant pests less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, and less than 1% among total pest population composed of pesticide-resistant pests and pesticide-susceptible pests. [00342] In some embodiments, minicell-encapsulated RNA molecule is applied with a chemical or biological insecticide sequentially or concurrently. The RNA specifically acts on an insecticide-resistant pest to reduce, control, or counteract a built-up resistance, thereby keeping insecticide-resistant pests low in number and maintain a population of insecticide- resistant pests less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, and less than 1% 82 303137700 among total pest population composed of insecticide-resistant pests and insecticide-susceptible pests. [00343] In some embodiments, minicell-encapsulated RNA molecule is applied with a chemical or biological fungicide sequentially or concurrently. The RNA specifically acts on an fungicide-resistant pest to reduce, control, or counteract a built-up resistance, thereby keeping fungicide-resistant pests low in number and maintain a population of fungicide-resistant pests less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, and less than 1% among total pest population composed of fungicide-resistant pests and fungicide-susceptible pests. [00344] In some embodiments, minicell-encapsulated RNA molecule is applied with a chemical or biological herbicide sequentially or concurrently. The RNA specifically acts on an herbicide-resistant pest to reduce, control, or counteract a built-up resistance, thereby keeping herbicide-resistant pests low in number and maintain a population of herbicide-resistant pests less than 50%, less than 45%, less than 40%, less than 35%, less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 9%, less than 8%, less than 7%, less than 6%, less than 5%, less than 4%, less than 3%, less than 2%, and less than 1% among total pest population composed of herbicide-resistant pests and herbicide-susceptible pests. Method for suppressing pesticide resistance and restoring susceptibility to pesticide in pests [00345] The present disclosure provides a method of reducing pesticide resistance in an agricultural pest, the method comprising applying an agricultural composition of the present disclosure (such as a minicell encapsulating a nucleic acid capable of inducing RNAi and/or a pesticide, and a minicell encapsulating an antisense RNA and/or a pesticide) to an agricultural pest that are resistant to or tolerant of a pesticide. In some embodiments, the pesticide resistance in the agricultural pest is reduced or suppressed after the application of the agricultural composition. In some embodiments of the methods, pesticide resistance is reduced at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 80%, or at least 100% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. [00346] The present disclosure provides a method of restoring susceptibility of an agricultural pest to a pesticide, the method comprising applying an agricultural composition of the present 83 303137700 disclosure (such as a minicell encapsulating a nucleic acid capable of inducing RNAi and/or a pesticide, and a minicell encapsulating an antisense RNA and/or a pesticide) to an agricultural pest that are resistant to or tolerant of a pesticide, wherein the agricultural pest is restored to be susceptible to a pesticide after the application of the agricultural composition. In some embodiments of the methods, the susceptibility to the pesticide is restored at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 70%, at least 80%, at least 80%, or at least 100% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. In some embodiments of the methods, the agricultural pest applied with the agricultural composition is more sensitive to the pesticide than an agricultural pest unapplied with the agricultural composition. [00347] In some embodiments of the methods, the pesticide is a chemical pesticide or a biological pesticide. In some embodiments of the methods, the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. In some embodiments of the methods, the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. EXAMPLES [00348] The following examples are given for the purpose of illustrating various embodiments of the disclosure and are not meant to limit the present disclosure in any fashion. Changes therein and other uses which are encompassed within the spirit of the disclosure, as defined by the scope of the claims, will occur to those skilled in the art. Example 1. Efficacy of minicell-mediated RNA molecules on Bt-resistant diamondback moth (Plutella xylostella) populations [00349] RNA molecule (i.e., dsRNA) was encapsulated by the minicells of the present disclosure and then applied/treated to Bacillus thuringiensis (Bt)-resistant Diamondback Moth (DBM) populations to assess efficacy of minicell-encapsulated dsRNA on controlling Bt- resistant DBM populations. NO-QAGE strain (obtained from Benzon Research) was used as Bt-resistant DBM population for experiments. This Bt-resistant P. xylostella It was derived from crossing the susceptible "Geneva" strain with a population of field selected P. xylostella 84 303137700 (evolved Bt resistance) originally collected in Hawaii. The strain maintains a level of resistance to various Cry proteins that is approximately 500-1,000 times higher than the susceptible DBM strain. [00350] The second instar (L₂) larvae of Bt-resistant DBM strain (no-QAGE strain) were treated with (1) two separate minicells encapsulating each of target dsRNAs used as bioinsecticides: i) ODBMhB_dsRNA (“ODBMhB”) and ii) ODBMhG_dsRNA (“ODBMhG”); (2) one empty minicell control not encapsulating any dsRNA (“P6NC”); (3) another minicell control encapsulating nontarget-dsRNA (“OBotN”); and (4) Untreated Water Control (“Water”). ODBMhB_dsRNA refers to dsRNA encapsulated by type I of minicell having regular membrane designed for fast release of dsRNA. ODBMhG_dsRNA refers to dsRNA encapsulated by type II of minicell having stronger membrane (treated with glutaraldehyde) designed for controlled, slow release of dsRNA. [00351] 25ng, 50ng, and 100ng of minicells of four test groups (ODBMhB, ODBMhG, P6NC, and OBotN) were treated to ten larvae per treatment (i.e., 10 L₂ larvae). 5 replicates (n = 50 insects per treatment) were run for this bioassay. Three days (72 hours) after treating four test group of minicell-mediated dsRNAs (two target dsRNA, one non-target dsRNA, and no dsRNA) on leaf discs infested with no-QAGE strain, the following analyses were performed to examine absolute mortality, defoliation and insect stunting. Statistical analysis was performed to confirm if the difference observed between the treatment and control are significant. [00352] Figs. 3-4 show percentage of dead DBM larvae in response to feeding on Canola leaf discs coated with ODBMhB_dsRNA and ODBMhG_dsRNA, and two controls (P6NC and OBotN) after 72 hours. Compared to the controls, larvae death rate in response to feeding on minicell-mediated dsRNAs, (1) ODBMhB_dsRNA and (2) ODBMhG_dsRNA was at least 2- 4 folds higher than the controls with empty minicells (P6NC) and minicell-mediated non-target dsRNA (OBotN). Data is presented as ±SEM (n=50) where ***p<0.001; **p<0.01; *p <0.05 and NS =Not significant . Example 2. Efficacy of minicell-mediated RNA molecules on diamide-resistant diamondback moth (Plutella xylostella) populations [00353] RNA molecule (i.e., dsRNA) was encapsulated by the minicells of the present disclosure and then applied/treated to Diamondback Moth (DBM) populations, which are resistant to a chemical pesticide (e.g., diamide-resistant) to assess efficacy of minicell- encapsulated dsRNA on controlling diamide-resistant DBM populations. L1/L2 (36-48 hours 85 303137700 after hatching) of three F1 DBM populations, population 1 – P. xylostella in Takhli; population 2 – P. xylostella in Chiang Mai (CM); population 3 – P. xylostella in Kanchanaburi (KB), were tested for DBM’s susceptibility to chemical insecticides (such as Flubendiamide and Chlorantraniliprole) according to the publicly available Insecticide Resistance Action Committee (IRAC) method No. 18. Table 1 shows diamide resistance of three DBM populations tested in this example. Table 1. Diamide insecticides resistance of DBM populations

[00354] To assess efficacy of minicell-encapsulated dsRNA on controlling diamide-resistant DBM populations, three diamide-resistant DBM populations listed in Table 1 were treated with (1) three separate minicells encapsulating each of target dsRNAs: i) AGS-dsRNA1, ii) AGS- dsRNA2, and iii) AGS-dsRNA3; (2) one empty minicell without dsRNA encapsulated (AGS- NC); (3) untreated control. [00355] Table 2 show numbers of dead DBM larvae in response to feeding on three separate minicells each of which encapsulates dsRNAs (1-3), respectively, in comparison to minicell not encapsulating dsRNAs. Compared to the empty minicell control (AGS-NC), larvae death numbers in response to feeding on minicell-mediated dsRNAs (AGS-dsRNA1, AGS-dsRNA2, and AGS- dsRNA3) were at least 5 folds higher than the minicell control (AGS-NC). Table 2. Efficacy data of dead larvae numbers compiled for RNA (1-3) at 5 Day After Treatment (DAT) using IRAC method 18

Example 3. Selective efficacy of minicell-mediated RNA molecules on Bt-resistant diamondback moth (Plutella xylostella) populations [00356] The experiments similar to Example 1 were conducted to test selective efficacy of the minicell-mediated dsRNAs on Bt-resistant DBM population (Bt.R) and Bt-susceptible DBM 86 303137700 population (Bt.S). Also, slow and fast dsRNA release was tested if there are any impact on efficacy between the slow dsRNA release formulation and the fast dsRNA release formulation. [00357] 100 ng of two empty minicell lines were treated to Bt.R and Bt.S populations as control ; R1G-Control 100ng (slow dsRNA release formulation) and R1B-control 100ng (fast dsRNA release formulation). R1G_Control refers to an empty minicell having regular membrane, but no dsRNA encapsulated as control. R1B_Control refers to an empty minicell having stronger membrane (treated with glutaraldehyde), but no dsRNA encapsulated as control. [00358] 100 ng of four minicell-mediated dsRNAs were treated to Bt.R and Bt.S populations; (i) R1ODBMhG-dsRNA1 (slow dsRNA release formulation) and R1ODBMhB-dsRNA1 (fast dsRNA release formulation); (ii) R1ODBMhNPG-dsRNA2 (slow dsRNA release formulation) and R1ODBMhNPB-dsRNA2 (fast dsRNA release formulation); (iii) R12AG_ dsRNA3 (slow dsRNA release formulation) and R12AB_dsRNA3 (fast dsRNA release formulation); (iv) R12BG_dsRNA4 (slow dsRNA release formulation) and R12BB_dsRNA4 (fast dsRNA release formulation). [00359] Figs.5-6 show percentage of dead DBM larvae in response to feeding on Canola leaf discs coated with 100-125 ng of empty minicell lines and minicell encapsulated dsRNAs after 72 hours (72 hr). Fig.5 shows that dsRNA1 encapsulated by slow release minicell formulation (R1ODBMhG-dsRNA1) led to about 60% DBM larvae death (60% mortality), while the fast release minicell formulation (R1ODBMhB-dsRNA1) had about 20% DBM larvae death (20% mortality) at 72 hours after treatment. Fig.5 shows that dsRNA2 encapsulated by slow release minicell formulation (R1ODBMhNPG-dsRNA2) led to about 68% DBM larvae death (68% mortality), while the fast release minicell formulation (R1ODBMhNPB-dsRNA2) had about 28% DBM larvae death (28% mortality) at 72 hours after treatment. Fig.5 shows that dsRNA3 encapsulated by slow release minicell formulation (R12AG_ dsRNA3) led to about 18% DBM larvae death (18% mortality), while the fast release minicell formulation (R12AB_ dsRNA3) had about 20% DBM larvae death (20% mortality) at 72 hours after treatment. Fig. 5 shows that dsRNA4 encapsulated by slow release minicell formulation (R12BG_dsRNA4) led to about 66% DBM larvae death (66% mortality), while the fast release minicell formulation (R12BB_dsRNA4) had about 62% DBM larvae death (62% mortality) at 72 hours after treatment. [00360] Data is presented as ±SEM (n=30 (Bt.S) and n=50 (Bt.R) and the % differences in death between the controls and treatments at 50% threshold are significant between ***p<0.001 & **p<0.01. The response of Bt-resistant (left bars) and Bt-susceptible (right bars) 87 303137700 DBM larvae is presented in Fig. 5. Also, slow dsRNA release formulation and fast dsRNA release formulation are represented by solid and shaded bars in Fig. 5, respectively. Example 4. Mechanism of action of minicell-mediated RNA molecules qPCR Study: Studying transcript degradation & correlating to insecticidal phenotype [00361] To understand target transcriptional changes involved with insecticide resistance developed in pests mechanisms when minicell-mediated dsRNA is treated to the insecticide- resistant insects, larvae of Plutella xylostella were treated with 12.5 ng and 100 ng of minicell- mediated dsRNA (ODBMhG_dsRNA slow release formulation) over 24 and 72 hours. Also, 12.5 ng and 100 ng of one empty minicell encapsulating no dsRNA (R1NC) was treated to larvae of Plutella xylostella. [00362] When 100 ng of ODBMhG_dsRNA was fed to larvae of insecticide-resistant Plutella xylostella, about 76% of the larvae were dead in comparison of about 38% death of larvae fed with 100 ng of R1NC. From the dead larvae fed with minicell formulations (R1NC and ODBMhG_dsRNA), the mRNA was extracted and followed by RT-qPCT to examine expression level of two genes related to pesticide resistance; (1) Ryanodine receptor gene that is the target binding site for diamide chemicals; (2) UDP-glucuronosyltransferase receptor gene that plays a role in detoxifying chemical insecticides. [00363] The RT-qPCR data as presented in Figs. 7A and 7B are from three independent experiments 1, 2 and 3, (3 replicates; n=10). Fig. 7A shows relative fold change of the target transcript (i.e., ryanodine receptor 44F-like gene transcript) downregulation in comparison to EF1A internal control transcript in the larvae feeding ODBMhG_dsRNAs and controls at various time points. Data is presented with transformed ±SEM of 3 replicates (n=10). The fold change differences of the target transcript downregulation between R1NC treatment and ODBMhG treatment are at least 2 folds, which are significant at **p<0.01 and *p <0.05. Also, the difference of larvae death percentage(%) between 100ng of empty minicell control (R1NC) treatment (38% death) and 100ng of minicell-mediated target dsRNA (ODBMhG_dsRNA) treatment (76% death) are at least 2 folds, which correspond to the fold changes of the target transcript. [00364] Fig. 7B shows relative fold change of the target transcript (i.e., UDP- glucuronosyltransferase receptor gene transcript) downregulation in comparison to EF1A internal control transcript in the larvae feeding ODBMhG_dsRNAs and controls at various time points. Data is presented with transformed ±SEM of 3 replicates (n=10). The fold change differences of the target transcript downregulation between R1NC treatment and ODBMhG 88 303137700 treatment are at least 1.5 folds, which are significant at **p<0.01 and *p <0.05. Also, the difference of larvae death percentage(%) between 100ng of empty minicell control (R1NC) treatment (38% death) and 100ng of minicell-mediated target dsRNA (ODBMhG) treatment (76% death) are at least 2 folds, which are comparable to the fold changes of the target transcript. [00365] Observed transcript downregulation in Figs. 7A and 7B corelates to increased insecticidal phenotype of larvae death in the insects responding to ODBMhG dsRNA. Figs.7A and 7B indicate that 100 ng of ODBMhG dsRNA caused about 4-8 fold down regulation of target transcripts by 72-hour post ingestion. Foliar applied minicell-mediated dsRNA (ODBMhG_dsRNA) downregulates target transcripts to confer insecticidal activity. [00366] Both ryanodine receptor 44F-like gene and UDP-glucuronosyltransferase receptor gene play key roles in developing pesticide/insecticide resistance in pests. [00367] Ryanodine receptor is the target binding site for diamide chemicals. Overexpression of Ryanodine receptor or introduced mutation in the P. xylostella ryanodine receptor (e.g., G4946E) are linked to the high level of resistance to diamides in populations. First, targeting DBM populations with polymorphisms that confer resistance is a mechanism of prevent those genotypes from proliferating and maintain a higher level of susceptible genotypes in the population. Second, downregulating Ryanodine receptor expression is another mechanism of counteracting high levels of Ryanodine receptor expression. [00368] UDP-glucuronosyltransferase receptor has a role in directly detoxifying chemical insecticides. By downregulating expression of UDP-glucuronosyltransferase receptor gene and similar detoxification genes (such as cytochrome P450 monooxygenase (P450), carboxylesterase (CarE) and glutathione S-transferase (GSTs) , pests can lose their sensitivity to pesticides including insecticides, Thus, sensitivity is enhanced in pest populations by downregulating or repressing these genes such as ryanodine receptor 44F-like gene and UDP- glucuronosyltransferase receptor gene. Example 5. Lab studies for synergistic effect of co-application of minicell-mediated dsRNA and chemical pesticides [00369] This example describes the enhanced efficacy of co-application minicell-mediated dsRNA and chemical insecticide (Coragen®) at low doses. [00370] 2.5 cm diameter cabbage leaf disc with 4 replications (4 leaf discs per treatment) were prepared. At Day 1, cabbage leaf discs were treated with 200ul of formulation product. Each leaf disc was treated with 100ul on the top of the leaf, and allowed to dry at room temperature. 89 303137700 Also, 100ul was treated on the bottom of the leaf, and allowed to air dry at room temperature. ach leaf disc was placed in individual Petri dishes lined with filter paper. Ten DBM (1^st instar larvae acquired from Benzon Research) were placed on each leaf disc. [00371] At Day 2, mortality counts (numbers of dead DBM) were made at 24 hours after the treatment. Larvae considered dead if discolored or if does not respond to gentle prodding with a fine brush. New leaf discs were treated in a condition same as for Day 1. Live larvae were transferred from day 1 leaf discs to day 2 leaf discs. [00372] At Day 3, methods from day 2 were repeated at 48 hours after the initial treatment. Final mortality counts made 24 hours after larvae are transferred to day 3 leaf discs (i.e., 72 hour). Table 3 shows mean number of dead DBM larvae per treatment at 24, 48, and 72 hour time points. Minicell-dsRNA-B refers to dsRNA molecule encapsulated by minicell having regular membrane designed for fast release. Minicell-dsRNA-G refers to refers to dsRNA molecule encapsulated by minicell having glutaraldehyde-treated membrane designed for slow release. Naked RNA refers to RNA molecule not protected and/or encapsulated by minicell. [00373] Treatment No.10 in Table 3 presents that 5 ng of minicell-RNA + Coragen® 12.5ul/l were co-applied twice at 0 and 24 hours, and then DBM mortality was measured at 48 hours from the initial treatment. This treatment No.10 demonstrates the highest dead rate (7.8 out of 10 DBMs) among other treatments, indicating synergistic effect of co-application of minicell- mediated RNA and chemical insecticide at low dose on controlling insects (e.g., DBM). [00374] Also, Treatment No. 15 in Table 3 presents that 5 ng of minicell-RNA + Coragen® 3ul/l were co-applied twice at 0 and 24 hours, and then DBM mortality was measured at 48 hours from the initial treatment. When comparing treatment No. 15 to treatment No. 16 (only Coragen® 3.0ul/l), this co-application in treatment No. 15 demonstrates superior effects (at least 2 fold higher than) over treatment No. 16. Even the DBM mortality of treatment No. 15 is comparable to a very high dose of Coragen® (25.0ul/l). This data also indicates synergistic effect of co-application of minicell-mediated RNA and chemical insecticide at low dose on controlling insects (e.g., DBM). 0.25% Dyne-Amic surfactant was added to each treatment. 90 303137700 Table 3. Mean number of dead DBM larvae at each 24-hour interval.

Example 6. Natural infestation field studies for synergistic effect of co-application of minicell-mediated dsRNA and chemical pesticides [00375] While the experiments described in Example 5 were performed in a lab setting, similar experiments were conducted in a field. Four-week old transplants were prepared. Individual plots are 20 feet long on single-row plastic beds with 15-inch in row spacing and trickle irrigation. Cabbage cultivar used for the field experiments was 'Blue Dynasty'. Each treatment was replicated four times in a randomized block design. Insecticide treatment was made Day 1 (1^st day of week 1), Day 8 (1^st day of week 2), Day 15 (1^st day of week 3), and Day 22 (1^st day of week 4) at 1-week interval. Treatments applied with a CO₂ sprayer using three hollow cone 91 303137700 nozzles (overhead and on each side of the plant at 45 psi which deliver 1200 mls per treatment). Data on percent plant defoliation and insect counts were taken from five plants per plot on each sampling date. 0.25% Dyne-Amic surfactant was added to each treatment. Minicell-dsRNA- HP refers to dsRNA-HP (dsRNA hairpin) encapsulated by minicell of the present disclosure. [00376] Tables 4-8 present percentage of defoliation at Weeks 1-5 on 3 and 6 day after treatment (DAT) based on combinations of applications using minicell-dsRNA along with chemical insecticide (Coragen®) and or biological insecticide (Xentari®). Table 4. Percentage of defoliation at Week 1 on 3 and 6 day after treatment (DAT)

Table 5. Percentage of defoliation at Week 2 on 3 and 6 day after treatment (DAT)

Table 6. Percentage of defoliation at Week 3 on 3 and 6 day after treatment (DAT)

92 303137700

Table 7. Percentage of defoliation at Week 4 on 3 and 6 day after treatment (DAT)

Table 8. Percentage of defoliation at Week 5 on 3 and 6 day after treatment (DAT)

[00377] Figs. 8A-8H show cabbage defoliation at eight treatments as presented in Table 8; (1) Untreated Check (Fig.8A) showing about 11% defoliation by week 5, (2) Coragen® + Dyne- Amic (Fig. 8B) showing no sign of defoliation by week 5, (3) Xentari® + Dyne-Amic (Fig. 8C) showing <1% defoliation by week 5, (4) Minicell-dsRNA HP-High + Dyne-Amic (Fig. 8D) showing about 7% defoliation by week 5, (5) Minicell-dsRNA HP-Low + Dyne-Amic (Fig. 8E) showing about 4-5% defoliation by week 5, (6) Coragen® + Dyne-Amic (Week 1) & Minicell- 93 303137700 dsRNA HP-Low (Week 2) (Fig. 8F) showing <1% defoliation by week 5, (7) Minicell-dsRNA HP-Low + Xentari® + Dyne-Amic (Fig.8G) showing <1% defoliation by week 5, (8) Xentari® + Dyne-Amic (Week 1) & Minicell-dsRNA HP-Low (Week 2) (Fig. 8H) showing about 1-2% defoliation by week 5. [00378] Minicell-dsRNA treatment in integrated pest management (IPM) rotations can reduce use of chemical insecticide (e.g., Coragen®, Xentari®) and/or biological insecticide (e.g., Bt toxin, Cry toxin) applied by 75%, while maintaining <1% defoliation. [00379] Defoliation data from diamondback pressure was interpreted carefully due to confounding effects of concurrent fall armyworm, yellowstriped armyworm and imported cabbageworm infestations. 1) Rotational Xentari (1/4 rate) with Minicell-dsRNA-HP low performed better than the full rate of Xentari and statistically comparable to Coragen 8 fl/oz acre. This suggests the Minicell-dsRNA-HP formulation/product has synergy with Xentari and can perform similar to the synthetic standard.2) Tank mix with Xentari (1/2 rate) with Minicell- dsRNA-HP performs similar to full rate of Coragen 8 fl/oz acre. This suggests that the Minicell- dsRNA formulation/product can be used as a tank mix in conjunction with Xentari. 3) Rotational with Coragen (4 fl/oz) and Minicell-dsRNA-HP formulation/product performs similar to full rate of Coragen 8 fl/oz acre. This suggests that the Minicell-dsRNA-HP formulation/product can be used in conjunction with Coragen to reduce the development of resistance with this valuable synthetic chemical. Most importantly, this aligns with reducing synthetic pesticide applications. 4) Average defoliation in plots is less than 5%. Example 7. Efficacy of minicell-mediated RNA molecules on succinate dehydrogenase inhibitors (SDHI) fungicide-resistant fungi models [00380] 1, 10, and 50 ug/ml of minicell-mediated dsRNA molecules (R1, R2, and R3) were treated to highly SDHI fungicide-resistant fungi (FL12-355, 82-10, GP18-205) on controlling SDHI-resistant fungi populations in comparison to an empty minicell (M). Also, wild type fungi (4E1 and VT were treated with 1, 10, and 50 ug/ml of R1, R2, R3, and M. [00381] Fig. 9 shows in-vitro mycelial growth inhibition by minicell-RNA against highly resistant to SDHI strains of B. cinerea, indicating that minicell-mediated RNA molecule can be used to inhibit fungi having fungicide resistance. [00382] While various inventive embodiments have been described and illustrated herein, those of ordinary skill in the art will readily envision a variety of other means and/or structures for performing the function and/or obtaining the results and/or one or more of the advantages described herein, and each of such variations and/or modifications is deemed to be within the 94 303137700 scope of the inventive embodiments described herein. More generally, those skilled in the art will readily appreciate that all parameters, dimensions, materials, and configurations described herein are meant to be exemplary and that the actual parameters, dimensions, materials, and/or configurations will depend upon the specific application or applications for which the inventive teachings is/are used. Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific inventive embodiments described herein. It is, therefore, to be understood that the foregoing embodiments are presented by way of example only and that, within the scope of the appended claims and equivalents thereto; inventive embodiments may be practiced otherwise than as specifically described and claimed. Inventive embodiments of the present disclosure are directed to each individual feature, system, article, material, kit, and/or method described herein. In addition, any combination of two or more such features, systems, articles, materials, kits, and/or methods, if such features, systems, articles, materials, kits, and/or methods are not mutually inconsistent, is included within the inventive scope of the present disclosure. [00383] Also, various inventive concepts may be embodied as one or more methods, of which examples have been provided. The acts performed as part of the methods may be ordered in any suitable way. Accordingly, embodiments may be constructed in which acts are performed in an order different than illustrated, which may include performing some acts simultaneously, even though shown as sequential acts in illustrative embodiments. NUMBERED EMBODIMENTS OF THE DISCLOSURE [00384] Notwithstanding the appended claims, the disclosure sets forth the following numbered embodiments: Nucleic acid that is capable of inducing RNA interference 1. An agricultural composition, comprising: a. a first minicell encapsulating a nucleic acid that is capable of inducing RNA interference in an agricultural pest, wherein the nucleic acid reduces resistance to or tolerance of a pesticide in the agricultural pest. 2. The agricultural composition of embodiment 1, wherein the agricultural pest is resistant to or tolerant of the pesticide. 95 303137700 3. The agricultural composition of embodiment 1 or 2, wherein the pesticide is a chemical pesticide or a biological pesticide. 4. The agricultural composition of any one of embodiments 1-3, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. 5. The agricultural composition of embodiment 1 or 2, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. 6. The agricultural composition of embodiment 1, wherein the nucleic acid is capable of recovering the agricultural pest’s sensitivity or susceptibility to the pesticide. 7. The agricultural composition of embodiment 1, wherein the nucleic acid is capable of altering expression of a gene responsible for pesticide resistance or tolerance. 8. The agricultural composition of embodiment 7, wherein the expression of the gene responsible for pesticide resistance or tolerance is downregulated. 9. The agricultural composition of embodiment 7, wherein the expression of the gene responsible for pesticide resistance or tolerance is upregulated. 10. The agricultural composition of embodiment 7, wherein the gene responsible for pesticide resistance or tolerance is an ion channel gene, a detoxification gene, a target site resistance gene, or a transporter gene. 11. The agricultural composition of embodiment 10, wherein the ion channel gene is a gene encoding Ryanodine receptor (RyR) or Voltage-gated sodium channel (VGSC). 12. The agricultural composition of embodiment 10, wherein the detoxification gene is selected from the group consisting of a gene encoding UDP-glycosyltransferase (UGT), Cytochrome P450 monooxygenase, Esterase, Carboxylesterase (CarE), and Glutathione S- transferase (GST). 13. The agricultural composition of embodiment 10, wherein the target site resistance gene is selected from the group consisting of a gene encoding Acetylcholinesterase (AChE), Voltage-gated sodium channel (VGSC), Gamma-aminobutyric acid (GABA) receptor, Nicotinic acetylcholine receptor (nAChR), and Glutamate-gated chloride channel (GluCl). 14. The agricultural composition of embodiment 10, wherein the transporter gene is selected from the group consisting of a gene encoding ATP-binding cassette (ABC) transporter, Solute carrier (SLC) transporter, Major facilitator superfamily (MFS) transporter, and P- glycoprotein. 96 303137700 15. The agricultural composition of any one of embodiments 1-14, wherein the nucleic acid is capable of inducing RNA interference in at least one member from an order selected from the group consisting of: Coleoptera, Diptera, Hymenoptera, Lepidoptera, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Siphonaptera, and Trichoptera. 16. The agricultural composition of any one of embodiments 1-15, wherein the nucleic acid is capable of inducing RNA interference in a member of the order Lepidoptera. 17. The agricultural composition of any one of embodiments 1-15, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Plutella. 18. The agricultural composition of any one of embodiments 1-15, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Spodoptera. 19. The agricultural composition of any one of embodiments 1-18, wherein the nucleic acid is a RNA molecule. 20. The agricultural composition of any one of embodiments 1-19, wherein the nucleic acid is at least one selected from the group consisting of: a double-stranded RNA (dsRNA) a short- hairpin RNA (shRNA), a small-interfering RNA (siRNA), and a microRNA (miRNA). 21. The agricultural composition of any one of embodiments 1-20, wherein the nucleic acid is dsRNA. 22. The agricultural composition of any one of embodiments 1-20, wherein the nucleic acid is shRNA. 23. The agricultural composition of any one of embodiments 1-20, wherein the nucleic acid is siRNA. 24. The agricultural composition of any one of embodiments 1-20, wherein the nucleic acid is miRNA. 25. The agricultural composition of any one of embodiments 1-24, wherein the minicell is ribonuclease deficient. 26. The agricultural composition of any one of embodiments 1-25, wherein the minicell comprises at least one fusion protein. 27. The agricultural composition of any one of embodiments 1-26, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell. 28. The agricultural composition of any one of embodiments 1-27, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising at least one target cell adhesion moiety. 97 303137700 29. The agricultural composition of any one of embodiments 1-28, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising a carbohydrate binding molecule. 30. The agricultural composition of embodiment 3, wherein the biological pesticide is a protein toxin. 31. The agricultural composition of embodiment 1, further comprising: b. a second minicell encapsulating the pesticide that is capable of killing or controlling the agricultural pest. 32. The agricultural composition of any one of embodiments 1-31, further comprising a solid, dry, or liquid carrier. 33. The agricultural composition of embodiment 32, wherein said solid carrier is in a form of granule or pellet and is selected from the group consisting of: diatomaceous earth, loam, silica, alginate, clay, bentonite, vermiculite, and combinations thereof. 34. The agricultural composition of embodiment 32, wherein said dry carrier in a form of powder and is selected from the group consisting of: peat, wheat, bran, vermiculite, clay mineral, calcium carbonate, dolomite, gypsum, bentonite, rock phosphate, phosphorous compound, titanium dioxide, humus, talc, alginate, activated charcoal, and combinations thereof. 35. The agricultural composition of embodiment 32, wherein said liquid carrier is in a form of liquid or emulsion, and is selected from the group consisting of a surfactant, an emulsifier, a crop oil concentrate, a penetrant, and combinations thereof. 36. An agricultural composition, comprising: a minicell encapsulating (i) a nucleic acid capable of inducing RNA interference in an agricultural pest and (ii) a pesticide capable of killing or controlling the agricultural pest, wherein the nucleic acid reduces resistance to or tolerance of a pesticide in the agricultural pest. 37. The agricultural composition of embodiment 36, wherein the agricultural pest is resistant to or tolerant of the pesticide. 38. The agricultural composition of embodiment 36, wherein the pesticide is a chemical pesticide or a biological pesticide. 39. The agricultural composition of any one of embodiments 36-38, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. 98 303137700 40. The agricultural composition of any one of embodiments 36-39, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. 41. The agricultural composition of embodiment 36, wherein the nucleic acid is capable of recovering the agricultural pest’s sensitivity or susceptibility to the pesticide. 42. The agricultural composition of embodiment 36, wherein the nucleic acid is capable of altering expression of a gene responsible for pesticide resistance or tolerance. 43. The agricultural composition of embodiment 42, wherein the expression of the gene responsible for pesticide resistance or tolerance is downregulated. 44. The agricultural composition of embodiment 42, wherein the expression of the gene responsible for pesticide resistance or tolerance is upregulated. 45. The agricultural composition of embodiment 42, wherein the gene responsible for pesticide resistance or tolerance is an ion channel gene, a detoxification gene, a target site resistance gene, or a transporter gene. 46. The agricultural composition of embodiment 45, wherein the ion channel gene is a gene encoding Ryanodine receptor (RyR) or Voltage-gated sodium channel (VGSC). 47. The agricultural composition of embodiment 45, wherein the detoxification gene is selected from the group consisting of a gene encoding UDP-glycosyltransferase (UGT), Cytochrome P450 monooxygenase, Esterase, Carboxylesterase (CarE), and Glutathione S- transferase (GST). 48. The agricultural composition of embodiment 45, wherein the target site resistance gene is selected from the group consisting of a gene encoding Acetylcholinesterase (AChE), Voltage-gated sodium channel (VGSC), Gamma-aminobutyric acid (GABA) receptor, Nicotinic acetylcholine receptor (nAChR), and Glutamate-gated chloride channel (GluCl). 49. The agricultural composition of embodiment 45, wherein the transporter gene is selected from the group consisting of a gene encoding ATP-binding cassette (ABC) transporter, Solute carrier (SLC) transporter, Major facilitator superfamily (MFS) transporter, and P- glycoprotein. 50. The agricultural composition of any one of embodiments 36-49, wherein the nucleic acid is capable of inducing RNA interference in at least one member from an order selected from the group consisting of: Coleoptera, Diptera, Hymenoptera, Lepidoptera, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Siphonaptera, and Trichoptera. 51. The agricultural composition of any one of embodiments 36-50, wherein the nucleic acid is capable of inducing RNA interference in a member of the order Lepidoptera. 99 303137700 52. The agricultural composition of any one of embodiments 36-50, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Plutella. 53. The agricultural composition of any one of embodiments 36-50, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Spodoptera. 54. The agricultural composition of any one of embodiments 36-53, wherein the nucleic acid is a RNA molecule. 55. The agricultural composition of any one of embodiments 36-54, wherein the nucleic acid is at least one selected from the group consisting of: a double-stranded RNA (dsRNA), a short-hairpin RNA (shRNA), a small-interfering RNA (siRNA), and a microRNA (miRNA). 56. The agricultural composition of any one of embodiments 36-55, wherein the nucleic acid is dsRNA. 57. The agricultural composition of any one of embodiments 36-55, wherein the nucleic acid is shRNA. 58. The agricultural composition of any one of embodiments 36-55, wherein the nucleic acid is siRNA. 59. The agricultural composition of any one of embodiments 36-55, wherein the nucleic acid is miRNA. 60. The agricultural composition of any one of embodiments 36-59, wherein the minicell is ribonuclease deficient. 61. The agricultural composition of any one of embodiments 36-60, wherein the minicell comprises at least one fusion protein. 62. The agricultural composition of any one of embodiments 36-61, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell. 63. The agricultural composition of any one of embodiments 36-62, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising at least one target cell adhesion moiety. 64. The agricultural composition of any one of embodiments 36-63, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising a carbohydrate binding molecule. 65. The agricultural composition of embodiment 38, wherein the biological pesticide is a protein toxin. 66. The agricultural composition of any one of embodiments 36-65, further comprising a solid, dry, or liquid carrier. 100 303137700 67. The agricultural composition of embodiment 66, wherein said solid carrier is in a form of granule or pellet and is selected from the group consisting of: diatomaceous earth, loam, silica, alginate, clay, bentonite, vermiculite, and combinations thereof. 68. The agricultural composition of embodiment 66, wherein said dry carrier in a form of powder and is selected from the group consisting of: peat, wheat, bran, vermiculite, clay mineral, calcium carbonate, dolomite, gypsum, bentonite, rock phosphate, phosphorous compound, titanium dioxide, humus, talc, alginate, activated charcoal, and combinations thereof. 69. The agricultural composition of embodiment 66, wherein said liquid carrier is in a form of liquid or emulsion, and is selected from the group consisting of a surfactant, an emulsifier, a crop oil concentrate, a penetrant, and combinations thereof. 70. A method of reducing or suppressing pesticide resistance in an agricultural pest, the method comprising: applying an agricultural composition of embodiment 1 or 36 to an agricultural pest, wherein resistance to a pesticide in the agricultural pest is reduced or suppressed after the application of the agricultural composition. 71. The method of embodiment 70, wherein the agricultural pest is resistant to or tolerant of the pesticide. 72. The method of embodiment 70, wherein the resistance to the pesticide is reduced at least 10% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. 73. The method of embodiment 70, wherein the pesticide is a chemical pesticide or a biological pesticide. 74. The method of embodiment 73, wherein the biological pesticide is a protein toxin. 75. The method of embodiment 70, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. 76. The method of embodiment 70, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. 77. A method of restoring susceptibility of an agricultural pest to a pesticide, the method comprising: applying an agricultural composition of embodiment 1 or 36 to an agricultural pest, wherein the agricultural pest is restored to be susceptible to a pesticide after the application of the agricultural composition. 101 303137700 78. The method of embodiment 77, wherein the agricultural pest is resistant to or tolerant of the pesticide. 79. The method of embodiment 77, wherein the susceptibility to the pesticide is restored at least 10% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. 80. The method of embodiment 77, wherein the agricultural pest applied with the agricultural composition is more sensitive to the pesticide than an agricultural pest unapplied with the agricultural composition. 81. The method of embodiment 77, wherein the pesticide is a chemical pesticide or a biological pesticide. 82. The method of embodiment 81, wherein the biological pesticide is a protein toxin. 83. The method of embodiment 77, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. 84. The method of embodiment 77, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. Single-stranded antisense oligonucleotide 1. An agricultural composition, comprising: a. a first minicell encapsulating a single-stranded nucleic acid, wherein the nucleic acid reduces resistance to or tolerance of a pesticide in an agricultural pest. 2. The agricultural composition of embodiment 1, wherein the agricultural pest is resistant to or tolerant of the pesticide. 3. The agricultural composition of embodiment 1 or 2, wherein the pesticide is a chemical pesticide or a biological pesticide. 4. The agricultural composition of any one of embodiments 1-3, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. 102 303137700 5. The agricultural composition of embodiment 1 or 2, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. 6. The agricultural composition of embodiment 1, wherein the nucleic acid is capable of recovering the agricultural pest’s sensitivity or susceptibility to the pesticide. 7. The agricultural composition of embodiment 1, wherein the nucleic acid is capable of altering expression of a gene responsible for pesticide resistance or tolerance. 8. The agricultural composition of embodiment 7, wherein the expression of the gene responsible for pesticide resistance or tolerance is downregulated. 9. The agricultural composition of embodiment 7, wherein the gene responsible for pesticide resistance or tolerance is an ion channel gene, a detoxification gene, a target site resistance gene, or a transporter gene. 10. The agricultural composition of embodiment 9, wherein the ion channel gene is a gene encoding Ryanodine receptor (RyR) or Voltage-gated sodium channel (VGSC). 11. The agricultural composition of embodiment 9, wherein the detoxification gene is selected from the group consisting of a gene encoding UDP-glycosyltransferase (UGT), Cytochrome P450 monooxygenase, Esterase, Carboxylesterase (CarE), and Glutathione S- transferase (GST). 12. The agricultural composition of embodiment 9, wherein the target site resistance gene is selected from the group consisting of a gene encoding Acetylcholinesterase (AChE), Voltage-gated sodium channel (VGSC), Gamma-aminobutyric acid (GABA) receptor, Nicotinic acetylcholine receptor (nAChR), and Glutamate-gated chloride channel (GluCl). 13. The agricultural composition of embodiment 9, wherein the transporter gene is selected from the group consisting of a gene encoding ATP-binding cassette (ABC) transporter, Solute carrier (SLC) transporter, Major facilitator superfamily (MFS) transporter, and P- glycoprotein. 14. The agricultural composition of any one of embodiments 1-13, wherein the nucleic acid is capable of inducing RNA interference in at least one member from an order selected from the group consisting of: Coleoptera, Diptera, Hymenoptera, Lepidoptera, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Siphonaptera, and Trichoptera. 15. The agricultural composition of any one of embodiments 1-14, wherein the nucleic acid is capable of inducing RNA interference in a member of the order Lepidoptera. 16. The agricultural composition of any one of embodiments 1-14, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Plutella. 103 303137700 17. The agricultural composition of any one of embodiments 1-14, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Spodoptera. 18. The agricultural composition of any one of embodiments 1-17, wherein the nucleic acid is a RNA molecule. 19. The agricultural composition of any one of embodiments 1-18, wherein the nucleic acid is an antisense RNA. 20. The agricultural composition of any one of embodiments 1-19, wherein the minicell is ribonuclease deficient. 21. The agricultural composition of any one of embodiments 1-20, wherein the minicell comprises at least one fusion protein. 22. The agricultural composition of any one of embodiments 1-21, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell. 23. The agricultural composition of any one of embodiments 1-22, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising at least one target cell adhesion moiety. 24. The agricultural composition of any one of embodiments 1-23, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising a carbohydrate binding molecule. 25. The agricultural composition of embodiment 3, wherein the biological pesticide is a protein toxin. 26. The agricultural composition of embodiment 1, further comprising: b. a second minicell encapsulating the pesticide that is capable of killing or controlling the agricultural pest. 27. The agricultural composition of any one of embodiments 1-26, further comprising a solid, dry, or liquid carrier. 28. The agricultural composition of embodiment 27, wherein said solid carrier is in a form of granule or pellet and is selected from the group consisting of: diatomaceous earth, loam, silica, alginate, clay, bentonite, vermiculite, and combinations thereof. 29. The agricultural composition of embodiment 28, wherein said dry carrier in a form of powder and is selected from the group consisting of: peat, wheat, bran, vermiculite, clay mineral, calcium carbonate, dolomite, gypsum, bentonite, rock phosphate, phosphorous compound, titanium dioxide, humus, talc, alginate, activated charcoal, and combinations thereof. 104 303137700 30. The agricultural composition of embodiment 29, wherein said liquid carrier is in a form of liquid or emulsion, and is selected from the group consisting of a surfactant, an emulsifier, a crop oil concentrate, a penetrant, and combinations thereof. 31. An agricultural composition, comprising: a minicell encapsulating (i) a single-stranded nucleic acid and (ii) a pesticide capable of killing or controlling an agricultural pest, wherein the single-stranded nucleic acid reduces resistance to or tolerance of a pesticide in the agricultural pest. 32. The agricultural composition of embodiment 31, wherein the agricultural pest is resistant to or tolerant of the pesticide. 33. The agricultural composition of embodiment 31, wherein the pesticide is a chemical pesticide or a biological pesticide. 34. The agricultural composition of any one of embodiments 31-33, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. 35. The agricultural composition of any one of embodiments 31-34, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. 36. The agricultural composition of embodiment 31, wherein the nucleic acid is capable of recovering the agricultural pest’s sensitivity or susceptibility to the pesticide. 37. The agricultural composition of embodiment 31, wherein the nucleic acid is capable of altering expression of a gene responsible for pesticide resistance or tolerance. 38. The agricultural composition of embodiment 37, wherein the expression of the gene responsible for pesticide resistance or tolerance is downregulated. 39. The agricultural composition of embodiment 38, wherein the gene responsible for pesticide resistance or tolerance is an ion channel gene, a detoxification gene, a target site resistance gene, or a transporter gene. 40. The agricultural composition of embodiment 39, wherein the ion channel gene is a gene encoding Ryanodine receptor (RyR) or Voltage-gated sodium channel (VGSC). 41. The agricultural composition of embodiment 39, wherein the detoxification gene is selected from the group consisting of a gene encoding UDP-glycosyltransferase (UGT), Cytochrome P450 monooxygenase, Esterase, Carboxylesterase (CarE), and Glutathione S- transferase (GST). 105 303137700 42. The agricultural composition of embodiment 39, wherein the target site resistance gene is selected from the group consisting of a gene encoding Acetylcholinesterase (AChE), Voltage-gated sodium channel (VGSC), Gamma-aminobutyric acid (GABA) receptor, Nicotinic acetylcholine receptor (nAChR), and Glutamate-gated chloride channel (GluCl). 43. The agricultural composition of embodiment 39, wherein the transporter gene is selected from the group consisting of a gene encoding ATP-binding cassette (ABC) transporter, Solute carrier (SLC) transporter, Major facilitator superfamily (MFS) transporter, and P- glycoprotein. 44. The agricultural composition of any one of embodiments 31-43, wherein the nucleic acid is capable of inducing RNA interference in at least one member from an order selected from the group consisting of: Coleoptera, Diptera, Hymenoptera, Lepidoptera, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Siphonaptera, and Trichoptera. 45. The agricultural composition of any one of embodiments 31-44, wherein the nucleic acid is capable of inducing RNA interference in a member of the order Lepidoptera. 46. The agricultural composition of any one of embodiments 31-44, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Plutella. 47. The agricultural composition of any one of embodiments 31-44, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Spodoptera. 48. The agricultural composition of any one of embodiments 31-47, wherein the nucleic acid is a RNA molecule. 49. The agricultural composition of any one of embodiments 31-48, wherein the nucleic acid is an antisense RNA. 50. The agricultural composition of any one of embodiments 31-49, wherein the minicell is ribonuclease deficient. 51. The agricultural composition of any one of embodiments 31-50, wherein the minicell comprises at least one fusion protein. 52. The agricultural composition of any one of embodiments 31-51, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell. 53. The agricultural composition of any one of embodiments 31-52, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising at least one target cell adhesion moiety. 54. The agricultural composition of any one of embodiments 31-53, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising a carbohydrate binding molecule. 106 303137700 55. The agricultural composition of embodiment 33, wherein the biological pesticide is a protein toxin. 56. The agricultural composition of any one of embodiments 31-55, further comprising a solid, dry, or liquid carrier. 57. The agricultural composition of embodiment 56, wherein said solid carrier is in a form of granule or pellet and is selected from the group consisting of: diatomaceous earth, loam, silica, alginate, clay, bentonite, vermiculite, and combinations thereof. 58. The agricultural composition of embodiment 56, wherein said dry carrier in a form of powder and is selected from the group consisting of: peat, wheat, bran, vermiculite, clay mineral, calcium carbonate, dolomite, gypsum, bentonite, rock phosphate, phosphorous compound, titanium dioxide, humus, talc, alginate, activated charcoal, and combinations thereof. 59. The agricultural composition of embodiment 56, wherein said liquid carrier is in a form of liquid or emulsion, and is selected from the group consisting of a surfactant, an emulsifier, a crop oil concentrate, a penetrant, and combinations thereof. 60. A method of reducing or suppressing pesticide resistance in an agricultural pest, the method comprising: applying an agricultural composition of embodiment 1 or 31 to an agricultural pest, wherein resistance to a pesticide in the agricultural pest is reduced or suppressed after the application of the agricultural composition. 61. The method of embodiment 60, wherein the agricultural pest is resistant to or tolerant of the pesticide. 62. The method of embodiment 60, wherein the resistance to the pesticide is reduced at least 10% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. 63. The method of embodiment 60, wherein the pesticide is a chemical pesticide or a biological pesticide. 64. The method of embodiment 63, wherein the biological pesticide is a protein toxin. 65. The method of embodiment 60, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. 66. The method of embodiment 60, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. 107 303137700 67. A method of restoring susceptibility of an agricultural pest to a pesticide, the method comprising: applying an agricultural composition of embodiment 1 or 31 to an agricultural pest, wherein the agricultural pest is restored to be susceptible to a pesticide after the application of the agricultural composition. 68. The method of embodiment 67, wherein the agricultural pest is resistant to or tolerant of the pesticide. 69. The method of embodiment 67, wherein the susceptibility to the pesticide is restored at least 10% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. 70. The method of embodiment 67, wherein the agricultural pest applied with the agricultural composition is more sensitive to the pesticide than an agricultural pest unapplied with the agricultural composition. 71. The method of embodiment 67, wherein the pesticide is a chemical pesticide or a biological pesticide. 72. The method of embodiment 71, wherein the biological pesticide is a protein toxin. 73. The method of embodiment 67, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide. 74. The method of embodiment 67, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. INCORPORATION BY REFERENCE [00385] All references, articles, publications, patents, patent publications, and patent applications cited herein are incorporated by reference in their entireties for all purposes. However, mention of any reference, article, publication, patent, patent publication, and patent application cited herein is not, and should not, be taken as an acknowledgment or any form of suggestion that they constitute valid prior art or form part of the common general knowledge in any country in the world. 108 303137700 REFERENCES Li, X., Guo, L., Zhou, X. et al. miRNAs regulated overexpression of ryanodine receptor is involved in chlorantraniliprole resistance in Plutella xylostella (L.). Sci Rep 5, 14095 (2015) Jiang, J. H. et al. MicroRNA-281 regulates the expression of ecdysone receptor (EcR) isoform B in the silkworm, Bombyx mori. Insect Biochem Molec 43, 692–700 (2013). Markussen, M. D. K. & Kristensen, M. Low expression of nicotinic acetylcholine receptor VXEXQLW^0GĮ^^ LQ^QHRQLFRWLQRLG^ UHVLVWDQW^ VWUDLQV^RI^0XVFD^GRPHVWLFD^/^^3HVW^0DQDJ^6ci 66, 1257–1262 (2010). Bautista MA, Miyata T, Miura K, Tanaka T. RNA interference-mediated knockdown of a cytochrome P450, CYP6BG1, from the diamondback moth, Plutella xylostella, reduces larval resistance to permethrin. Insect Biochem Mol Biol. 2009 Jan;39(1):38-46. Tabashnik BE, Liu YB, Finson N, Masson L, Heckel DG. One gene in diamondback moth confers resistance to four Bacillus thuringiensis toxins. Proc Natl Acad Sci U S A. 1997;94(5):1640-1644. Huang F. Resistance of the fall armyworm, Spodoptera frugiperda, to transgenic Bacillus thuringiensis Cry1F corn in the Americas: lessons and implications for Bt corn IRM in China. Insect Science. 2020 Jun. Berger S, El Chazli Y, Babu AF, Coste AT. Azole Resistance in Aspergillus fumigatus: A Consequence of Antifungal Use in Agriculture? Front Microbiol. 2017 Jun 7;8:1024. Rupp S, Weber RW, Rieger D, Detzel P, Hahn M. Spread of Botrytis cinerea Strains with Multiple Fungicide Resistance in German Horticulture. Front Microbiol. 2017 Jan 3;7:2075. Fire, A. et al. Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature. 391, 806–811 (1998). Baulcombe, D.C. VIGS, HIGS and FIGS: small RNA silencing in the interactions of viruses or filamentous organisms with their plant hosts. Curr. Opin. Plant Biol. 26, 141–146 (2015). Wang, M. et al. Bidirectional cross-kingdom RNAi and fungal uptake of external RNAs confer plant protection. Nat. Plants 2, 16151 (2016). Xiong, F. et al. Host-induced gene silencing of BcTOR in Botrytis cinerea enhances plant resistance to grey mould. Mol. Plant Pathol. 20, 1722–1739 (2019). Islam, M.T. & Sherif, S.M. RNAi-Based Biofungicides as a promising next-generation strategy for controlling devastating gray mold diseases. Int. J. Mol. Sci. 21, 2072 (2020). 109 303137700 Wang, M., Thomas,N. & Jin, H. Cross-kingdom RNA trafficking and environmental RNAi for powerful innovative pre- and post-harvest plant protection. Curr. Opin. Plant Biol. 38, 133– 141 (2017). Mcloughlin, A.G. et al. Developing new RNA interference technologies to control fungal pathogens. Can. J. Plant Pathol. 40, 325–335 (2018). Mcloughlin, A.G. et al. Identification and application of exogenous dsRNA confers plant protection against Sclerotinia sclerotiorum and Botrytis cinerea. Sci. Rep. 8, 7320 (2018). Mitter, N. et al. Clay nanosheets for topical delivery of RNAi for sustained protection against plant viruses. Nat. Plants 3, 16207 (2017). Ferrè J. and Van Rie J. (2002) Biochemistry and genetics of insect resistance to Bacillus thuringiensis. Ann. Rev. Entomol. 47, 501-533. Avenot H.F., A. Sellam, G. Karaoglanidis, and T.J. Michailides. 2008. Characterization of mutations in the iron-sulphur subunit of succinate dehydrogenase correlating with boscalid resistance in Alternaria alternata from California pistachio. Phytopathology 98: 736-742 Avenot H.F., A. Thomas, R.D. Gitaitis R.D., D.B. Langston Jr., and K.L. Stevenson. 2011. Molecular characterization of boscalid- and penthiopyrad-resistant isolates of Didymella bryoniae and assessment of their sensitivity to fluopyram. Pest Management Science, doi: 10.1002/ps.2311. Epub 2011 Nov 10. Gudmestad N.C., S. Arabiat, J.S. Miller, and J.S. Pasche.2013. Prevalence and Impact of SDHI fungicide resistance in Alternaria solani. Plant Disease 97: 952-960. Yin, Y.N., Y.K. Kim, and C.L. Xiao. 2011. Molecular characterization of boscalid resistance in field isolates of Botrytis cinerea from apple. Phytopathology 101: 986-995. 110 303137700

Claims

CLAIMS What is claimed is: 1. An agricultural composition, comprising: a. a first minicell encapsulating a nucleic acid that is capable of inducing RNA interference in an agricultural pest, wherein the nucleic acid reduces resistance to or tolerance of a pesticide in the agricultural pest.

2. The agricultural composition of claim 1, wherein the agricultural pest is resistant to or tolerant of the pesticide.

3. The agricultural composition of claim 1 or 2, wherein the pesticide is a chemical pesticide or a biological pesticide.

4. The agricultural composition of any one of claims 1-3, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide.

5. The agricultural composition of claim 1 or 2, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird.

6. The agricultural composition of claim 1, wherein the nucleic acid is capable of recovering the agricultural pest’s sensitivity or susceptibility to the pesticide.

7. The agricultural composition of claim 1, wherein the nucleic acid is capable of altering expression of a gene responsible for pesticide resistance or tolerance.

8. The agricultural composition of claim 7, wherein the expression of the gene responsible for pesticide resistance or tolerance is downregulated.

9. The agricultural composition of claim 7, wherein the expression of the gene responsible for pesticide resistance or tolerance is upregulated.

10. The agricultural composition of claim 7, wherein the gene responsible for pesticide resistance or tolerance is an ion channel gene, a detoxification gene, a target site resistance gene, or a transporter gene.

11. The agricultural composition of claim 10, wherein the ion channel gene is a gene encoding Ryanodine receptor (RyR) or Voltage-gated sodium channel (VGSC).

12. The agricultural composition of claim 10, wherein the detoxification gene is selected from the group consisting of a gene encoding UDP-glycosyltransferase (UGT), 111 303137700 Cytochrome P450 monooxygenase, Esterase, Carboxylesterase (CarE), and Glutathione S-transferase (GST).

13. The agricultural composition of claim 10, wherein the target site resistance gene is selected from the group consisting of a gene encoding Acetylcholinesterase (AChE), Voltage-gated sodium channel (VGSC), Gamma-aminobutyric acid (GABA) receptor, Nicotinic acetylcholine receptor (nAChR), and Glutamate-gated chloride channel (GluCl).

14. The agricultural composition of claim 10, wherein the transporter gene is selected from the group consisting of a gene encoding ATP-binding cassette (ABC) transporter, Solute carrier (SLC) transporter, Major facilitator superfamily (MFS) transporter, and P-glycoprotein.

15. The agricultural composition of any one of claims 1-14, wherein the nucleic acid is capable of inducing RNA interference in at least one member from an order selected from the group consisting of: Coleoptera, Diptera, Hymenoptera, Lepidoptera, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Siphonaptera, and Trichoptera.

16. The agricultural composition of any one of claims 1-15, wherein the nucleic acid is capable of inducing RNA interference in a member of the order Lepidoptera.

17. The agricultural composition of any one of claims 1-15, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Plutella.

18. The agricultural composition of any one of claims 1-15, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Spodoptera.

19. The agricultural composition of any one of claims 1-18, wherein the nucleic acid is a RNA molecule.

20. The agricultural composition of any one of claims 1-19, wherein the nucleic acid is at least one selected from the group consisting of: a double-stranded RNA (dsRNA) a short-hairpin RNA (shRNA), a small-interfering RNA (siRNA), and a microRNA (miRNA).

21. The agricultural composition of any one of claims 1-20, wherein the nucleic acid is dsRNA.

22. The agricultural composition of any one of claims 1-20, wherein the nucleic acid is shRNA.

23. The agricultural composition of any one of claims 1-20, wherein the nucleic acid is siRNA. 112 303137700

24. The agricultural composition of any one of claims 1-20, wherein the nucleic acid is miRNA.

25. The agricultural composition of any one of claims 1-24, wherein the minicell is ribonuclease deficient.

26. The agricultural composition of any one of claims 1-25, wherein the minicell comprises at least one fusion protein.

27. The agricultural composition of any one of claims 1-26, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell.

28. The agricultural composition of any one of claims 1-27, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising at least one target cell adhesion moiety.

29. The agricultural composition of any one of claims 1-28, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising a carbohydrate binding molecule.

30. The agricultural composition of claim 3, wherein the biological pesticide is a protein toxin.

31. The agricultural composition of claim 1, further comprising: b. a second minicell encapsulating the pesticide that is capable of killing or controlling the agricultural pest.

32. The agricultural composition of any one of claims 1-31, further comprising a solid, dry, or liquid carrier.

33. The agricultural composition of claim 32, wherein said solid carrier is in a form of granule or pellet and is selected from the group consisting of: diatomaceous earth, loam, silica, alginate, clay, bentonite, vermiculite, and combinations thereof.

34. The agricultural composition of claim 32, wherein said dry carrier in a form of powder and is selected from the group consisting of: peat, wheat, bran, vermiculite, clay mineral, calcium carbonate, dolomite, gypsum, bentonite, rock phosphate, phosphorous compound, titanium dioxide, humus, talc, alginate, activated charcoal, and combinations thereof.

35. The agricultural composition of claim 32, wherein said liquid carrier is in a form of liquid or emulsion, and is selected from the group consisting of a surfactant, an emulsifier, a crop oil concentrate, a penetrant, and combinations thereof.

36. An agricultural composition, comprising: a minicell encapsulating (i) a nucleic acid capable of inducing RNA interference in an agricultural pest and (ii) a pesticide capable 113 303137700 of killing or controlling the agricultural pest, wherein the nucleic acid reduces resistance to or tolerance of the pesticide in the agricultural pest.

37. The agricultural composition of claim 36, wherein the agricultural pest is resistant to or tolerant of the pesticide.

38. The agricultural composition of claim 36, wherein the pesticide is a chemical pesticide or a biological pesticide.

39. The agricultural composition of any one of claims 36-38, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide.

40. The agricultural composition of any one of claims 36-39, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird.

41. The agricultural composition of claim 36, wherein the nucleic acid is capable of recovering the agricultural pest’s sensitivity or susceptibility to the pesticide.

42. The agricultural composition of claim 36, wherein the nucleic acid is capable of altering expression of a gene responsible for pesticide resistance or tolerance.

43. The agricultural composition of claim 42, wherein the expression of the gene responsible for pesticide resistance or tolerance is downregulated.

44. The agricultural composition of claim 42, wherein the expression of the gene responsible for pesticide resistance or tolerance is upregulated.

45. The agricultural composition of claim 42, wherein the gene responsible for pesticide resistance or tolerance is an ion channel gene, a detoxification gene, a target site resistance gene, or a transporter gene.

46. The agricultural composition of claim 45, wherein the ion channel gene is a gene encoding Ryanodine receptor (RyR) or Voltage-gated sodium channel (VGSC).

47. The agricultural composition of claim 45, wherein the detoxification gene is selected from the group consisting of a gene encoding UDP-glycosyltransferase (UGT), Cytochrome P450 monooxygenase, Esterase, Carboxylesterase (CarE), and Glutathione S-transferase (GST).

48. The agricultural composition of claim 45, wherein the target site resistance gene is selected from the group consisting of a gene encoding Acetylcholinesterase (AChE), Voltage-gated sodium channel (VGSC), Gamma-aminobutyric acid (GABA) receptor, 114 303137700 Nicotinic acetylcholine receptor (nAChR), and Glutamate-gated chloride channel (GluCl).

49. The agricultural composition of claim 45, wherein the transporter gene is selected from the group consisting of a gene encoding ATP-binding cassette (ABC) transporter, Solute carrier (SLC) transporter, Major facilitator superfamily (MFS) transporter, and P-glycoprotein.

50. The agricultural composition of any one of claims 36-49, wherein the nucleic acid is capable of inducing RNA interference in at least one member from an order selected from the group consisting of: Coleoptera, Diptera, Hymenoptera, Lepidoptera, Homoptera, Hemiptera, Orthoptera, Thysanoptera, Dermaptera, Siphonaptera, and Trichoptera.

51. The agricultural composition of any one of claims 36-50, wherein the nucleic acid is capable of inducing RNA interference in a member of the order Lepidoptera.

52. The agricultural composition of any one of claims 36-50, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Plutella.

53. The agricultural composition of any one of claims 36-50, wherein the nucleic acid is capable of inducing RNA interference in a member of the genus Spodoptera.

54. The agricultural composition of any one of claims 36-53, wherein the nucleic acid is a RNA molecule.

55. The agricultural composition of any one of claims 36-54, wherein the nucleic acid is at least one selected from the group consisting of: a double-stranded RNA (dsRNA), a short-hairpin RNA (shRNA), a small-interfering RNA (siRNA), and a microRNA (miRNA).

56. The agricultural composition of any one of claims 36-55, wherein the nucleic acid is dsRNA.

57. The agricultural composition of any one of claims 36-55, wherein the nucleic acid is shRNA.

58. The agricultural composition of any one of claims 36-55, wherein the nucleic acid is siRNA.

59. The agricultural composition of any one of claims 36-55, wherein the nucleic acid is miRNA.

60. The agricultural composition of any one of claims 36-59, wherein the minicell is ribonuclease deficient. 115 303137700

61. The agricultural composition of any one of claims 36-60, wherein the minicell comprises at least one fusion protein.

62. The agricultural composition of any one of claims 36-61, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell.

63. The agricultural composition of any one of claims 36-62, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising at least one target cell adhesion moiety.

64. The agricultural composition of any one of claims 36-63, wherein the minicell comprises at least one fusion protein expressed on the surface of the minicell, said fusion protein comprising a carbohydrate binding molecule.

65. The agricultural composition of claim 38, wherein the biological pesticide is a protein toxin.

66. The agricultural composition of any one of claims 36-65, further comprising a solid, dry, or liquid carrier.

67. The agricultural composition of claim 66, wherein said solid carrier is in a form of granule or pellet and is selected from the group consisting of: diatomaceous earth, loam, silica, alginate, clay, bentonite, vermiculite, and combinations thereof.

68. The agricultural composition of claim 66, wherein said dry carrier in a form of powder and is selected from the group consisting of: peat, wheat, bran, vermiculite, clay mineral, calcium carbonate, dolomite, gypsum, bentonite, rock phosphate, phosphorous compound, titanium dioxide, humus, talc, alginate, activated charcoal, and combinations thereof.

69. The agricultural composition of claim 66, wherein said liquid carrier is in a form of liquid or emulsion, and is selected from the group consisting of a surfactant, an emulsifier, a crop oil concentrate, a penetrant, and combinations thereof.

70. A method of reducing or suppressing pesticide resistance in an agricultural pest, the method comprising: applying an agricultural composition of claim 1 or 36 to an agricultural pest, wherein resistance to a pesticide in the agricultural pest is reduced or suppressed after the application of the agricultural composition.

71. The method of claim 70, wherein the agricultural pest is resistant to or tolerant of the pesticide.

72. The method of claim 70, wherein the resistance to the pesticide is reduced at least 10% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition. 116 303137700

73. The method of claim 70, wherein the pesticide is a chemical pesticide or a biological pesticide.

74. The method of claim 73, wherein the biological pesticide is a protein toxin.

75. The method of claim 70, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide.

76. The method of claim 70, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird.

77. A method of restoring susceptibility of an agricultural pest to a pesticide, the method comprising: applying an agricultural composition of claim 1 or 36 to an agricultural pest, wherein the agricultural pest is restored to be susceptible to a pesticide after the application of the agricultural composition.

78. The method of claim 77, wherein the agricultural pest is resistant to or tolerant of the pesticide.

79. The method of claim 77, wherein the susceptibility to the pesticide is restored at least 10% in the agricultural pest applied with the agricultural composition when comparing to an agricultural pest unapplied with the agricultural composition.

80. The method of claim 77, wherein the agricultural pest applied with the agricultural composition is more sensitive to the pesticide than an agricultural pest unapplied with the agricultural composition.

81. The method of claim 77, wherein the pesticide is a chemical pesticide or a biological pesticide.

82. The method of claim 81, wherein the biological pesticide is a protein toxin.

83. The method of claim 77, wherein the pesticide is selected from the group consisting of an insecticide, a herbicide, a fungicide, an algaecide, a bactericide, a rodenticide, a larvicide, a repellent, a virucide, an ovicide, an acaricide, a nematicide, a molluscicide, a pediculicide, a piscicide, and an avicide.

84. The method of claim 77, wherein the agricultural pest is selected from the group consisting of insect, weed, fungus, algae, bacterium, rodent, larvae, virus, mite, tick, nematode, mollusca, lice, fish, and bird. 117 303137700