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WO2023070165A1 - Procédé de production d'une réponse biologique - Google Patents

Procédé de production d'une réponse biologique Download PDF

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Publication number
WO2023070165A1
WO2023070165A1 PCT/AU2022/051302 AU2022051302W WO2023070165A1 WO 2023070165 A1 WO2023070165 A1 WO 2023070165A1 AU 2022051302 W AU2022051302 W AU 2022051302W WO 2023070165 A1 WO2023070165 A1 WO 2023070165A1
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Prior art keywords
plant
bioactive
extract
approximately
terpene
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PCT/AU2022/051302
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English (en)
Inventor
Richard Woods
James Walker
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Supercritical Pty Ltd
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Supercritical Pty Ltd
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Priority claimed from AU2021903450A external-priority patent/AU2021903450A0/en
Application filed by Supercritical Pty Ltd filed Critical Supercritical Pty Ltd
Publication of WO2023070165A1 publication Critical patent/WO2023070165A1/fr
Anticipated expiration legal-status Critical
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Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N27/00—Biocides, pest repellants or attractants, or plant growth regulators containing hydrocarbons
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H3/00—Processes for modifying phenotypes, e.g. symbiosis with bacteria
    • A01H3/04—Processes for modifying phenotypes, e.g. symbiosis with bacteria by treatment with chemicals
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/02—Flowers
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H5/00—Angiosperms, i.e. flowering plants, characterised by their plant parts; Angiosperms characterised otherwise than by their botanic taxonomy
    • A01H5/12—Leaves
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H6/00—Angiosperms, i.e. flowering plants, characterised by their botanic taxonomy
    • A01H6/28—Cannabaceae, e.g. cannabis
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/02—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests containing liquids as carriers, diluents or solvents
    • A01N25/04—Dispersions, emulsions, suspoemulsions, suspension concentrates or gels
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N49/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds containing the group, wherein m+n>=1, both X together may also mean —Y— or a direct carbon-to-carbon bond, and the carbon atoms marked with an asterisk are not part of any ring system other than that which may be formed by the atoms X, the carbon atoms in square brackets being part of any acyclic or cyclic structure, or the group, wherein A means a carbon atom or Y, n>=0, and not more than one of these carbon atoms being a member of the same ring system, e.g. juvenile insect hormones or mimics thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P3/00—Fungicides
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P7/00—Arthropodicides
    • A01P7/02—Acaricides
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P7/00—Arthropodicides
    • A01P7/04—Insecticides

Definitions

  • this invention relates to uses of a bioactive composition, preferably an oil extract of cannabis, which has useful bioactive properties.
  • the invention concerns a bioactive composition in the form of an oil extract of cannabis, capable of providing useful effects against biological organisms such as pesticidal effects or allelopathic effects.
  • Secondary plant metabolites are classified according to their chemical structures into several broad classes which include phenolics, alkaloids, saponins, terpenes, lipids and carbohydrates. Of these, the terpenes appear to be the most chemically and functionally diverse. Terpenes are naturally occurring hydrocarbons produced by a wide variety of plants and are the primary constituents of essential oils. The classification of terpenes is based on five-carbon (isoprene) units as their building blocks.
  • terpenes include hemiterpenes (C5), monoterpenes (CIO), sesquiterpenes (C15), diterpenes (C20), sesterterpenes (C25), triterpenes (C30), and polyterpenes (>C30).
  • hempbis refers to the plants of the species Cannabis Sativa L. Within that species are three types of cannabis plants: Cannabis Sativa, Cannabis Indica and Cannabis Ruderalis. ‘Hemp’ and cannabis are the same species of plant except that hemp produces less of the psychoactive chemical Delta-9-tetrahydrocannabinol (THC).
  • THC psychoactive chemical Delta-9-tetrahydrocannabinol
  • terpene profile Around 150 different terpene compounds have been identified in the cannabis plant, and each cannabis cultivar is thought to have a unique constitution and ratio of terpenes (terpene profile).
  • Myrcene is generally the most abundant terpene in modern commercial cannabis. On average, myrcene is thought to represent over 20% of the terpene profile in most cultivars although individual cultivars do vary widely in their terpene content.
  • Table 1 below shows six cannabis terpenes generally recognised as the most common among cultivars and various biological properties that have been associated with that terpene.
  • Allelopathy is a process whereby an organism (usually a plant) produces a negative or positive effect on another organism (usually another plant, but sometimes an insect), by producing a chemical compound (allelochemical) which interferes with the biology of the other organism.
  • a chemical compound allelochemical which interferes with the biology of the other organism.
  • a switch from stimulatory effects to inhibitory activity with increasing concentrations was observed (suggesting a response threshold). It is thought that the production of compounds that have an inhibitory effect on other organisms enable the producing organism to limit growth of competitors and thus facilitate access to resources.
  • VOC volatile organic compounds
  • the VOC’s includes the terpenes and other volatile plant compounds. These compounds are given off the plant in a gaseous form (or fragrance) in normal daytime temperatures when the plant is at that stage of its life cycle. Nearby plants and insects are able to ‘sense’ and react to these VOC’s. This is known as plant volatile-mediated plant-to-plant communication. Recent research has shown how the genetic mechanisms of the receiving plant adjust as various genes are up or down regulated in response to the VOC’s (Nagashima (2018)).
  • Examples of this include research in some vegetable species which have been found to release certain volatiles in response to attack by insect herbivores and pathogens. These induced volatiles can attract the natural enemies of insect herbivores, repel other insect herbivores and increase the resistance of tissues in both the host plant and neighbouring plants (references cited in Kang et.al 2018). Furthermore, intermittent exposure to the volatiles emitted from artificially damaged Arabidopsis has been shown to induce defensive responses in undamaged neighbouring plants (Shiojiri et al., (2012) in Kang (2016)).
  • the present invention in one broad form, relates to methods of producing a biological response in a plant from the Cannabaceae family which involves administering a cannabis extract to the plant.
  • a method of producing a biological response in a plant from the Cannabaceae family comprising the step of administering a bioactive oil extract to the plant to thereby produce the biological response, wherein the bioactive oil extract is a cannabis extract.
  • the present inventors have found that by administering a cannabis derived bioactive oil extract to plants of the Cannabaceae family, it is possible to produce a range of beneficial biological responses in the subject plant, such as increased resistance to pest infestation, increased resistance to mould infection, increased resistance to bacterial, fungal or viral infection, and increased biomass production. This result was unexpected, as in the method an extract from cannabis is applied to plants of the same family. To date, there has been relatively little research directed towards the potential of positive allelopathic effects, particularly those that may be found when an extract of one plant is applied to a phylo genetically similar plant species.
  • the cannabis extract comprises, consists essentially of, or consists of the following concentrations of monoterpenes:
  • Terpene Profile 1 approximately 0.3-50.1 % Pinene; approximately 0.1-10.9 % Limonene; and approximately 0.2-39.0 % Ocimene.
  • Terpene Profile 1 comprises, consists essentially of, or consists of the following concentrations of monoterpenes: approximately 19-31 % Pinene; approximately 4-6 % Limonene; approximately 18-21 % Ocimene.
  • Terpene profile 1 comprises, consists essentially of, or consists of Pinene; Limonene; and Ocimene in a ratio of about 5:1:4.
  • the ratio of Pinene: limonene in Terpene Profile 1 may be between about 3:1 and about 6:1.
  • the ratio of Ocimene: limonene in Terpene Profile 1 may be between about 3:1 and about 5:1.
  • Terpene Profile 1 comprises, consists essentially of, or consists of the following concentrations of monoterpenes: approximately 0.2-35.2 % a- Pinene; approximately 0.1-14.9 % P -Pinene; approximately 0.1-10.9 % D-Limonene; approximately 0.0- 1.5 % Trans-Ocimene; and approximately 0.2-37.5 % Cis-Ocimene.
  • Terpene Profile 1 comprises, consists essentially of, or consists of a-Pinene; P-Pinene; D-Limonene; Trans-Ocimene; and Cis-Ocimene in a ratio of about 22:9:7:1:24.
  • Terpene Profile 1 comprises, consists essentially of, or consists of a-Pinene; P-Pinene; D-Limonene; Trans-Ocimene; and Cis-Ocimene in a ratio of about 19-27:9-11:6-8:1:22-29.
  • the ratio of a-Pinene:Trans-Ocimene in Terpene Profile 1 may be between about 19:1 and about 27:1.
  • the ratio of P-Pinene:Trans-Ocimene in Terpene Profile 1 may be between about 9:1 and about 11:1.
  • the ratio of D-Limonene:Trans-Ocimene in Terpene Profile 1 may be between about 6:1 and about 8:1.
  • the ratio of Cis-Ocimene:Trans-Ocimene in Terpene Profile 1 may be between about 22: 1 and about 29:1.
  • the cannabis extract comprises, consists essentially of, or consists of the following concentrations of monoterpenes:
  • Terpene Profile 2 approximately 4.8-45.6 % Pinene; approximately 1.0-9.0 % Limonene; approximately 3.7-35.5 % Ocimene; and approximately 1.1-9.9 % P -Myrcene.
  • Terpene Profile 2 comprises, consists essentially of, or consists of the following concentrations of monoterpenes: approximately 19-31 % Pinene; approximately 4-6 % Limonene; approximately 4-6 % P-Myrcene; and approximately 18-21 % Ocimene.
  • Terpene profile 2 comprises, consists essentially of, or consists of Pinene; Limonene; P-Myrcene and Ocimene in a ratio of about 5: 1:1:4.
  • the ratio of Pinene: limonene in Terpene Profile 2 may be between about 3:1 and about 6:1.
  • the ratio of P-Myrcene:limonene in Terpene Profile 2 may be between about 0.5:1 and about 1.5:1.
  • the ratio of Ocimene: limonene in Terpene Profile 2 may be between about 3:1 and about 5:1.
  • Terpene Profile 2 comprises, consists essentially of, or consists of the following concentrations of monoterpenes: approximately 3.4-32.0 % a- Pinene; approximately 0.1-13.6 % P -Pinene; approximately 1.1-9.9% D-Limonene; approximately 0.1- 1.4 % Trans-Ocimene; approximately 3.6-34.1 % Cis-Ocimene; and approximately 1.0-9.0 % P-Myrcene.
  • Terpene Profile 2 comprises, consists essentially of, or consists of a-Pinene; P-Pinene; D-Limonene; P-Myrcene; Trans-Ocimene; and Cis-Ocimene in a ratio of 22:9:7:6: 1 :24.
  • Terpene Profile 2 comprises, consists essentially of, or consists of a-Pinene; P-Pinene; D-Limonene; P-Myrcene; Trans-Ocimene; and Cis-Ocimene in a ratio of 19-27:9-11:6-8:6-7:1:22-29
  • the ratio of a-Pinene:Trans-Ocimene in Terpene Profile 2 may be between about 19:1 and about 27:1.
  • the ratio of P-Pinene:Trans-Ocimene in Terpene Profile 2 may be between about 9:1 and about 11:1.
  • the ratio of D-Limonene:Trans-Ocimene in Terpene Profile 2 may be between about 6:1 and about 8:1.
  • the ratio of P-Myrcene:Trans-Ocimene in Terpene Profile 2 may be between about 6:1 and about 7:1.
  • the ratio of Cis-Ocimene:Trans-Ocimene in Terpene Profile 2 may be between about 22: 1 and about 29:1.
  • the cannabis extract is a liquid.
  • the cannabis extract is an essential oil extract.
  • the cannabis extract is a solid.
  • the cannabis extract is in the form of a dry powder, pellet, granules, tablets or flakes.
  • the cannabis extract is administered as part of a composition, wherein the composition comprises one or more excipients.
  • the one or more excipients are selected from the group consisting of: drying agents; solvent; adhesive; aqueous or oily diluent; carrier; base; buffer; pH adjuster; bittering agent (i.e. foul-tasting agent); suspending agent; thickening agent; rheology modifier; gelling agent; viscosity increasing agent; antifreeze, emulsifier; emollient; stabilising agent; dispersing agent/dispersant; solubiliser; fragrance; preservative; surfactant; textural modifier; foaming agent; anti-foaming agent; colourant; propellant; refrigerant; and, waterproofing agent.
  • the other excipients further comprise a fertiliser.
  • composition may be an emulsion.
  • the emulsion is an oil-in-water emulsion. In another embodiment, the emulsion is a water-in-oil emulsion.
  • the composition is a colloid, suspension, admixture, solution or mixture.
  • the cannabis extract is an extract of cannabis flowers.
  • the cannabis extract is an extract of cannabis flowers, panicles, stem, or upper leaves, or a combination thereof.
  • the cannabis extract is an extract of a single cannabis variety. In other embodiments, the cannabis extract is an extract of more than one cannabis variety.
  • the cannabis extract may administered during the vegetative growth stage of development of the plant.
  • the cannabis extract is administered during the seed, seedling or vegetative stages of development of the plant, or a combination thereof. In other embodiments, the cannabis extract is administered to plant cuttings, either before or after root development.
  • the biological response is increased biomass production.
  • the biological response is increased resistance to pest infestation.
  • the pest may be selected from the group consisting of: insect and arachnid; especially from the group consisting of: mosquito, fruit fly, spider mite and aphid.
  • the pest is selected from the group consisting of: caterpillar, scale, mosquito, fly, aphid, mite, spider, cockroach, weevil and rootworm.
  • the biological response is increased resistance to bacterial, fungal or viral infection.
  • the fungal infection is a mould.
  • the biological response is stimulating the production of plant defence compounds within the plant.
  • the production of plant defence compounds provides increased resistance to pest infestation.
  • the plant is a cannabis plant.
  • the plant is a variety of hops (Humulus lupulus) plant.
  • the cannabis extract is administered to the leaves of the plant.
  • the cannabis extract is administered to the roots, foliage, flower head or seed, or a combination thereof, of the plant.
  • the bioactive oil extract is a hemp extract.
  • the method may be a prophylactic method.
  • the method may be a preventative method.
  • the method of producing a biological response is preferably a method of producing a beneficial biological response.
  • a method of controlling one or more properties of a plant species in the Cannabaceae family comprising the step of administering to the plant a bioactive oil extract comprising the following concentrations of monoterpenes:
  • Terpene Profile 1 approximately 0.3-50.1 % Pinene; approximately 0.1-10.9 % Limonene; and approximately 0.2-39.0 % Ocimene, or
  • Terpene Profile 2 approximately 4.8-45.6 % Pinene; approximately 1.0-9.0 % Limonene; approximately 3.7-35.5 % Ocimene; and approximately 1.1-9.9 % [3-Myrcene, and wherein the one or more properties are selected from the group consisting of: stimulating the production of plant defence compounds within the plant; triggering anti-microbial defence mechanisms within the plant; activating anti-viral defence mechanisms within the plant; activating anti-bacterial defence mechanisms within the plant; eliciting plant disease resistance; enhancing plant disease resistance; promoting plant growth; increasing biomass production within the plant; increasing plant uniformity; or, use as a herbicide.
  • the one or more properties may be stimulating the production of plant defence compounds within the plant.
  • the production of plant defence compounds may provide increased resistance to pest infestation.
  • a method of controlling one or more properties in a plant from the Cannabaceae family comprising the step of administering a bioactive oil extract to the plant to thereby control the one or more properties, wherein the bioactive oil extract is a cannabis extract.
  • the cannabis extract is a fertiliser.
  • the method of producing a biological response is a method of eliciting a biological response.
  • the method of producing a biological response is method of producing an allelopathic effect.
  • a method of increasing biomass production in a plant from the Cannabaceae family comprising the step of administering a bioactive oil extract to the plant to thereby increase biomass production, wherein the bioactive oil extract is a cannabis extract.
  • a method increasing resistance to pest infestation in a plant from the Cannabaceae family comprising the step of administering a bioactive oil extract to the plant to thereby increase resistance to pest infestation, wherein the bioactive oil extract is a cannabis extract.
  • a method increasing resistance to fungal infection in a plant from the Cannabaceae family comprising the step of administering a bioactive oil extract to the plant to thereby increase resistance to fungal infection, wherein the bioactive oil extract is a cannabis extract.
  • the present inventors have produced a bioactive terpene composition based on an oil extract of cannabis plants which has bioactive properties, and have further characterised a terpene profile of that extract that provides useful effects against biological organisms, such as pesticidal effects or allelopathic effects.
  • a cannabis-based bioactive terpene composition comprising the following concentrations of monoterpenes:
  • a bioactive terpene composition comprising a bioactive oil extract prepared from cannabis plants, wherein the composition comprises the following concentrations of monoterpenes:
  • % means percentage volume or percentage weight. Each of those ranges includes all 0.01% incremental values between the upper and lower concentration limits.
  • bioactive oil extract based on or prepared from cannabis plants, wherein the bioactive oil extract comprises the following concentration of monoterpenes:
  • a method of preparing the bioactive terpene composition of the seventh or eighth aspect, or the bioactive oil extract of the ninth aspect comprising the step of:
  • a preparation step comprising preparing a bioactive oil extract from cannabis plants.
  • a cannabis-based bioactive oil extract prepared using the method according to the tenth aspect.
  • a formulation comprising the bioactive terpene composition of the seventh or eighth aspect, or the bioactive oil extract of the ninth or tenth aspect, and, optionally, at least one additional ingredient.
  • a fourteenth aspect of the present invention there is provided a method of controlling a biological organism, said method comprising the step of treating the biological organism with or exposing the biological organism to:
  • Plant material derived or obtained from the hemp variety can comprise flowers, panicles, stems, roots or leaves. Plant material derived or obtained from the hemp variety can comprise live plant material that can be propagated, such as cuttings or seeds.
  • the bioactive terpene composition or bioactive oil extract is based on or prepared from cannabis plants of the species Cannabis Sativa L, preferably an essential oil extract. Any suitable type or types of cannabis plants can be used, including Cannabis Sativa, Cannabis Indica and/or Cannabis Ruderalis.
  • the cannabis plants can have psychoactive properties.
  • the cannabis plants can be hemp, and thus lack psychoactive properties.
  • the bioactive terpene composition or bioactive oil extract is based on or prepared from hemp, preferably an essential oil extract. In some embodiments, the bioactive terpene composition or bioactive oil extract is based on or prepared from a single cannabis plant cultivar, variety or chemovar, preferably an essential oil extract. In some embodiments, the bioactive terpene composition or bioactive oil extract is based on or prepared from more than one cannabis plant cultivar, variety or chemovar, including two, three, four or more cannabis plant cultivars, varieties or chemovars, preferably an essential oil extract.
  • the bioactive terpene composition or bioactive oil extract preferably further comprises P-Myrcene, as shown in Terpene Profile 2.
  • the monoterpenes can each comprise one or more different isomeric forms.
  • the pinene terpene can comprise a-pinene and P-pinene.
  • the limonene terpene can comprise D-limonene.
  • the ocimene terpene can comprise trans-ocimene and cis-ocimene.
  • the myrcene terpene can comprise P-myrcene.
  • the bioactive terpene composition or bioactive oil extract is further prepared by the addition of at least one oil additive, such as at least one monoterpene and/or at least one oil extract (preferably an essential oil extract), such that a terpene profile of interest is produced.
  • the oil additive is an oil extract from non-cannabis plants, preferably an essential oil extract.
  • the oil additive is an oil extract from plants in the family Cannabaceae, preferably an essential oil extract.
  • the oil additive is an oil extract from plants not in the family Cannabaceae, preferably an essential oil extract.
  • the oil additive is a naturally-derived or synthetic monoterpene. If naturally-derived, it can be produced from cannabis plants, non-cannabis plants, plants in the family Cannabaceae, or plants not in the family Cannabaceae.
  • the cannabis-based bioactive terpene composition comprises the bioactive oil extract according to the ninth aspect of the invention, but this need not be the case.
  • the bioactive oil extract preferably an essential oil extract, can be prepared in any suitable way.
  • the bioactive oil extract is prepared by alcohol extraction, such as ethanol extraction.
  • the bioactive oil extract is prepared by oil extraction, for example, using olive oil or coconut oil.
  • the bioactive oil extract is prepared by extraction using a hydrocarbon such as butane.
  • the bioactive oil extract is prepared by distillation. In some embodiments, the bioactive oil extract is prepared by steam distillation and/or hydrodistillation. Steam distillation and hydrodistillation procedures are well known in the art, and examples can be found in the following reference, the entire contents of which are incorporated herein by reference: Jamil et al. (2016).
  • the bioactive oil extract is prepared by using carbon dioxide (CO2) extraction technology, such as subcritical or supercritical CO2 extraction.
  • CO2 carbon dioxide
  • Carbon dioxide extraction procedures are well known in the art, and examples can be found in the following reference, the entire contents of which are incorporated herein by reference: Naz et al. (2017).
  • the bioactive oil extract is prepared by distillation, preferably steam distillation, of fresh plant material or raw, frozen plant material. Any suitable part or parts of the plant can be used. Any suitable part or parts of the cannabis plant can be used, including the flowers, panicles, stem, and upper leaves of the cannabis/hemp plant. In some embodiments, the bioactive oil extract is prepared by steam distillation of cannabis flowers, preferably an essential oil extract.
  • the bioactive terpene composition or bioactive oil extract can be in a concentrated form, undiluted/neat form or diluted form.
  • one or more of the monoterpenes of the bioactive terpene composition can be prepared in any suitable way.
  • the terpenes are prepared by alcohol extraction, such as ethanol extraction.
  • the terpenes are prepared by oil extraction, for example, using olive oil or coconut oil.
  • the terpenes are prepared by extraction using a hydrocarbon such as butane.
  • the terpenes are prepared by distillation. In some embodiments, the terpenes are prepared by steam distillation and/or hydrodistillation.
  • the terpenes are prepared by using carbon dioxide (CO2) extraction technology, such as subcritical or supercritical CO2 extraction.
  • CO2 carbon dioxide
  • the terpenes are prepared by distillation, preferably steam distillation, of fresh plant material or raw, frozen plant material. Any suitable part or parts of the plant can be used. Any suitable part or parts of the cannabis plant can be used, including the flowers, panicles, stem, and upper leaves of the cannabis/hemp plant.
  • the terpenes are prepared by steam distillation of cannabis flowers, preferably an essential oil extract.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety M183, as seen in Tables 2 or 6B. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of hemp variety M183, as seen in Tables 2 or 6B.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety M168, as seen in Tables 2 or 6D. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of hemp variety M168, as seen in Tables 2 or 6D.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile is produced by blending oil extracts of two or more hemp varieties together.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the average % terpene concentration of M183 and M168 as seen in Table 2. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of the average % terpene concentration of M183 and M168 as seen in Table 2.
  • Terpene Profile 1 comprises any one or more of:
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety Ml 83, as seen in Table 3. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of hemp variety M183, as seen in Table 3.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety M168, as seen in Table 3. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of the essential oil constituents of hemp variety M168, as seen in Table 3.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the average % terpene concentration of M183 and M168 as seen in Table 3. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of the average % terpene concentration of M183 and M168 as seen in Table 3.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety M183, as seen in Table 4. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of the essential oil constituents of hemp variety M183, as seen in Table 4.
  • bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety M168, as seen in Table 4.
  • bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of hemp variety M168, as seen in Table 4.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile is produced by blending the oil extracts of hemp varieties together.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the average % terpene concentration of M183 and M168 as seen in Table 4.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of the average % terpene concentration of M183 and M168 as seen in Table 4.
  • Terpene Profile 2 comprises any one or more of:
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety M183, as seen in Table 5. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of hemp variety M183, as seen in Table 5.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety M168, as seen in Table 5. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of hemp variety M168, as seen in Table 5.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the average % terpene concentration of M183 and M168 as seen in Table 5. In some embodiments, the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of the average % terpene concentration of M183 and M168 as seen in Table 5. [179] Table 5 (Terpene Profile 2)
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile can have monoterpene concentration ranges substantially as shown in Table 7B. Each of those ranges includes all 0.01% incremental values between the upper and lower concentration limits.
  • the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile can have terpene concentration ranges substantially as shown in Tables 4, 5 or 7A. Each of those ranges includes all 0.01% incremental values between the upper and lower concentration limits.
  • the bioactive terpene composition or formulation can be of any suitable form.
  • the bioactive terpene composition or formulation can be, for example, in the form of a liquid, solution, gel, creme, emulsion, paste, film, powder, foam, suspension, gas, vapour or aerosol.
  • the bioactive oil extract can be microencapsulated using any suitable microencapsulation process.
  • the bioactive terpene composition or formulation can comprise any suitable quantity of bioactive oil extract.
  • the ratio of bioactive oil extract to all other ingredients of the bioactive terpene composition or formulation ranges between about 1:1 to about 1:1000, and any numerical value or subrange there between.
  • the bioactive terpene composition or formulation comprises approximately 200 g/L bioactive oil extract.
  • the bioactive terpene composition or formulation can include one or more of the following types of ingredients: solvent; adhesive; aqueous or oily diluent; carrier; excipient; base; buffer; pH adjuster; bittering agent (i.e. foul-tasting agent); suspending agent; thickening agent; rheology modifier; gelling agent; viscosity increasing agent; antifreeze, emulsifier; emollient; stabilising agent; dispersing agent/dispersant; solubiliser; fragrance; preservative; surfactant; textural modifier; foaming agent; anti-foaming agent; colourant; propellant; refrigerant; and, waterproofing agent.
  • the bioactive terpene composition or formulation can include, for example: oleic acid or methyl ester as solvent; polymeric surfactant, oxirane, 2- methyl-, polymer with oxirane, or monobutyl ether as emulsifier; propylene glycol or propane- 1,2-diol as antifreeze; xanthum gum as rheology modifier; and, polydimethysiloxane aqueous emulsion as an anti-foaming agent.
  • the bioactive terpene composition or formulation can include, for example, alcohol or water as an excipient or carrier.
  • Suitable oily or aqueous bases, carriers, diluents and excipients are inert and include, for example: bacteriostatic saline (saline containing benzyl alcohol), cetomacrogol, cetyl alcohol, glycerine, lanolin, petrolatum based creams, gels, hydrogels, saline, short chain alcohols and glycols (e.g. ethyl alcohol and propylene glycol), and water.
  • Either water in oil or oil in water emulsions can be used.
  • suitable surfactants and emulsifying agents include: vitamin E, non-ionic ethoxylated and nonethoxylated surfactants, abietic acid, almond oil PEG, beeswax, butylglucoside caprate, Cis- C36 acid glycol ester, C9-C15 alkyl phosphate, caprylic/capric triglyceride PEG-4 esters, cetomacrogol, ceteareth-7, cetereth-20, cetyl phosphate, cetyl stearyl alcohol, com oil PEG esters, DEA-cetyl phosphate, dextrin laurate, dilaureth-7 citrate, dimyristyl phosphate, glycereth-17 cocoate, glyceryl erucate, glycerol, glyceryl laurate, G.M.S.
  • PEG esters isosteareth-11 carboxylic acid, lecithin, lysolecithin, nonoxynol-9, octyldodeceth-20, palm glyceride, PEG diisostearate, PEG stearamine, poloxamines, polyglyceryls, potassium linoleate, PPGs, raffinose myristate, sodium caproyl lactylate, sodium caprylate, sodium cocoate, sodium isostearate, sodium tocopheryl phosphate, steareths, TEA-C12-C13 pareth-3 sulfate, tri-Cn-Cis pareth-6 phosphate, and trideceths.
  • the bioactive terpene composition or formulation can comprise one or more types of preservative.
  • a suitable preservative for example, can be: benzalkonium chloride, benzoic acid, benzothonium chloride, benzyl alcohol, 2-bromo-2-nitropropane-l,3-diol, bronopol, butylated hydroxyanisole, butylated hydroxytoluene, butyl paraben, chlorophene, chlorphenesin, diazolidinyl urea, DMDM hydantoin, ethyl paraben, formaldehyde-releasing preservative, hydroquinone, iodopropynyl butylcarbamate, imidazolidinyl urea, methyldibromo glutaronitrile, methylhydroquinone, methylisothiazolinone, methyl paraben, nitrosamines, o-cymen-5-ol,
  • the bioactive terpene composition or formulation can include a colourant so that its application can be verified visually.
  • the colourant can be a pigment and/or dye.
  • the bioactive terpene composition or formulation can comprise one or more of the following adhesives, thickening agents, gelling agents, rheology-modifiers and/or viscosity increasing agents: acrylamides copolymer, agarose, amylopectin, calcium alginate, calcium carboxymethyl cellulose, carbomer, carboxymethyl chitin, castor oil derivatives, cellulose gum, cellulosic preparation, cetyl alcohol, cetostearyl alcohol, dextrin, gelatin, hydroxy cellulose, hydroxyethylcellulose, hydroxypropylcellulose, hydroxpropyl starch, inert sugar, magnesium alginate, methylcellulose, microcrystalline cellulose, pectin, PEG's, polyacrylic acid, polymethacrylic acid, polyvinyl alcohol, quatemium ammonium compound of bentonite or zinc stearate, sorbitol, PPG's, sodium acrylates copolymer, sodium carrageenan,
  • the bioactive terpene composition, formulation or bioactive extract can be applied or used in any suitable form.
  • the bioactive terpene composition, formulation or bioactive extract can be applied or used in a liquid form or other free-flowing form.
  • the bioactive terpene composition, formulation or bioactive extract can be applied or used as a spray-on liquid, spray-on gel, foam or aerosol.
  • the bioactive terpene composition, formulation or bioactive extract can be applied as a gel by hand, or squeezed from a tube.
  • the bioactive terpene composition, the bioactive oil extract, or the formulation aspect can be used for controlling a biological organism. Any suitable type of biological organism can be controlled and this can be achieved in any suitable way.
  • control can mean, for example, repelling, killing or otherwise changing the normal behaviour of the biological organism.
  • the biological organism may be an insect, mite, spider or the like having a thin cuticle, or bacteria, fungus or virus.
  • control can mean, for example, allelopathically effecting a trait of the biological organism in a positive or negative way.
  • the biological organism may be a plant.
  • the bioactive terpene composition, bioactive oil extract, or formulation is formulated for use as a pest repellant, pesticide, insect repellant, insecticide, mite repellant, miticide, arachnid repellant, acaricide, anti-fungal, fungicide, anti-bacterial, bactericide, anti-viral, viricide, and/or for plant allelopathy (positive or negative effect).
  • the bioactive terpene composition, bioactive oil extract, or formulation is capable of controlling an insect, mite, spider, or a plant. In some embodiments, the bioactive terpene composition, bioactive oil extract, or formulation is capable of controlling mosquitoes or mosquito larvae, flies or fly larvae, aphids, mites, spiders, cockroaches, weevils or rootworms.
  • the bioactive terpene composition, bioactive oil extract, or formulation is capable of controlling a fungus, bacteria or virus.
  • the bioactive terpene composition, bioactive oil extract, or formulation is capable of controlling one or more properties of a plant, either positively or negatively.
  • the bioactive terpene composition, bioactive oil extract, or formulation is capable of controlling one or more properties of plant species in the Cannabaceae family, such as cannabis, hemp or hops varieties.
  • the bioactive terpene composition, bioactive oil extract, or formulation is capable of: stimulating the production of plant defence compounds within a plant; triggering anti-microbial defence mechanisms within a plant; activating anti-viral defence mechanisms within a plant; activating anti-bacterial defence mechanisms within a plant; eliciting plant disease resistance; enhancing plant disease resistance; promoting plant growth; increasing biomass production within a plant; increasing plant uniformity; or, use as a herbicide.
  • the bioactive terpene composition, bioactive oil extract, or formulation is capable of producing an allelochemical capable of interfering with the biology of the biological organism.
  • the step of controlling or treating the biological organism can be carried out in any suitable way.
  • the biological organism can be sprayed with the bioactive terpene composition, bioactive oil extract, or formulation, or the bioactive terpene composition, bioactive oil extract, or formulation can be applied to a surface, substrate (eg. soil) or plant surface (eg. roots, foliage, flower head, seed) on which may be found the biological organism or which may be in close proximity to the biological organism, or the bioactive terpene composition, bioactive oil extract, or formulation can be dispersed in the biological organism’s immediate vicinity or environment (eg. dispersed in the air).
  • substrate eg. soil
  • plant surface eg. roots, foliage, flower head, seed
  • the plant may be sprayed with the bioactive terpene composition, bioactive oil extract, or formulation, or the bioactive terpene composition, bioactive oil extract, or formulation can be applied to a surface, substrate (eg. soil) or the bioactive terpene composition, bioactive oil extract, or formulation can be dispersed in the plant’s immediate vicinity or environment (eg. dispersed in the air).
  • a surface, substrate eg. soil
  • the bioactive terpene composition, bioactive oil extract, or formulation can be dispersed in the plant’s immediate vicinity or environment (eg. dispersed in the air).
  • the bioactive terpene composition, bioactive oil extract, or formulation can be applied in an environment of the biological organism so as to elicit the desired effect.
  • the bioactive terpene composition, bioactive oil extract, or formulation can be applied to soil or a plant surface, or dispersed in the air so as to produce the desired effect on an insect, mite, spider etc.
  • bioactive terpene composition, bioactive oil extract, or formulation can be applied to a plant or dispersed in an environment of the plant, so as to invoke a defence mechanism in the plant against an insect, mite, spider, bacteria, fungus, virus etc.
  • the bioactive terpene composition, bioactive oil extract, or formulation can be applied to a plant or dispersed in an environment of the plant, so as to control a property of the plant, such as eliciting plant disease resistance, enhancing plant disease resistance, promoting plant growth, increasing biomass production within the plant, or increasing plant uniformity.
  • bioactive terpene composition any suitable quantity of bioactive terpene composition, bioactive oil extract, or formulation can be used.
  • a concentration of at least one Ippm of the bioactive terpene composition, bioactive oil extract, formulation, or terpene profile is applied to the biological organism or surface/substrate/plant, more preferably about l-5ppm, including all 0. Ippm increments there between.
  • the term “substantially” generally means a change of up to 10%, including all 0.01% incremental values between 0 and 10%.
  • transitional phrase “consisting essentially of’ is used to define a composition, process or method that includes materials, steps, features, components, or elements, in addition to those literally disclosed, provided that these additional materials, steps, features, components, or elements do not materially affect the basic and novel characteristic(s) of the claimed invention.
  • the term “consisting essentially of’ occupies a middle ground between “comprising” and “consisting of’.
  • essential oil means a concentrated liquid containing aromatic and/or volatile compounds derived from the flowers, panicles, stem, or upper leaves, or a combination thereof, of one or more plants.
  • prophylactic means that the method or composition is administered in order to prevent the onset of a disease or a condition such as pest infestation.
  • prevention or “prophylactic” does not necessarily imply that the plant will never contract a disease, disorder or condition such as a pest infestation.
  • Prevention or “prophylactic” may be considered to mean reducing the likelihood of onset of a disease, disorder or condition, or preventing or otherwise reducing the risk of developing a disease, disorder or condition.
  • the inventors have been breeding new cannabis cultivars primarily with a focus on the development of phenotypes with improved pesticidal properties.
  • the approach to this has been based on the selection of candidate plants from a broad genetic background created from crosses between several locally sourced Low THC hemp cultivars and then cultivated under commercial field conditions.
  • Example 2 Essential oil cannabis extract preparation
  • Essential oil extractions are generally achieved by either steam distillation or hydrodistillation.
  • Steam distillation passes steam through a bed of plant material in a closed system. Volatile compounds are carried away in the steam and then condensed.
  • Hydrodistillation immerses the plant material in water which is then boiled and the steam (containing the volatile compounds (terpenes)) condensed and collected.
  • bioactive essential oil extracts were prepared by steam distillation of raw, frozen material through a heated water still and two condensers.
  • An L-shaped recovery head adapter was attached to the horizontal glass condenser to direct terpenes into a separation flask. The separation flask was then placed below with a glass funnel inside. A bucket was placed below the separation flask to recollect water that separated from the terpenes.
  • a reference sample was developed from a sample of essential oil (steam distillation) extracts of a mixed batch of the inventors’ cultivars.
  • a gas chromatography sample was analysed by Shimadzu QP 2020 NX GCMS to identify composition and quantifying all components that were present at levels >0.1 %, based on normalised peak area of GC-FID or GC-MS response, using analytical standards or by searching against NIST El MS library database and positive matches confirmed against standard reference materials (where available).
  • Carrier Gas Nitrogen 1.2 mE/min constant flow
  • Injection Volume 1.0 pL of 1000 ppm (50 mg / 50 mL) in HPLC Grade Methanol
  • the inventors characterised cannabis cultivars whose active compounds included relatively high levels of the monoterpenes a-Pinene, P-Pinene, P-Myrcene, D-Limonene, Trans-Ocimene and Cis-Ocimene, and had desirable allelopathic effects.
  • the inventors further developed a range of relative active concentrations of the key monoterpenes as shown in Table 7B below, but which excluded P-myrcene (which is not always a key terpene).
  • Table 7B [270] The above numerical values were obtained by calculating the maximum % margin increase (maximum margin increase) that kept the proportions of the terpenes the same. The upper bound was calculated by adding the maximum margin increase to the average blended result. The lower bound was calculated by subtracting the maximum margin increase from the average blended result.
  • Example 5 Formulation containing cannabis extract
  • This Example details the composition of an oil in water emulsion formulation containing 200 g/L of the inventors’ own hemp extracts, namely Ml 83 and Ml 68.
  • the extract is formulated in a batch process.
  • the method of formulation, including the sequence of operations is described in Table 9 below.
  • Atlox 4916 will be semi-solid at ambient temperatures and requires melting. Heat in a suitable drum oven and maintain in a completely molten state prior to addition.
  • Example 6 Insecticidal properties of cannabis extract on mosquito larvae
  • Extract Ml 68 produced the best result, but Ml 83 produced similar results to extract Ml 68 (results not shown).
  • Example 7 - Insecticidal properties of cannabis extract on fruit flies [287] A key development in the inventors’ research was the development of a high- resolution laboratory assay capable of detecting fine scale differences between the efficacy of different extracts, and in the differential response of species to an extract.
  • the assay used laboratory cultures of two Drosophila species (D. melanogaster and D. .simulan.s). Of each strain ten male and ten female flies were analysed per replicate at water diluted concentrations of 0 %, 20 %, 35 %, 40 %, 45 %, 50 % and 80 %. These ranges were determined in prior pilot testing.
  • the flies were sorted by sex, anaesthetised with CO2 and placed into 60 ml glass vials at ten individuals per replicate. An agar spoon was added to each vial in order to prevent desiccation and starvation during the bioassay. A folded 70 mm Whatman® filter paper with 100 pl of each corresponding dilution was added to each vial. The vials were maintained in a 19 °C temperature chamber with a 16/8 h day/night cycle for the duration of the bioassay. Five replicates were carried out per species and sex for a total of 280 individual flies, 40 of which were negative controls.
  • the bioassay shows that the hemp extract in high concentrations is an effective insecticide for Drosophila. It also shows that the LC50 can strongly vary depending by species and even sex. Note that this is a residual assay and the relatively high concentration of extract needed to achieve LC50 is expected as the insects do not contact the extract directly. This assay has been subsequently used to test the efficacy of extracts and mixtures of extracts. [293] The research has confirmed that all the tested extracts (M183, M168 and a 50:50 blend of extracts Ml 83 and Ml 68) demonstrate insecticidal properties and that a doseresponse relationship is observed. The extracts showed differing levels of efficacy at higher concentrations in Drosphila species (although all were efficacious), and similarly at lower concentrations in mosquito larval trials.
  • Example 8 Miticidal properties of cannabis extract
  • Two Spotted Spider Mites are a major agricultural pest of a wide range of commercial crops including hemp and cannabis.
  • the formulated extract of Example 5 (formulation containing extract M183, as shown in Table 8) was sprayed on hemp plants with spider mite infestations.
  • Spider mites with known genetic backgrounds were obtained from Bio21 and reared in a purpose-built rearing facility on dwarf bean plants until the numbers were sufficient for the experiment.
  • Infested leaves of the bean plants from the pest rearing facility were removed and pinned to cannabis/hemp plants grown in a large industrial greenhouse facility and under commercial production conditions. This allowed the pests to infest the hemp plants and hence allow efficacy testing experiments.
  • the experimental protocol consisted of four treatments (control, low concentration, medium concentration and high concentration formulated M183 extract). These concentrations were 1:5000, 1:500, 1:250. Five hemp plants were sprayed with each concentration (control was water). Prior to spraying an image was taken of leaves from each plant to estimate spider mite density. Follow up images were taken every day following spraying until Day 5 when the experiment was terminated.
  • a formulation containing extract Ml 83 (as shown in Table 8 of Example 5) was diluted with distilled water, 1:500 and 1:1000. Hemp seeds were soaked for six hours in the diluted formulated extract. After soaking the seeds were removed from the treatment and placed on a piece of filter paper on a plate, one each for each treatment. The seeds were kept moist with liquid from their respective treatment for a period of seven days, under lights. Photo records were taken from day 2 to assess the rate of germination. Table 11 shows results of second seed germination trial.
  • the graph of Figure 1 shows the results of root counts for both treatments on Day 13 of the experiment.
  • the graph of Figure 2 shows the results of root area estimates for both treatments on Day 13 of the experiment.
  • allelopathic effects between species might be more pronounced the greater the phylogenetic distance between the species. That is, that the more distant species are from one another (i.e. from an evolutionary history perspective) the greater the magnitude of any inhibitory effect.
  • the early developmental stages i.e. seed - seedling
  • the aim of this experiment was to test an essential oil cannabis extract from a strain developed by the inventors for potential allelopathic effects when applied at different life cycle stages to cannabis plants.
  • allelopathic effects monitored were biomass, insect resistance and terpene profile.
  • the main experiment considered the effect of exposure to the extract at three developmental stages: vegetative, flowering and fruiting.
  • the experiment consisted of five different treatments each representing a different developmental stage(s), and a control treatment which was never treated with extract. Treatments and number of plants in each treatment were as in Table 13 below.
  • plants were maintained following normal commercial greenhouse management procedures. Plants were supplied with a varying nutrient regime in the vegetation, flowering and fruiting phases. Plants were fed according to dryness in pots and rate of nutrient uptake.
  • the pest species used in the experiment was the Black Bean Aphid (Aphis fabae), a major economic pest which has a very broad host range with over 80 host species including potato, cabbage, cauliflower, radish, celery, capsicum, eggplant, cucumber, beets, cucurbits, chilli, and grain.
  • Black Bean Aphid Aphis fabae
  • aphids can transmit certain plant viruses.
  • Table 15 below shows the gross weight of plants in each treatment as an average weight per plant for that treatment.
  • a ratio was calculated for the proportion of flower to stem (flower ratio) by dividing the average flower weight by the average stem weight for that treatment.
  • Table 17 below shows flower yields (gms) average per plant based on a supplementary trial utilising extract Ml 68 formulated as per Table 8 of Example 5.
  • Table 18 below reports the observed pest infestation levels in treatments of the main experiment (using Ml 83 formulated extract). Figures presented are the % of infested plants per treatment.
  • This experiment is the first time that research has been focused on the potential of positive allelopathic effects of cannabis extract when applied to cannabis plants. Allelopathy research has largely examined the potential of negative allelopathic effects when an extract of one plant is applied to a phylogenetically distant plant species. The purpose of that research was to identify any potential for the development of “natural” herbicides. This experiment is also the first documented example of positive allelopathy in mature plants. Previous studies have not examined life stages beyond the seedling stage.
  • the trial consisted of two extracts (from varieties M168 and M183) and a blend of the two (i.e. 50:50 M168:M183) - a total of three cannabis extracts. These were applied to the leaves through foliar application to cuttings and in the vegetative stages of cannabis plants of the Ml 68 and Ml 83 variety.
  • a control treatment for each variety was included by treating plants with a foliar spray of water only. This resulted in a total of eight treatments (see table 19 below). Each treatment had 30 plants giving a total of 240 plants in the experiment in total.
  • the two cannabis extracts used in the experiment were an extract from the flowers of varieties M168 and M183. The methods of extraction and subsequent formulation of the cannabis extracts are as previously described. A third extract was made from an equal parts blend of the M168 and M183 extracts.
  • Foliar treatments were applied using a standard pump action mist spray - administered approximately 30cm from the test subjects in each bay twice a week during the cutting and vegetative stages.
  • Bays were individually cleaned by removing residual insects and other matter from the flood tray.
  • Insect infestation levels were recorded for aphids, spider mites, caterpillars and scale. However, caterpillar and scale numbers were so low that these results have not been included. Aphid and spider mite populations were scored for each plant against the criteria in the table below and a score from 0 to 5 was given.
  • the results support a finding that treatment during the cutting and vegetation stages with a cannabis extract provides the plant with an increased resistance to pest infestation, and mould/fungal infections. Additionally, the data show that a blend of cannabis varieties in the extract results in superior protection compared to single variety extracts.
  • the bioactive essential oil extract of the present invention may be applied to and should be effective on all plants within the Canabaceae family. Primarily, this applies to cannabis, hemp and hops varieties.
  • Pesticide residue has become a major issue for the hemp and medicinal cannabis industries. There are a number of reasons for this. Firstly, the concentration of plant chemicals through the extraction process means that any pesticide residue on the raw material is concentrated in the extract. As these extracts are used in food and medicines there has been growing concern that they may have serious impacts on human health. This has occurred in the context of a broader community concern about the longer - term impacts of chemical pesticides on bees and the wider environment.
  • Hops are closely related to cannabis and are in the same family Canabaceae. As with cannabis, hops products are made using various forms of extraction and are hence susceptible to residue from chemical insecticides, fungicides etc.
  • the female flowering hop plant (Humulus lupulus) produces cones that are also called seed cones or strobiles. Non-fertilized cones are used as a bittering and flavouring agent in beer.
  • Global demand for beers with high hop content continues to grow and craft brewers are proving to be innovative in the ways they are introducing high volumes of hops into their brews. Hops are increasingly being added later in the brewing process and many craft breweries are using the addition of green, undried, high-moisture hops to the brewing process.
  • hop cultivation encounter a range of challenges including bacterial disease, fungus and mildew, viral disease, pests and parasitic invasion. The use of chemical agricultural products to counter these challenges can result in the accumulation of chemical residue and hence the concerns over human and environmental health.
  • the inventors’ have developed a bioactive cannabis/hemp (essential) oil extract that addresses a significant need in the hemp and medicinal cannabis industry, which is for a safe, residue free pest control product.
  • No pesticide product labels currently list cannabis crops (nor are any approved by regulators) as an allowable application, and only a few are currently approved for hemp fiber (many of which are neem oil/azadarichtin formulations).
  • a pesticide made from cannabis will not have any residues (i.e. residue will be indistinguishable from existing plant compounds) on cannabis plants, particularly since preferred embodiments of the invention, increase pest resistance by producing a biological response in the subject plant. Accordingly, pest resistance is preferably achieved by way of increasing production of plant defence compounds, which are native to the subject plant and therefore do not leave any residue.
  • plant defence compounds which are native to the subject plant and therefore do not leave any residue.
  • the inventors believe that their extract will service a major gap in the market.
  • the inventors believe that their extract can be used for a wide range of pests on cannabis and other plants in the Cannabaceae family.
  • a cannabis-based bioactive terpene composition comprising the following concentrations of monoterpenes:
  • a bioactive terpene composition comprising a bioactive oil extract prepared from cannabis plants, wherein the composition comprises the following concentrations of monoterpenes:
  • bioactive oil extract based on or prepared from cannabis plants, wherein the bioactive oil extract comprises the following concentration of monoterpenes:
  • a preparation step comprising preparing a bioactive oil extract from cannabis plants.
  • a formulation comprising the bioactive terpene composition of the first or second paragraph, or the bioactive oil extract of the third or fifth paragraph, and, optionally, at least one additional ingredient.
  • a method of controlling a biological organism comprising the step of treating the biological organism with or exposing the biological organism to:
  • bioactive terpene composition or bioactive oil extract is based on or prepared from cannabis plants of the species Cannabis Sativa L.
  • bioactive terpene composition or bioactive oil extract is prepared from more than one cannabis plant cultivar, variety or chemovar, preferably two.
  • pinene terpene comprises a-pinene and P-pinene.
  • bioactive terpene composition comprises or substantially comprises the monoterpene concentrations of hemp variety M183, as seen in Tables 2 or 6B.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of hemp variety M183, as seen in Tables 2 or 6B.
  • bioactive terpene composition comprises or substantially comprises the monoterpene concentrations of hemp variety M168, as seen in Tables 2 or 6D.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of hemp variety M168, as seen in Tables 2 or 6D.
  • bioactive terpene composition bioactive oil extract, formulation, or terpene profile is produced by blending the oil extracts of hemp varieties together.
  • bioactive terpene composition comprises or substantially comprises the average % terpene concentrations of M183 and M168 as seen in Table 2.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of the average % terpene concentration of M183 and M168 as seen in Table 2.
  • Terpene Profile 1 comprises any one or more of:
  • bioactive terpene composition comprises or substantially comprises the monoterpene concentrations of hemp variety Ml 83, as seen in Table 3.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of hemp variety M183, as seen in Table 3.
  • bioactive terpene composition comprises or substantially comprises the monoterpene concentrations of hemp variety M168, as seen in Table 3.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of hemp variety M168, as seen in Table 3.
  • bioactive terpene composition comprises or substantially comprises the average % terpene concentration of M183 and M168 as seen in Table 3.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of the average % terpene concentration of M183 and M168 as seen in Table 3.
  • bioactive terpene composition comprises or substantially comprises the monoterpene concentrations of hemp variety M183, as seen in Table 4.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of hemp variety M183, as seen in Table 4.
  • bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the monoterpene concentrations of hemp variety M168, as seen in Table 4.
  • bioactive terpene composition, bioactive oil extract, formulation, or terpene profile comprises or substantially comprises the essential oil constituents of hemp variety M168, as seen in Table 4.
  • bioactive terpene composition comprises or substantially comprises the average % terpene concentration of M183 and M168 as seen in Table 4.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of the average % terpene concentration of M183 and M168 as seen in Table 4.
  • Terpene Profile 2 comprises any one or more of:
  • bioactive terpene composition comprises or substantially comprises the monoterpene concentrations of hemp variety M183, as seen in Table 5.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of hemp variety M183, as seen in Table 5.
  • bioactive terpene composition comprises or substantially comprises the monoterpene concentrations of hemp variety M168, as seen in Table 5.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of hemp variety M168, as seen in Table 5.
  • bioactive terpene composition comprises or substantially comprises the average % terpene concentration of M183 and M168 as seen in Table 5.
  • bioactive terpene composition comprises or substantially comprises the essential oil constituents of the average % terpene concentration of M183 and M168 as seen in Table 5.
  • bioactive terpene composition comprises or substantially comprises the monoterpene concentration ranges substantially as shown in Table 7B.
  • bioactive terpene composition comprises or substantially comprises the terpene concentration ranges substantially as shown in Tables 4, 5 or 7A.
  • bioactive terpene composition or formulation is in the form of a liquid, solution, gel, creme, emulsion, paste, film, powder, foam, suspension, gas, vapour or aerosol.
  • bioactive terpene composition, formulation or bioactive oil extract is formulated for use as a pest repellant, pesticide, insect repellant, insecticide, mite repellant, miticide, arachnid repellant, acaricide, anti-fungal, fungicide, anti-bacterial, bactericide, antiviral, viricide, and/or for plant allelopathy (positive or negative effect).
  • bioactive terpene composition, formulation or bioactive oil extract is capable of controlling an insect, mite, spider, or a plant.
  • bioactive terpene composition, formulation or bioactive oil extract is capable of controlling mosquitoes or mosquito larvae, flies or fly larvae, aphids, mites, spiders, cockroaches, weevils or rootworms.
  • bioactive terpene composition, formulation or bioactive oil extract is capable of controlling a fungus, bacteria or virus.
  • bioactive terpene composition, formulation or bioactive oil extract is capable of controlling one or more properties of a plant, either positively or negatively.
  • bioactive terpene composition, formulation or bioactive oil extract is capable of controlling one or more properties of plant species in the Cannabaceae family, such as cannabis, hemp or hops varieties.
  • bioactive terpene composition, formulation or bioactive oil extract is capable of: stimulating the production of plant defence compounds within a plant; triggering anti-microbial defence mechanisms within a plant; activating anti-viral defence mechanisms within a plant; activating anti-bacterial defence mechanisms within a plant; eliciting plant disease resistance; enhancing plant disease resistance; promoting plant growth; increasing biomass production within a plant; increasing plant uniformity; or, use as a herbicide.
  • bioactive terpene composition, formulation or bioactive oil extract is capable of producing an allelochemical capable of interfering with the biology of the biological organism.
  • bioactive terpene composition, formulation or bioactive oil extract is applied to a surface, substrate (eg. soil) or plant surface (eg. roots, foliage, flower head, seed) on which may be found the biological organism or which may be in close proximity to the biological organism, or the bioactive terpene composition, formulation or bioactive oil extract is dispersed in the biological organism’s immediate vicinity or environment (eg. dispersed in the air).
  • substrate eg. soil
  • plant surface eg. roots, foliage, flower head, seed
  • bioactive terpene composition, formulation or bioactive oil extract is applied to a plant or dispersed in an environment of the plant, so as to invoke a defence mechanism or allelogenic chemical in the plant against an insect, mite, spider, bacteria, fungus or virus.
  • bioactive terpene composition, formulation or bioactive oil extract is applied to a plant or dispersed in an environment of the plant, so as to control a property of the plant, such as eliciting plant disease resistance, enhancing plant disease resistance, promoting plant growth, increasing biomass production within the plant, or increasing plant uniformity.
  • plant material derived or obtained from the hemp variety comprises live plant material that can be propagated, such as cuttings or seeds.

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  • Life Sciences & Earth Sciences (AREA)
  • Environmental Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Pest Control & Pesticides (AREA)
  • Plant Pathology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Botany (AREA)
  • Developmental Biology & Embryology (AREA)
  • General Health & Medical Sciences (AREA)
  • Insects & Arthropods (AREA)
  • Agronomy & Crop Science (AREA)
  • Dentistry (AREA)
  • Physiology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Dispersion Chemistry (AREA)
  • Toxicology (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Medicines Containing Plant Substances (AREA)

Abstract

La présente invention a pour objet des procédés de production d'une réponse biologique dans des plantes de la famille Cannabaceae. Le procédé peut comprendre l'étape consistant à administrer un extrait d'huile bioactif à une plante de la famille Cannabaceae, pour ainsi produire la réponse biologique. L'extrait d'huile bioactif peut être un extrait de cannabis. L'extrait de cannabis peut comprendre les concentrations suivantes de monoterpènes : un profil de terpène 1 (environ 0,3 à 50,1 % de pinène ; environ 0,1 à 10,9 % de limonène ; et environ 0,2 à 39,0 % d'ocimène), et un profil de terpène 2 (environ 4,8 à 45,6 % de pinène ; environ 1,0 à 9,0 % de limonène ; environ 3,7 à 35,5 % d'ocimène ; et environ 1,1 à 9,9 % de β-myrcène).
PCT/AU2022/051302 2021-10-28 2022-10-28 Procédé de production d'une réponse biologique Ceased WO2023070165A1 (fr)

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Citations (3)

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Publication number Priority date Publication date Assignee Title
US20110229589A1 (en) * 2008-11-27 2011-09-22 Hanan Elraz Insect and plant disease control compositions and methods of use thereof
PL214762B1 (pl) * 2009-09-23 2013-09-30 Inst Wlokien Naturalnych I Roslin Zielarskich Kompozycja przeciwko szkodnikom, zwłaszcza pluskwiakom, zastosowanie kompozycji do zwalczania szkodników oraz sposób wytwarzania kompozycji
US20220400687A1 (en) * 2021-06-10 2022-12-22 Fractal Growth LLC Use of solutions of cannabinoids for improving cannabinoid production in cannabis plants treated therewith

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
US20110229589A1 (en) * 2008-11-27 2011-09-22 Hanan Elraz Insect and plant disease control compositions and methods of use thereof
PL214762B1 (pl) * 2009-09-23 2013-09-30 Inst Wlokien Naturalnych I Roslin Zielarskich Kompozycja przeciwko szkodnikom, zwłaszcza pluskwiakom, zastosowanie kompozycji do zwalczania szkodników oraz sposób wytwarzania kompozycji
US20220400687A1 (en) * 2021-06-10 2022-12-22 Fractal Growth LLC Use of solutions of cannabinoids for improving cannabinoid production in cannabis plants treated therewith

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