WO2013052536A4 - Exploiting host molecules to augment the virulence of mycoinsecticides - Google Patents
Exploiting host molecules to augment the virulence of mycoinsecticides Download PDFInfo
- Publication number
- WO2013052536A4 WO2013052536A4 PCT/US2012/058543 US2012058543W WO2013052536A4 WO 2013052536 A4 WO2013052536 A4 WO 2013052536A4 US 2012058543 W US2012058543 W US 2012058543W WO 2013052536 A4 WO2013052536 A4 WO 2013052536A4
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- entomopathogenic fungus
- genetically modified
- fungus
- insect
- modified strain
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Ceased
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Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
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- 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
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- 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
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/50—Isolated enzymes; Isolated proteins
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43563—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects
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- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Biomedical Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Environmental Sciences (AREA)
- Mycology (AREA)
- Virology (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Physics & Mathematics (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Insects & Arthropods (AREA)
- Medicinal Chemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present disclosure provides methods of insect control in which fungal expression of insect-derived molecules is exploited for target-specific augmentation of entomopathogenic fungi virulence. The major advantages that depends upon the host molecule (peptide) chosen is the increase in virulence that can be tailored to be host specific, and the development of resistance that is be minimized since the host peptides and/or hormones regulate developmental processes that are species- and tissue-specific. Any adaptive mutations that arise in the targeted insect that could compensate for the fungal-expressed product during infection will lead to developmental defects or a fitness cost far greater than developing resistance to a pesticide. The methods, fungal strains, and nucleic acid constructs of the disclosure have been shown to be effective against a variety of insect host species.
Claims
1. A genetically modified strain of an entomopathogenic fungus comprising a heterologous recombinant nucleotide sequence encoding a peptide, polypeptide, or protein of a target insect host wherein, when said nucleotide sequence is expressed by the genetically modified entomopathogenic fungus when said fungus has infected the target insect host, the peptide, polypeptide, or protein increases the virulence of the fungus in the target insect compared to the virulence of a non-genetically modified strain of the entomopathogenic fungus in the target insect host.
2. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the heterologous recombinant nucleotide sequence is operably linked to a gene expression controlling region, wherein the gene expression controlling region directs production of a transcript from the heterologous recombinant nucleotide sequence in a recipient entomopathogenic fungus.
3. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the entomopathogenic fungus is of the class Hyphomycetes.
4. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the entomopathogenic fungus is virulent against the fire ant {Solenopsis invicta), a mosquito species, a Lepidopteran species, a Dipteran, or a Hemipteran species.
5. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the entomopathogenic fungus is virulent against the fire ant (Solenopsis invicta).
6. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the entomopathogenic fungus is virulent against a mosquito species.
7. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the entomopathogenic fungus is a strain of Metarhizium anisopliae or Beauveria bassiana.
8. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the heterologous recombinant nucleotide sequence encodes an amino acid sequence specifically inducing a biochemical or physiological reaction in a target insect, wherein the amino acid sequence is an insect neurohormone, an insect diuretic hormone, or a trypsin modulating oostatic factor.
9. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the heterologous recombinant nucleotide sequence encodes an insect neurohormone, or a bioactive fragment thereof.
10. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the insect neurohormone is a PBAN/pyrokinin or a bioactive fragment thereof.
11. The genetically modified strain of an entomopathogenic fungus of claim 9, wherein the heterologous recombinant nucleotide sequence encodes an insect neurohormone, or a bioactive fragment thereof, specifically inducing a biochemical or physiological reaction in the fire ant Solenopsis invicta.
12. The genetically modified strain of an entomopathogenic fungus of claim 11 , wherein the insect neurohormone is β-neuropeptide.
13. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the heterologous recombinant nucleotide sequence encodes an insect diuretic hormone.
14. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the heterologous recombinant nucleotide sequence encodes Manduca sexta diuretic hormone (MSDH).
15. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the heterologous recombinant nucleotide sequence encodes a trypsin modulating oostatic factor.
16. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the trypsin modulating oostatic factor is a trypsin modulating oostatic factor of Aedes aegyti or S. bullata.
17. The genetically modified strain of an entomopathogenic fungus of claim 1 , wherein the heterologous recombinant nucleotide sequence encodes a polypeptide having about 90% sequence similarity with a sequence selected from the group consisting of: SEQ ID Nos. 1 , 22, 23, and 24.
18. The genetically modified strain of an entomopathogenic fungus of claim 17, wherein the heterologous recombinant nucleotide sequence encodes a polypeptide having about 95% sequence similarity with a sequence selected from the group consisting of: SEQ ID Nos. 1 , 22, 23, and 24.
19. The genetically modified strain of an entomopathogenic fungus of claim 16, wherein the heterologous recombinant nucleotide sequence encodes a polypeptide having an amino acid sequence selected from the group consisting of: SEQ ID Nos. 1 , 22, 23, and 24.
20. The genetically modified strain of an entomopathogenic fungus of claim 2, wherein the gene controlling region has about 90% sequence similarity with a sequence encoding a β. bassiana chitinase gene signal peptide.
21. The genetically modified strain of an entomopathogenic fungus of claim 20, wherein the B. bassiana chitinase gene signal peptide has the amino acid sequence according to SEQ ID No.: 2.
22. An expression vector comprising a fungal gene expression controlling region nucleotide sequence operably linked to a nucleic acid encoding an insect-specific polypeptide wherein the gene expression controlling region directs production of a transcript, wherein when said transcript is expressed by a entomopathogenic fungus when said fungus has infected a target insect host, the insect-specific polypeptide increases the virulence of the fungus in the target insect compared to the virulence of a strain of the entomopathogenic fungus not expressing the insect specific polypeptide.
23. The expression vector of claim 22, wherein the gene controlling region has about 90% sequence similarity with a sequence encoding a B. bassiana chitinase gene signal peptide.
24. The expression vector of claim 23, wherein the B. bassiana chitinase gene signal peptide has the amino acid sequence according to SEQ ID No.: 2.
25. The expression vector of claim 22, wherein the insect-specific polypeptide is selected from the group consisting of: an insect neurohormone, an insect diuretic hormone, and a trypsin modulating oostatic factor, or a bioactive homolog or fragment thereof.
26. The expression vector of claim 25, wherein the insect-specific polypeptide is selected from the group consisting of: β-neuropeptide specific to the fire ant Solenopsis invicta, Manduca sexta diuretic hormone (MSDH), and a trypsin modulating oostatic factor of Aedes aegyti or S. bullata.
27. A method of increasing the virulence of an entomopathogenic fungus in a target insect host, comprising the steps of:
(a) obtaining a genetically modified strain of a entomopathogenic fungus according to claims 1 ; and
(b) delivering the genetically modified strain of entomopathogenic fungus, or spores thereof, to a target insect host desired to be infected by the fungus, wherein the fungus delivers a target insect-specific polypeptide or peptide to the target insect host, thereby increasing the virulence of the entomopathogenic fungus in the target insect host.
28. The method of claim 27, wherein the wherein the entomopathogenic fungus is Metarhizium anisopliae or Beauveria bassiana.
29. The method of claim 27, wherein the target insect host is the fire ant (Solenopsis invicta), a mosquito species, a Lepidopteran species, a Dipteran, or a Hemipteran species.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201161543345P | 2011-10-05 | 2011-10-05 | |
| US61/543,345 | 2011-10-05 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| WO2013052536A2 WO2013052536A2 (en) | 2013-04-11 |
| WO2013052536A3 WO2013052536A3 (en) | 2013-06-06 |
| WO2013052536A4 true WO2013052536A4 (en) | 2013-07-25 |
Family
ID=48044380
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/US2012/058543 Ceased WO2013052536A2 (en) | 2011-10-05 | 2012-10-03 | Exploiting host molecules to augment the virulence of mycoinsecticides |
Country Status (1)
| Country | Link |
|---|---|
| WO (1) | WO2013052536A2 (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2018140496A1 (en) * | 2017-01-24 | 2018-08-02 | Flagship Pioneering, Inc. | Methods and related compositions for manufacturing food and feed |
| CN114480300B (en) * | 2021-06-28 | 2024-12-06 | 吉林省农业科学院 | A fungal virus for improving the toxicity of Beauveria bassiana and a method for transmitting the virus |
| CN115991759A (en) * | 2022-10-20 | 2023-04-21 | 曲靖师范学院 | Paraphalocrocis medinalis sex pheromone binding protein gene GmelPBPs and application thereof |
| CN115505538B (en) * | 2022-11-17 | 2023-08-04 | 中国农业科学院植物保护研究所 | Metarhizium anisopliae strain CIPPMA0941, application thereof in preventing and treating solenopsis invicta and microbial inoculum |
| ES2978576B2 (en) * | 2023-02-09 | 2025-03-06 | Instituto Valenciano De Investig Agrarias Ivia | Machine for inoculating entomopathogenic fungal conidia in powder form onto sterile males of Ceratitis capitata |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5683689A (en) * | 1991-04-18 | 1997-11-04 | University Of Florida Research Foundation, Inc. | Controlling cockroaches, carpenter ants, and pharaoh ants using strains of beauveria bassiana |
| US5516513A (en) * | 1994-07-14 | 1996-05-14 | Troy Biosciences, Inc. | Biological ovicide for control of lepidopterous insects |
| WO1999013727A1 (en) * | 1997-09-17 | 1999-03-25 | University Of Florida | Methods and materials for control of termites |
-
2012
- 2012-10-03 WO PCT/US2012/058543 patent/WO2013052536A2/en not_active Ceased
Also Published As
| Publication number | Publication date |
|---|---|
| WO2013052536A2 (en) | 2013-04-11 |
| WO2013052536A3 (en) | 2013-06-06 |
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