WO1994016076A1 - Microorganismes endosymbiotiques produisant des proteines antimicrobiennes - Google Patents

Microorganismes endosymbiotiques produisant des proteines antimicrobiennes Download PDF

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Publication number
WO1994016076A1
WO1994016076A1 PCT/GB1994/000012 GB9400012W WO9416076A1 WO 1994016076 A1 WO1994016076 A1 WO 1994016076A1 GB 9400012 W GB9400012 W GB 9400012W WO 9416076 A1 WO9416076 A1 WO 9416076A1
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cys
seq
gly
protein
asn
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Adrian Christopher Dubock
Keith Adrian Powell
Sarah Bronwen Rees
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Syngenta Ltd
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Zeneca Ltd
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Priority to AU58204/94A priority Critical patent/AU5820494A/en
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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
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • 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
    • 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
    • C12N15/74—Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora

Definitions

  • This invention relates to endosymbiotic micro-organisms having the ability to produce plant-derived antimicrobial proteins.
  • 'antimicrobial' proteins are defined as proteins possessing at least one of the following activities: antifungal activity (which may include anti-yeast activity); antibacterial activity. Activity includes a range of antagonistic effects resulting in partial inhibition or death. 'Plant-derived' proteins are capable of being isolated from the seed or other parts of one or more plant species.
  • proteins with antimicrobial activity have been isolated from plant sources, and such proteins are often believed to take part in host defence mechanisms directed against invading or competing micro-organisms. Some of the proteins are well-characterised, and their amino acid sequence may be known. In some cases, the cDNA or gene encoding the protein has also been isolated and sequenced.
  • plants produce a wide array of antifungal compounds, either in a constitutive or an inducible manner.
  • proteins with antifungal properties have now been identified, including chitinases, beta-1,3-glucanases, ribosome-inactivating proteins, thionins, chitin-binding lectins and zeamatins. These proteins have gained considerable attention as they could potentially be used as biocontrol agents.
  • the chitinases (Schlu baum et al, 1986, Nature, 324, 363-367) and beta-1,3-glucanases have weak activities by themselves, and are only inhibitory to plant pathogens when applied in combination (Mauch et al, 1988, Plant Physiol, 88, 936-942).
  • the chitin-binding lectins can also be classified as rather weak antifungal factors (Broekaert et al, 1989, Science, 245, 1100-1102; Van Parijs et al, 1991, Planta, 183, 258-264).
  • Zeamatin is a more potent antifungal protein but its activity is strongly reduced by the presence of ions at physiological concentrations (Roberts and Selitnermikoff, 1990, G Gen Microbiol, 136, 2150-2155). Permatins are also known plant antifungal proteins (Vigers et al, 1991, Molec Plant-Microbe Interact, 4, 315-323; oloshuk et al, 1991, Plant Cell, 3, 619-628).
  • thionins Apel et al, 1990, Physiol Plant, 80, 315-321
  • ribosome-inactivating proteins Robots and Selitrennikoff, 1986, Biosci Rep, 6, 19-29; Leah et al, 1991, J Biol Chem, 266, 1564-1573
  • have antifungal activity and are known to be toxic for human cells Carrasco et al, 1981, Eur J Biochem, 116, 185-189; Vernon et al, 1985, Arch Biochem Biophys, 238, 18-29; Stirpe and Barbieri, 1986, FEBS Lett, 195, 1-8).
  • Ca-AMPl from Capsicum annuum Bm-AMPl from Briza maxima and related proteins found in other plants including Delphinium, Catapodium, Baptisia and Microsensis species (International Patent Application Number PCT/GB93/02179 filed on 22 October 1993);
  • Rs-AFPl antifungal protein 1 and Rs-AFP2 from seeds of Raphanus sativus (Terras FRG et al, 1992, J Biol Chem, 267:15301-13309) and related proteins such as Bn-AFPl and Bn-AFP2 from Brassica napus, Br-AFPl and Br-AFP2 from Brassica rapa, Sa-AFPl and Sa-AFP2 from Sinapis alba, At-AFPl from Arabidopsis thaliana, Dm-AMPl and Dm-AMP2 from Dahlia merckii, Cb-AMPl and Cb-AMP2 from Cnicus benedictus, Lc-AFP from Lathyrus cicera, Ct-AMPl and Ct-AMP2 from Clitoria ternatea (International Patent Application Publication Number WO93/05153 published 18 March 1993);
  • the proteins are useful as fungicides or antibiotics to improve the disease-resistance or disease-tolerance of crops either during the life of the plant or for post-harvest crop protection.
  • the proteins may be extracted from plant tissue or produced by expression within micro-organisms. Exposure of a plant pathogen to an antimicrobial protein may be achieved by application of the protein to plant parts using standard agricultural techniques (eg surface spraying). The proteins may also be used to combat fungal or bacterial disease by expression within plant bodies (rather than just at the surface).
  • DNA encoding the antimicrobial proteins (which may be a cDNA clone, a genomic DNA clone or DNA manufactured using a standard nucleic acid synthesiser) may be transformed into a plant, and the proteins expressed within transgenic plants.
  • Such a method should be generally applicable to a wide range of plant species and may be easier or more effective than other methods.
  • micro-organisms have the ability to enter into non-pathogenic endosymbiotic relationships with a plant host. These naturally-occurring micro-organisms, hereinafter called 'endophytes' , are capable of infecting the plant host and being harboured within the plant but create no visible manifestations of disease. Such organisms include mutualistic and commensalistic endophytic organisms. The range of endophytes also includes organisms which can exist in the vascular tissues of the plant and organisms which can exist within the intercellular spaces of the plant. A method of endophyte-enhanced protection of plants has been described in a series of patent applications by Crop Genetics International Corporation, which are discussed below and incorporated specifically herein by reference.
  • the endophyte may be unmodified, genetically modified (as discussed below) or formulated with other components to provide additional beneficial properties.
  • the endophyte may be genetically modified to produce agricultural chemicals.
  • genetic material is derived from an agricultural- chemical-producing micro-organism and combined with a suitable endophyte. Combination of genetic material is achieved by:
  • the source of DNA encoding the agricultural chemical is a suitable micro-organism.
  • suitable micro-organisms are described in Table I (page 27) of International Application Publication Number WO91/10363 and include a wide variety of micro-organisms producing antibiotics, antifungal agents, antibacterial agents, antiviral agents, insecticides, nematocides, miticides, herbicides, fertilisers (nitrogen-fixing or phosphate solubilising agents), plant growth regulators or anti-feeding agents.
  • Suitable endophytes include Agrobacterium tumefaciens, Erwinia carotovora, Pseudomonas solanacearum, Pseudomonas syringae, Xanthomonas campestris, Streptomyces ipomoea for dicotyledonous plants; Erwinia stewartii, Xanthomonas campestris, Azospirillum lipoferum, Azospirillum brasilense, Pseudomonas syringae for monocotyledonous plants.
  • Clavibacter xyli subsp. xyli and Clavibacter xyli subsp. cynodontis (Cxc) are particularly useful for grasses such as maize, sorghum and the like.
  • the agricultural-chemical-producing endophytes may be used to enhance disease protection in any plant, including those producing fruit, vegetables and flowers, trees, field and row plants such as corn, sorghum, wheat, barley, oats, rice, brome grass, sugar cane, cotton, potatoes, tomatoes, cabbage, cauliflower, broccoli, melons, cucumbers.
  • International Application Publication Number WO88/09114 (published 1 December 1988) describes plants colonised by beneficial endophytic micro-organisms obtained by germination of seeds impregnated with the endophytes.
  • the endophyte may be a strain of the genus Clavibacter or Rhizobium, and may be genetically modified to produce an agricultural chemical.
  • the seed may be from the Gramineae, Leguminosae or Halvaceae family.
  • International Application Publication Number WO91/11907 (published 22 August 1991) describes the production of modified seed (particularly rice) containing an unmodified or modified endophyte (particularly Cxc) to produce a plant of reduced stature.
  • INCIDE Technology Crop Genetics International have already developed a corn bioinsecticide based upon this endophyte technology (trademark: INCIDE Technology).
  • the INCIDE bioinsecticide consists of the endophyte Clavibacter xyli subsp. cynodonits (Cxc) which has been genetically modified with an endotoxin gene derived from the bacterium Bacillus thuringiensis, and thus expresses a protein which is toxic to certain insect larvae. If corn seed is inoculated with the INCIDE vaccine, the modified Cxc inhabits the vascular tissue of plants grown from this seed and the crop is protected from attack by cornborer larvae. However, there may be an associated yield reduction in certain crop species or varieties (Agrow, 13/11/92, no 172, p 6).
  • European Patent Application Publication Number 185005 (Monsanto Co, published 18 June 1986) also describes a "plant-colonizing micro-organism" (herein called an endophyte) which has been genetically modified to express a B thuringiensis protein.
  • the source of DNA encoding the agricultural chemical is a suitable micro-organism. Plant-derived DNA sequences encoding antimicrobial proteins have not previously been used to modify the endophytes.
  • plant-derived antimicrobial proteins may be produced within the crop plant by expression of a gene incorported into the plant genome. This may involve over-expression of an inherent protein or expression of a protein derived from another plant species.
  • a method of producing antimicrobial-protein- producing micro-organisms capable of entering into endosymbiotic relationships with a plant host comprising the combination of genetic material encoding a plant-derived antimicrobial protein with an endophyte.
  • antimicrobial- protein-producing micro-organisms produced according to the method of the invention, and seed and plants treated with said micro-organisms.
  • Antimicrobial protein may thus be expressed within the plant by an endophyte rather than being directly expressed by the host crop plant.
  • plants which may be protected using the antimicrobial-protein-producing micro-organisms include field crops, cereals, fruit and vegetables such as: canola, oil seed rape, sunflower, tobacco, sugarbeet, cotton, soya, maize, wheat, barley, rice, sorghum, tomatoes, mangoes, peaches, apples, pears, strawberries, bananas, melons, potatoes, carrot, lettuce, cabbage, onion.
  • DNA encoding any plant-derived antimicrobial protein may be used in the method according to the invention (for example, DNA encoding chitinases, hevein, lectins, thionins, etc).
  • DNA encoding the following plant-derived antimicrobial proteins may be used in the method according to the invention: Mj-AMPl, MJ-AMP2, Ac-AMPl, AC-AMP2, Ca-AMPl, Bm-AMPl, Rs-AFPl, RS-AFP2, Br-AFPl, Br-AFP2, Bn-AFPl, Bn-AFP2, Sa-AFPl, Sa-AFP2, At-AFPl, Dm-AMPl, Dm-AMP2, Cb-AMPl, Cb-AMP2, Lc-AFP, Ct-AMPl, Ct-AMP2, Rs-nsLTP.
  • These proteins show a high level and wide spectrum of antifungal activity, and will be particularly useful for improving disease-resistance or disease-tolerance in crops.
  • one or more of these potent antimicrobial proteins may be used in conjunction with a slower-growing endophyte as a relatively low dose of the highly active protein may be needed to provide disease protection.
  • the presence of a slower-growing endophyte may result in less diversion of the host plant's metabolic resources, maintaining crop yield.
  • use of these potent plant-derived antimicrobial proteins may extend the range of plant hosts most suitable as targets for this type of disease protection. Even endophytes which are relatively poor colonisers of certain plant species (such as Cxc on wheat) may be engineered to express one or more of the potent proteins to give the desired level of protection to the host plant.
  • SEQ ID NO:l is the amino acid sequence of Mj -AMPl .
  • SEQ ID NO:2 is the amino acid sequence of MJ-AMP2.
  • SEQ ID NO: 3 is the nucleotide sequence of Mj-AMPl.
  • SEQ ID NO: is the amino acid sequence of Mj-AMPl deduced from SEQ ID NO: 3.
  • SEQ ID NO: 5 is the nucleotide sequence of MJ-AMP2.
  • SEQ ID NO:6 is the amino acid sequence of MJ-AMP2 deduced from SEQ ID NO: 5.
  • SEQ ID NO:7 is the amino acid sequence of Ac-AMPl .
  • SEQ ID NO:8 is the amino acid sequence of Ac-AMP2.
  • SEQ ID NO:9 is the nucleotide sequence of Ac-AMP2.
  • SEQ ID NO: 10 is the amino acid sequence of AC-AMP2 deduced from SED ID NO:9.
  • SEQ ID NO:11 is the amino acid sequence of Ca- AMPl.
  • SEQ ID NO:12 is one possible predicted DNA sequence for the Ca-AMPl gene.
  • SEQ ID NO:13 is the amino acid sequence of Bm-AMPl .
  • SEQ ID NO:14 is one possible predicted DNA sequence for the Bm-AMPl gene.
  • SEQ ID NO:15 is the amino acid sequence of Rs-AFPl.
  • SEQ ID NO:16 is the amino acid sequence of RS-AFP2.
  • SEQ ID NO:17 is the amino acid sequence of Br-AFPl.
  • SEQ ID NO:18 is the amino acid sequence of Br-AFP2.
  • SEQ ID NO: 19 is the amino acid sequence of Bn-AFPl .
  • SEQ ID NO:20 is the amino acid sequence of Bn-AFP2.
  • SEQ ID NO:21 is the amino acid sequence of Sa-AFPl.
  • SEQ ID NO: 22 is the amino acid sequence of Sa-AFP2.
  • SEQ ID NO:23 is the amino acid sequence of At-AFPl .
  • SEQ ID NO:24 is the amino acid sequence of Dm-AMPl.
  • SEQ ID NO:25 is the amino acid sequence of Dm-AMP2.
  • SEQ ID NO:26 is the amino acid sequence of Cb-AMPl.
  • SEQ ID NO:27 is the amino acid sequence of Cb-AMP2.
  • SEQ ID NO:28 is the amino acid sequence of Lc-AFP.
  • SEQ ID NO:29 is the amino acid sequence of Ct-AMPl .
  • SEQ ID NO: 30 is the amino acid sequence of Rs-nsLTP.
  • SEQ ID NO:31 is one possible predicted DNA sequence for the Dm-AMPl gene.
  • SEQ ID NO: 32 is one possible predicted DNA sequence for the Dm-AMP2 gene.
  • SEQ ID NO: 33 is one possible predicted DNA sequence for the Cb-AMPl gene.
  • SEQ ID NO: 34 is one possible predicted DNA sequence for the Cb-AMP2 gene.
  • SEQ ID NO:35 is one possible predicted DNA sequence for the Lc-AFP gene.
  • SEQ ID NO:36 is one possible predicted DNA sequence for the Ct-AMPl gene.
  • SEQ ID NO: 37 is the full length cDNA sequence o f Rs-AFPl .
  • SEQ ID NO:38 is the amino acid sequence of Rs-AFPl deduced from SEQ ID NO:37.
  • SEQ ID NO:39 is the truncated cDNA sequence of RS-AFP2.
  • SEQ ID NO: 40 is the amino acid sequence of RS-AFP2 deduced from SEQ ID NO: 39.
  • SEQ ID NO: 41 is the full length DNA sequence of PCR assisted site directed mutagenesis of Rs-AFP2.
  • SEQ ID NO:42 is the amino acid sequence of RS-AFP2 deduced from SEQ ID NO: 41.
  • Raphanus sativus Antifungal Protein 2 (RS-AFP2) by the endophyte Clavibacter xyli subsp. cynodontis (Cxc).
  • the Rs-AFP2 protein is expressed in a system analogous to that which is known to express the Bacillus thuringiensis endotoxin.
  • An oligonucleotide sequence coding for the antifungal protein Rs-AFP2 is prepared using Cxc-compatible codon ⁇ . This oligonucleotide sequence comprises appropriate restriction sites to enable it to be exchanged with the Bacillus thuringiensis endotoxin gene sequence present in the INCIDE Cxc bacterium.
  • Southern analysis is used to check that Cxc is transformed with the RS-AFP2 gene. If the result is positive, the bacterium is cultured to determine whether it is capable of expressing Rs-AFP2 protein in vitro.
  • Western analysis and antifungal assays are carried out on the fermentation products to determine whether the protein is produced in the correctly folded form as found in the native plant. It is known that the protein loses antifungal activity when it is reduced and hence unfolded.
  • Cultures of Cxc which are capable of expressing RS-AFP2 protein are used to treat rice plants by a soil drench or seed treatment method.
  • the rice plants are challenged with rice blast, Pyricularia oryzae and assessed for increased resistance to the pathogen over non-Cxc- infected plants.
  • Rs-AFP2 is known to be active against P_ oryzae in ijri vitro tests.
  • AAAAAAAAAA AAA 433 (2) INFORMATION FOR SEQ ID NO:6:
  • SEQ ID NO:36 AACCTTTGCG AGAGAGCTTC TCTTACTTGG ACTGGAAACT GCGGAAACAC TGGACATTGC 60 GATACTCAAT GCAGAAACTG GGAGTCTGCT AAGCATGGAG CTTGCCATAA GAGAGGAAAC 120 TGGAAGTGCT TCTGCTACTT CGATTGC 147

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Abstract

Procédé de production d'un microorganisme produisant des protéines antimicrobiennes qui est capable d'entrer en relation endosymbiotique avec une plante hôte, comprenant la combinaison du matériel génétique codant une protéine antimicrobienne provenant d'une plante avec un endophyte. Des exemples de puissantes protéines antimicrobiennes provenant de plante sont présentés, ainsi qu'un procédé permettant de protéger une plante hôte des maladies selon lequel on traite la plante hôte avec le microorganisme produisant des protéines antimicrobiennes.
PCT/GB1994/000012 1993-01-08 1994-01-05 Microorganismes endosymbiotiques produisant des proteines antimicrobiennes Ceased WO1994016076A1 (fr)

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AU58204/94A AU5820494A (en) 1993-01-08 1994-01-05 Antimicrobial-protein-producing endosymbiotic microorganisms

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GB9300281.4 1993-01-08
GB939300281A GB9300281D0 (en) 1993-01-08 1993-01-08 Antimicrobial-protein-producing endosymbiotic micro-organisms

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Cited By (38)

* Cited by examiner, † Cited by third party
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WO1995018229A1 (fr) * 1993-12-24 1995-07-06 Zeneca Limited Proteines antimicrobiennes
WO1996003519A1 (fr) * 1994-07-22 1996-02-08 Demeter Biotechnologies, Ltd. Produit de recombinaison par fusion de genes de peptides lytiques d'ubiquitine, produits en derivant et leur procede d'obtention
WO1997021814A1 (fr) * 1995-12-13 1997-06-19 Zeneca Limited Proteines antifongiques
WO1998026083A1 (fr) * 1996-12-13 1998-06-18 Monsanto Company Polypeptide antifongique et procedes de lutte contre les champignons pathogenes des plantes
US5955573A (en) * 1993-06-04 1999-09-21 Demegen, Inc. Ubiquitin-lytic peptide fusion gene constructs, protein products deriving therefrom, and methods of making and using same
WO2000011196A1 (fr) * 1998-08-18 2000-03-02 Syngenta Limited Polynucleotide sequences
WO2000068405A3 (fr) * 1999-05-07 2001-02-08 Du Pont Defensines vegetales
US6306390B1 (en) * 1997-03-24 2001-10-23 Ibaraki Prefecture Root endophyte having soil disease inhibitory activity, process for preparing said root endophyte, and method for inhibiting soil disease
US6605698B1 (en) 1995-12-13 2003-08-12 Syngenta Limited Antifungal peptides and composition thereof
WO2004005329A1 (fr) 2002-07-05 2004-01-15 Centre National De La Recherche Scientifique Peptide de plante a activite anti-microbienne
US6855865B2 (en) 1999-05-07 2005-02-15 E.I. Du Pont De Nemours And Company Nucleic acids encoding plant defensins and methods of use thereof
US6911577B2 (en) 2001-06-22 2005-06-28 Pioneer Hi-Bred International, Inc. Defensin polynucleotides and methods of use
US7528232B2 (en) 2005-05-20 2009-05-05 The University Of Kentucky Research Foundation Utility of phylloplanins as antibiotics, selective fungicides and for enhancing microbial resistance in crop plants
WO2012106759A1 (fr) 2011-02-07 2012-08-16 Hexima Limited Défensines de plante modifiées utiles en tant qu'agents anti-pathogènes
US9408394B2 (en) 2014-06-26 2016-08-09 Indigo Agriculture, Inc. Endophytes, associated compositions, and methods of use thereof
US9532572B2 (en) 2013-06-26 2017-01-03 Indigo Ag, Inc. Methods of use of seed-origin endophyte populations
US9545111B2 (en) 2013-11-06 2017-01-17 The Texas A & M University System Fungal endophytes for improved crop yields and protection from pests
US9687001B2 (en) 2013-02-05 2017-06-27 University Of Saskatchewan Endophytic microbial symbionts in plant prenatal care
US10136646B2 (en) 2013-06-26 2018-11-27 Indigo Ag, Inc. Agricultural endophyte-plant compositions, and methods of use
US10212944B2 (en) 2015-05-01 2019-02-26 Indigo Agriculture, Inc. Designed complex endophyte compositions and methods for improved plant traits
US10212940B2 (en) 2015-05-01 2019-02-26 Indigo Agriculture, Inc. Isolated complex endophyte compositions and methods for improved plant traits
US10271554B2 (en) 2013-12-24 2019-04-30 Ait Austrian Institute Of Technology Gmbh Plants containing beneficial endophytes
US10306890B2 (en) 2014-06-26 2019-06-04 Ait Austrian Institute Of Technology Gmbh Plant-endophyte combinations and uses therefor
US10462990B2 (en) 2014-06-20 2019-11-05 The Flinders University Of South Australia Inoculants and methods for use thereof
US10624351B2 (en) 2016-12-01 2020-04-21 Indigo Ag, Inc. Modulated nutritional quality traits in seeds
US10640783B2 (en) 2017-03-01 2020-05-05 Indigo Ag, Inc. Endophyte compositions and methods for improvement of plant traits
US10645938B2 (en) 2017-03-01 2020-05-12 Indigo Ag, Inc. Endophyte compositions and the methods for improvement of plant traits
US10667523B2 (en) 2014-12-30 2020-06-02 Indigo Ag, Inc. Seed endophytes across cultivars and species, associated compositions, and methods of use thereof
US10750711B2 (en) 2015-06-08 2020-08-25 Indigo Ag, Inc. Streptomyces endophyte compositions and methods for improved agronomic traits in plants
US10932469B2 (en) 2013-12-24 2021-03-02 Ait Austrian Institute Of Technology Method for propagating microorganisms within plant bioreactors and stably storing microorganisms within agricultural seeds
US11186527B2 (en) 2012-06-22 2021-11-30 Ait Austrian Institute Of Technology Gmbh Method for producing plant seed containing endophytic micro-organisms
US11263707B2 (en) 2017-08-08 2022-03-01 Indigo Ag, Inc. Machine learning in agricultural planting, growing, and harvesting contexts
US11589579B2 (en) 2017-09-22 2023-02-28 Biotenzz Gesellschaft Für Biotechnologie Mbh Polymeric particles containing microorganisms
US11754553B2 (en) 2013-09-04 2023-09-12 Indigo Ag, Inc. Agricultural endophyte-plant compositions, and methods of use
US11751515B2 (en) 2015-12-21 2023-09-12 Indigo Ag, Inc. Endophyte compositions and methods for improvement of plant traits in plants of agronomic importance
US11807586B2 (en) 2016-12-23 2023-11-07 The Texas A&M University System Fungal endophytes for improved crop yields and protection from pests
US11882838B2 (en) 2017-04-27 2024-01-30 The Flinders University Of South Australia Bacterial inoculants
US12075786B2 (en) 2017-09-18 2024-09-03 Indigo Ag, Inc. Markers of plant health

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EP0256682A2 (fr) * 1986-08-04 1988-02-24 Imperial Chemical Industries Plc Traitement des plantes
WO1991010363A1 (fr) * 1990-01-16 1991-07-25 Crop Genetics International Corporation Microorganismes endosymbiotiques producteurs de produits chimiques agricoles, et leur procede de preparation
EP0474601A2 (fr) * 1990-09-07 1992-03-11 Ciba-Geigy Ag Expression de gènes codant pour des protéines de plantes relatives à la pathogenèse dans des bactéries associés aux plantes

Patent Citations (3)

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Publication number Priority date Publication date Assignee Title
EP0256682A2 (fr) * 1986-08-04 1988-02-24 Imperial Chemical Industries Plc Traitement des plantes
WO1991010363A1 (fr) * 1990-01-16 1991-07-25 Crop Genetics International Corporation Microorganismes endosymbiotiques producteurs de produits chimiques agricoles, et leur procede de preparation
EP0474601A2 (fr) * 1990-09-07 1992-03-11 Ciba-Geigy Ag Expression de gènes codant pour des protéines de plantes relatives à la pathogenèse dans des bactéries associés aux plantes

Cited By (99)

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