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EP-4739115-A1 - BIOSTIMULANT MICROORGANISM

EP4739115A1EP 4739115 A1EP4739115 A1EP 4739115A1EP-4739115-A1

Abstract

The invention relates to a biostimulant composition comprising a Bacillus amyloliquefaciens strain having incorporated a plasmid having at least 95% sequence identity to SEQ ID NO: 1, to its use in agriculture, to a method for enhancing the growth of a plant of interest and to a method for increasing the production of lipid cyclopeptides such as surfactin, iturin, and/or fengycin by a microorganism.

Inventors

  • LECLERCQ, MAXIME
  • VANDAMME, Vincent

Assignees

  • Agricells

Dates

Publication Date
20260513
Application Date
20240708

Claims (20)

  1. 1. A biostimulant composition comprising a strain of Bacillus amyloliquefaciens having incorporated a plasmid having at least 95% identity with SEQ ID NO:1.
  2. 2. The biostimulant composition according to claim 1 wherein the Bacillus strain has at least 90% identity with SEQ ID NO:5, said identity being calculated over the entire length of said SEQ ID NO:5.
  3. 3. The biostimulant composition according to claim 1 or 2 in which the Bacillus strain has incorporated a single copy of the plasmid having at least 95% identity with SEQ ID NO:1.
  4. 4. The biostimulant composition according to any preceding claim, wherein the Bacillus strain has incorporated the plasmid having at least 95% identity with SEQ ID NO:1 and said plasmid further encoding a peptide having 100% identity with SEQ ID NO:2, SEQ ID NO:3 and/or SEQ ID NO:4.
  5. 5. The biostimulant composition according to any one of the preceding claims, being a vegetable oil comprising the strain of Bacillus amyloliquefaciens or an oil-in-water emulsion comprising the strain of Bacillus amyloliquefaciens.
  6. 6. The biostimulant composition according to any one of the preceding claims 1 to 4 being in the form of granules, said granules comprising at least 40% by weight of calcium carbonate and/or magnesium carbonate (weight of calcium and/or magnesium carbonates: weight of granules free from Bacillus amyloliquefaciens), preferably said granules having a diameter of between 0.5 mm and 2 mm.
  7. 7. The biostimulant composition according to any one of the preceding claims 1 to 4 being in the form of a powder. rehydratable dryer, further comprising a bulking agent and a surfactant.
  8. 8. The biostimulant composition according to claim 7 in which the bulking agent is chosen from a silicate, preferably talc or kaolin, and/or a polysaccharide, preferably a maltodextrin.
  9. 9. A seed of a plant of interest coated with the biostimulant composition according to any one of the preceding claims 1 to 5.
  10. 10. A method for increasing the growth of plants comprising the step of administering to said plants the biostimulant composition according to any one of the preceding claims 1 to 8, or the coated seed according to claim 9.
  11. 1 1. The method of claim 10 wherein the biostimulant composition is applied in or onto the soil before said plant is planted or the seed of said plant is sown or, advantageously, substantially at the same time as the seed of said plant is sown.
  12. 12. The method of claim 10 or 11 wherein the biostimulant composition is applied to the soil after said plant has been planted or the seed of said plant has been sown.
  13. 13. The method according to any one of claims 10 to 12 in which the plant is chosen from Poaceae, legumes, cucurbitaceae, Solanaceae, Asteraceae, cruciferous plants, beetroot, carrot, tomato, strawberry, banana, kiwi, geranium, pelargonium, chrysanthemum, apple tree, pear tree, apricot tree, peach tree and vine, preferably tomato, corn, wheat, chicory or cucumber.
  14. 14. The method according to any one of claims 10 to 13 wherein the increased growth is that of the roots, the stem and/or the weight of the plant.
  15. 15. The method according to any one of claims 10 to 14 wherein the increased growth is measured at least 40, 45, 50, 55, 60, 65 days after administration of the Bacillus strain.
  16. 16. A method for increasing the production, by a strain selected from Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus licheniformis, of lipid cyclopeptides, preferably selected from surfactin, iturin, fengycin and mixtures thereof, comprising the step of transfecting said strain with a plasmid having at least 95% identity with SEQ ID NO: 1.
  17. 17. The method of claim 16 wherein the strain comprises at least 90% identity with SEQ ID NO:5, said identity being calculated over the entire length of said SEQ ID NO:5.
  18. 18. Use of a strain selected from Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus licheniformis, having incorporated a plasmid having at least 95% identity with SEQ ID NO:1 to increase the growth of plants.
  19. 19. Use of a strain selected from Bacillus subtilis, Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus licheniformis, having incorporated a plasmid encoding SEQ ID NO:2, SEQ ID NO:3 and/or SEQ ID NO:4 to increase the growth of plants.
  20. 20. Use of a strain according to claim 18 or 19 wherein the strain has at least 90% identity with SEQ ID NO:5, said identity being calculated over the entire length of said SEQ ID NO:5.

Description

BIOSTIMULATING MICROORGANISM Technical field The present invention relates to the use of a strain of Bacillus amyloliquefaciens for biostimulation purposes, as well as to biostimulation methods. Prior art Biostimulation of the growth of a plant of interest is known. In particular, various strains of the Bacillus subtilis group are marketed for this purpose. The strains of the Bacillus amyloliquefaciens group FZB42, IT45 or QST 713 (Serenade®) are known, the first two mainly for the purpose of controlling pathogenic organisms. Similarly, the experimental strain GAI has been tested for its effectiveness in biocontrol of pathogenic organisms, but without it being commercialized (Arguelles-Arias et al., 2009). In addition, the product RhiZoBas is a strain of Bacillus amyloliquefaciens marketed for biostimulation purposes. On the other hand, patent application WO 2018/122541 describes the interest of a lipopeptide isolate for biostimulation, in particular to free itself from the metabolism of the microorganism, which risks varying according to the culture conditions, which are difficult to control. Thus, the use of better controlled extracts of a particular strain in biostimulation reflects the difficulties encountered: depending on the agronomic conditions, a strain may have a biostimulant character, or not, or even have deleterious effects on the plant of interest. Certainly, some of the conditions can be controlled or measured before any possible application of the microorganism, such as the physicochemical parameters of the field: soil type, abundance of nitrogen or other nutrients, presence or absence of pathogens in the soil. However, other parameters are more random, such as weather conditions. This will influence both the metabolism of the “biostimulant” microorganism and that of the plant of interest, so that there are many parameters which risk affecting the biostimulant properties of a microorganism, or reducing their scope. A microorganism whose biostimulant effect is more robust and less dependent on agronomic conditions therefore represents a major interest. Brief summary of the invention A first aspect of the present invention relates to a biostimulant composition comprising a strain of Bacillus amyloliquefaciens having incorporated a plasmid having at least 95% identity with SEQ ID NO:1. Preferably, this strain of Bacillus amyloliquefaciens has at least 90% identity with SEQ ID NO:5, said identity being calculated over the entire length of said SEQ ID NO:5. Advantageously, this strain of Bacillus amyloliquefaciens has incorporated a single copy of the plasmid having at least 95% identity with SEQ ID NO:1. Preferably this strain of Baciiius amyloliquefaciens having incorporated the plasmid having at least 95% identity with SEQ ID NO:1, this plasmid further encoding a peptide having 100% identity with SEQ ID NO:2, SEQ ID NO:3 and/or SEQ ID NO:4. Preferably, this biostimulant composition is a vegetable oil comprising the strain of Bacillus amyloliquefaciens or an oil-in-water emulsion comprising the strain of Bacillus amyloliquefaciens. A related aspect of the present invention relates to a seed of a plant of interest coated with this biostimulant composition. Alternatively, preferably, this biostimulant composition is in the form of granules, said granules comprising at least 40% by weight of calcium carbonate and/or magnesium carbonate (weight of calcium and/or magnesium carbonates: weight of granules free of Bacillus amyloliquefaciens), preferably said granules having a diameter of between 0.5 mm and 2 mm. Alternatively, preferably, this biostimulant composition is in the form of a rehydratable dry powder, comprising a bulking agent and a surfactant (which may be a mixture of surfactants). Another related aspect of the present invention relates to a method for increasing the growth of plants comprising the step of administering to said plants this biostimulant composition. Preferably, in this method, the biostimulant composition is applied in or onto the soil before said plant is planted or the seed of said plant is sown or, advantageously, substantially at the same time as the seed of said plant is sown. Preferably, alternatively or additionally, in this method, the biostimulant composition is applied to the soil after said plant has been planted or the seed of said plant has been sown. Preferably, in this process, the plant is chosen from Poaceae, Legumes, Cucurbitaceae, Asteraceae, Solanaceae, Rapeseed, Beetroot, Carrots, Strawberries, Bananas, Kiwi, Geranium, Pelargonium, Chrysanthemum, Apple Tree, Pear Tree, Apricot Tree, Peach Tree and Vine, preferably Tomato, Corn, Wheat, Chicory or Cucumber. Advantageously, in this method, the increased growth is that of the roots, the stem and/or the weight of the plant, preferably this increased growth being measured at least 40, 45, 50, 55, 60, 65 days after the administration of the Bacillus strain. Another aspect of the present invention relates t