Compositions comprising recombinant bacillus cells and another biological control agent

Patent metadata

Field	Value
Publication number	US-9826743-B2
Application number	US-201514857176-A
Country	US
Kind code	B2
Filing date	Sep 17, 2015
Priority date	Sep 17, 2014
Publication date	Nov 28, 2017
Grant date	Nov 28, 2017

Abstract

The present invention relates to a composition comprising a) recombinant exosporium-producing Bacillus cells that express a fusion protein comprising: (i) at least one plant growth stimulating protein or peptide and (ii) a targeting sequence that localizes the fusion protein to the exosporium of the Bacillus cells; and b) at least one further biological control agent selected from particular microorganisms disclosed herein and/or a mutant of a specific strain of a microorganism disclosed herein having all identifying characteristics of the respective strain, and/or at least one metabolite produced by the respective strain, and/or at least one metabolite produced by the respective strain that exhibits activity against insects, mites, nematodes and/or phytopathogens in a synergistically effective amount. Furthermore, the present invention relates to the use of this composition as well as a method for enhancing plant growth, promoting plant health, and/or reducing overall damage of plants and plant parts.

First claim

What is claimed is:

1. A composition comprising: a) recombinant Bacillus cereus family member cells that express a fusion protein comprising: (i) at least one plant growth stimulating protein or peptide selected from the group consisting of an endoglucanase with at least 85% sequence identity to SEQ ID NO: 107 and a phospholipase with at least 85% sequence identity to SEQ ID NO: 108; and (ii) a targeting sequence, exosporium protein, or exosporium protein fragment comprising an amino acid sequence having at least 43% identity with amino acids 20-35 of SEQ ID NO: 1, wherein the identity with amino acids 25-35 is at least 54%: a targeting sequence comprising amino acids 1-35 of SEQ ID NO: 1: a targeting sequence comprising amino acids 20-35 of SEQ ID NO: 1: a targeting sequence comprising amino acids 22-31 of SEQ ID NO: 1: a targeting sequence comprising amino acids 22-33 of SEQ ID NO: 1: a targeting sequence comprising amino acids 20-31 of SEQ ID NO: 1: a targeting sequence comprising SEQ ID NO: 1: or an exosporium protein comprising an amino acid sequence having at least 85% identity with SEQ ID NO: 2; and b) at least one biological control agent selected from the group consisting of Bacillus subtilis QST713, Bacillus subtilis MBI600, Bacillus amyloliquefaciens D747, Bacillus amyloliquefaciens FZB42, Bacillus subtilis var. amyloliquefaciens FZB24, and Bacillus subtilis Y1336 in a synergistically effective amount, wherein the synergy is between a) and b).

2. The composition according to claim 1 , wherein the Bacillus cereus family member is selected from the group consisting of Bacillus anthracis, Bacillus cereus, Bacillus thuringiensis, Bacillus mycoides, Bacillus pseudomycoides, Bacillus samanii, Bacillus gaemokensis, Bacillus weihenstephensis and Bacillus toyoiensis.

3. The composition of claim 1 , wherein the fusion protein comprises SEQ ID NO: 107.

4. The composition of claim 1 , wherein the recombinant Bacillus cereus family member cells are obtained from Bacillus thuringiensis BT013A.

5. The composition of claim 1 , wherein the fusion protein comprises SEQ ID NO: 108.

6. The composition according to claim 1 , wherein the fusion protein is expressed under the control of a sporulation promoter native to the targeting sequence, exosporium protein, or exosporium protein fragment of the fusion protein.

7. The composition according to claim 6 , wherein the fusion protein is expressed under the control of a high-expression sporulation promoter.

8. The composition of claim 7 , wherein the high-expression sporulation promoter comprises a sigma-K sporulation-specific polymerase promoter sequence.

9. The composition according to claim 6 , wherein the sporulation promoter comprises a nucleic acid sequence having at least 80% identity with a nucleic acid sequence of any one of SEQ ID NOs: 85-103.

10. The composition according to claim 1 , wherein the at least one biological control agent is Bacillus subtilis QST713.

11. The composition of claim 1 wherein the at least one plant growth stimulating protein or peptide is endoglucanase with at least 90% sequence identity to SEQ ID NO: 107.

12. The composition of claim 1 wherein the at least one plant growth stimulating protein or peptide is endoglucanase with at least 95% sequence identity to SEQ ID NO: 107.

13. The composition of claim 1 wherein the at least one plant growth stimulating protein or peptide is endoglucanase with at least 98% sequence identity to SEQ ID NO: 107.

14. The composition of claim 1 wherein the at least one plant growth stimulating protein or peptide is phospholipase with at least 90% sequence identity to SEQ ID NO: 108.

15. The composition of claim 1 wherein the at least one plant growth stimulating protein or peptide is phospholipase with at least 95% sequence identity to SEQ ID NO: 108.

16. The composition of claim 1 wherein the at least one plant growth stimulating protein or peptide is phospholipase with at least 98% sequence identity to SEQ ID NO: 108.

17. The composition of claim 13 wherein the targeting sequence comprises amino acids 20-35 of SEQ ID NO: 1.

18. The composition of claim 16 wherein the targeting sequence comprises amino acids 20-35 of SEQ ID NO: 1.

19. The composition of claim 17 wherein the at least one biological control agent is Bacillus subtilis QST713.

20. The composition of claim 18 wherein the at least one biological control agent is Bacillus subtilis QST713.

21. The composition of claim 19 wherein the recombinant Bacillus cereus family member cells are obtained from Bacillus thuringiensis BT013A.

22. The composition of claim 20 wherein the recombinant Bacillus cereus family member cells are obtained from Bacillus thuringiensis BT013A.

Assignees

• Bayer Cropscience Lp

Inventors

• Curtis Damian
• Thompson Brian

Classifications

• A01N63/00 Primary

  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 (containing compounds of determined constitution A01N27/00 - A01N59/00; unicellular algae A01N65/03) · CPC title

• C07K14/00

  Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof · CPC title

• C07K14/32

  from Bacillus (G) · CPC title

• A01N37/46 Primary

  N-acyl derivatives · CPC title

• A01N37/00

  Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids (containing cyclopropane carboxylic acids or derivatives thereof, e.g. cyclopropane carboxylic acid nitriles, A01N53/00) · CPC title