Compositions comprising recombinant
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.
1. A composition comprising:
a) recombinant cells of a Bacillus cereus family member, wherein the cells express a fusion protein comprising:
(i) at least one plant growth stimulating protein or peptide selected from the group consisting of a polypeptide having endoglucanase activity and at least 85% sequence identity to SEQ ID NO: 107 and a polypeptide having phospholipase activity and at least 85% sequence identity to SEQ ID NO: 108; and
(ii) a targeting sequence, exosporium protein, or exosporium protein fragment that is capable of targeting the fusion protein to the exosporium of the recombinant Bacillus cereus family member, 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%;
amino acids 1-35 of SEQ ID NO: 1;
amino acids 20-35 of SEQ ID NO: 1;
amino acids 22-31 of SEQ ID NO: 1;
amino acids 22-33 of SEQ ID NO: 1;
amino acids 20-31 of SEQ ID NO: 1;
the amino acid sequence of SEQ ID NO: 1; or
an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 2; and
b) a Bacillus firmus strain
in a synergistic weight ratio in the range of 1:1000 to 1000:1, wherein the synergism refers to the effect of enhancing plant growth and/or promoting plant health.
2. The composition according of 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 samanil, Bacillus gaemokensis, Bacillus weihenstephensis, Bacillus toyoiensis and combinations thereof.
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 cells are 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 1 , wherein the Bacillus firmus strain is Bacillus firmus strain 1-1582.
8. A method of treating a plant, a plant part, or the locus surrounding the plant to enhance plant growth and/or promote plant health comprising applying the composition according to claim 1 .
9. 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.
10. 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.
11. 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.
12. 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.
13. 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.
14. 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.
15. The composition of claim 11 wherein the targeting sequence comprises amino acids 20-35 of SEQ ID NO:1.
16. The composition of claim 14 wherein the targeting sequence comprises amino acids 20-35 of SEQ ID NO:1.
17. The composition of claim 15 wherein the Bacillus firmus strain is Bacillus firmus strain 1-1582.
18. The composition of claim 16 wherein the Bacillus firmus strain is Bacillus firmus strain 1-1582.