Compositions comprising recombinant bacillus cells and another biological control agent
View Patent ↗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 exosporium-producing Bacillus cells of a Bacillus cereus family member that express a fusion protein comprising:
(i) an aminocyclopropane-1-carboxylicacid deaminase: and
(ii) a targeting sequence, exosporium protein, or exosporium protein fragment wherein the targeting sequence or exosporium protein comprises:
an amino acid sequence having at least about 43% identity with amino acids 20-35 of SEQ ID NO: 1, wherein the identity with amino acids 25-35 is at least about 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 thuringiensis, Bacillus megaterium, Bacillus mycoides isolate J, Bacillus methylotrophicus, Bacillus vallismortis, Chromobacterium subtsugae, Delftia acidovorans, Streptomyces lydicus, Streptomyces colombiensis, Streptomyces galbus K61, and Penicillium bilaii in a synergistically effective amount.
2. The composition of claim 1 , wherein the recombinant Bacillus cells are derived from Bacillus thuringiensis BT013A.
3. The composition according to claim 1 , wherein the at least one biological control agent is a Bacillus thuringiensis strain.
4. The composition according to claim 3 , wherein the Bacillus thuringiensis strain is selected from the group consisting of B. thuringiensis var. israelensis, B. thuringiensis subsp. aizawai strain ABTS-1857, B. thuringiensis subsp. kurstaki strain HD-1, B. thuringiensis subsp. tenebrionis strain NB 176, B. th. var. aegyptii, B. th. var. colmeri, B. th. var. darmstadiensis, B. th. var. dendrolimus, B. th. var. galleriae, B. th. var. japonensis, B. th. subsp. Morrisoni, B. th. var. san diego, B. th. subsp. thuringiensis strain MPPL002, B. th. var. thuringiensis, or B. th. var 7216, B. th. strain BD#32 (NRRL Accession No. B-21530), or B. th. var T36, or mutants of any thereof having all the identifying characteristics of the respective strain.
5. The composition according to claim 1 , wherein the at least one biological control agent is Penicillium bilaii ATCC22348 or mutants thereof having all the identifying characteristics of the respective strain.
6. The composition according to claim 1 , wherein the at least one biological control agent is Streptomyces lydicus WYCD108 or Streptomyces lydicus WYCE108 or mutants of any thereof having all the identifying characteristics of the respective strain.
7. The composition according to claim 1 , wherein the at least one biological control agent is Delftia acidovorans RAY209 or mutants thereof having all the identifying characteristics of the respective strain.
8. The composition according to claim 1 , wherein the at least one biological control agent is Chromobacterium subtsugae strain PRAA4-1T or mutants thereof having all of the identifying characteristics of the respective strain.
9. A seed coated with the composition according to claim 1 .
10. 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 the steps of simultaneously or sequentially applying:
a) recombinant exosporium-producing Bacillus cells of a Bacillus cereus family member that express a fusion protein comprising:
(i) an aminocyclopropane-1-carboxylicacid deaminase: and
(ii) a targeting sequence, exosporium protein, or exosporium protein fragment wherein the targeting sequence or exosporium protein comprises:
an amino acid sequence having at least about 43% identity with amino acids 20-35 of SEQ ID NO: 1, wherein the identity with amino acids 25-35 is at least about 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 thuringiensis, Bacillus megaterium, Bacillus mycoides isolate J, Bacillus methylotrophicus, Bacillus vallismortis, Chromobacterium subtsugae, Delftia acidovorans, Streptomyces lydicus, Streptomyces colombiensis, Streptomyces galbus K61, and Penicillium bilaii in a synergistically effective amount.