Fusion proteins and methods for stimulating plant growth, protecting plants from pathogens, and immobilizing

1. A recombinant Bacillus cereus family member that expresses a fusion protein, wherein the fusion protein comprises:

a targeting sequence or exosporium protein that is capable of targeting the fusion protein to the exosporium of the recombinant Bacillus cereus family member; and

at least one plant growth stimulating protein or peptide, wherein the plant growth stimulating protein or peptide comprises a chitosanase, a protease, a phosphatase, a nitrogenase, a nuclease, a glucosidase, a flagellin protein, or a flagellin peptide.

2. The recombinant Bacillus cereus family member of claim 1 , wherein the fusion protein comprises a targeting sequence 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%.

3. The recombinant Bacillus cereus family member of claim 1 , wherein the fusion protein comprises a targeting sequence comprising an amino acid sequence having at least 50% identity with amino acids 20-35 of SEQ ID NO: 1, wherein the identity with amino acids 25-35 is at least 63%.

4. The recombinant Bacillus cereus family member of claim 1 , wherein the fusion protein comprises a targeting sequence comprising an amino acid sequence having at least 62% identity with amino acids 20-35 of SEQ ID NO: 1, wherein the identity with amino acids 25-35 is at least 72%.

5. The recombinant Bacillus cereus family member of claim 1 , wherein the fusion protein comprises a targeting sequence comprising an amino acid sequence having at least 81% identity with amino acids 20-35 of SEQ ID NO: 1, wherein the identity with amino acids 25-35 is at least 90%.

6. The recombinant Bacillus cereus family member of claim 1 , wherein the targeting sequence consists of: (a) a 16 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%; (b) amino acids 1-35 of SEQ ID NO: 1; (c) amino acids 20-35 of SEQ ID NO: 1; or (d) SEQ ID NO: 1.

7. The recombinant Bacillus cereus family member of claim 1 , wherein the fusion protein comprises:

(a) a targeting sequence comprising amino acids 1-35 of SEQ ID NO: 1;

(b) a targeting sequence comprising amino acids 20-35 of SEQ ID NO: 1;

(c) a targeting sequence comprising SEQ ID NO: 1;

(d) an exosporium protein comprising SEQ ID NO: 2;

(e) a targeting sequence comprising amino acids 1-27 of SEQ ID NO: 3;

(f) a targeting sequence comprising amino acids 12-27 of SEQ ID NO: 3;

(g) a targeting sequence comprising SEQ ID NO: 3;

(h) an exosporium protein comprising SEQ ID NO: 4;

(i) a targeting sequence comprising amino acids 1-38 of SEQ ID NO: 5;

(j) a targeting sequence comprising amino acids 23-38 of SEQ ID NO: 5;

(k) a targeting sequence comprising SEQ ID NO: 5;

(l) an exosporium protein comprising SEQ ID NO: 6;

(m) a targeting sequence comprising amino acids 1-28 of SEQ ID NO: 7;

(n) a targeting sequence comprising amino acids 13-28 of SEQ ID NO: 7;

(o) a targeting sequence comprising SEQ ID NO: 7;

(p) an exosporium protein comprising SEQ ID NO: 8;

(q) a targeting sequence comprising amino acids 1-24 of SEQ ID NO: 9;

(r) a targeting sequence comprising amino acids 9-24 of SEQ ID NO: 9;

(s) a targeting sequence comprising SEQ ID NO: 9;

(t) an exosporium protein comprising SEQ ID NO: 10;

(u) a targeting sequence comprising amino acids 1-33 of SEQ ID NO: 11;

(v) a targeting sequence comprising amino acids 18-33 of SEQ ID NO: 11;

(w) a targeting sequence comprising SEQ ID NO: 11;

(x) an exosporium protein comprising SEQ ID NO: 12;

(y) a targeting sequence comprising amino acids 1-33 of SEQ ID NO: 13;

(z) a targeting sequence comprising amino acids 18-33 of SEQ ID NO: 13;

(aa) a targeting sequence comprising amino SEQ ID NO: 13;

(ab) an exosporium protein comprising SEQ ID NO: 14;

(ac) a targeting sequence comprising amino acids 1-43 of SEQ ID NO: 15;

(ad) a targeting sequence comprising amino acids 28-43 of SEQ ID NO: 15;

(ae) a targeting sequence comprising amino SEQ ID NO: 15;

(af) an exosporium protein comprising SEQ ID NO: 16;

(ag) a targeting sequence comprising amino acids 1-27 of SEQ ID NO: 17;

(ah) a targeting sequence comprising amino acids 12-27 of SEQ ID NO: 17;

(ai) a targeting sequence comprising amino SEQ ID NO: 17;

(aj) an exosporium protein comprising SEQ ID NO: 18;

(ak) a targeting sequence comprising amino acids 1-33 of SEQ ID NO: 19;

(al) a targeting sequence comprising amino acids 18-33 of SEQ ID NO: 19;

(am) a targeting sequence comprising amino SEQ ID NO: 19;

(an) an exosporium protein comprising SEQ ID NO: 20;

(ao) a targeting sequence comprising amino acids 1-33 of SEQ ID NO: 21;

(ap) a targeting sequence comprising amino acids 18-33 of SEQ ID NO: 21;

aq) a targeting sequence comprising amino SEQ ID NO: 21;

(ar) an exosporium protein comprising SEQ ID NO: 22;

(as) a targeting sequence comprising amino acids 1-24 of SEQ ID NO: 23;

(at) a targeting sequence comprising amino acids 9-24 of SEQ ID NO: 23;

(au) a targeting sequence comprising amino SEQ ID NO: 23;

(av) an exosporium protein comprising SEQ ID NO: 24;

(aw) a targeting sequence comprising amino acids 1-24 of SEQ ID NO: 25;

(ax) a targeting sequence comprising amino acids 9-24 of SEQ ID NO: 25;

(ay) a targeting sequence comprising amino SEQ ID NO: 25;

(az) an exosporium protein comprising SEQ ID NO: 26;

(ba) a targeting sequence comprising amino acids 1-30 of SEQ ID NO: 27;

(bb) a targeting sequence comprising amino acids 15-30 of SEQ ID NO: 27;

(bc) a targeting sequence comprising amino SEQ ID NO: 27;

(bd) an exosporium protein comprising SEQ ID NO: 28;

(be) a targeting sequence comprising amino acids 1-33 of SEQ ID NO: 29;

(bf) a targeting sequence comprising amino acids 18-33 of SEQ ID NO: 29;

(bg) a targeting sequence comprising amino SEQ ID NO: 29;

(bh) an exosporium protein comprising SEQ ID NO: 30;

(bi) a targeting sequence comprising amino acids 1-24 of SEQ ID NO: 31;

(bj) a targeting sequence comprising amino acids 9-24 of SEQ ID NO: 31;

(bk) a targeting sequence comprising amino SEQ ID NO: 31;

(bl) an exosporium protein comprising SEQ ID NO: 32;

(bm) a targeting sequence comprising amino acids 1-15 of SEQ ID NO: 33;

(bn) a targeting sequence comprising amino SEQ ID NO: 33;

(bo) an exosporium protein comprising SEQ ID NO: 34;

(bp) a targeting sequence comprising amino acids 1-16 of SEQ ID NO: 35;

(bq) a targeting sequence comprising amino SEQ ID NO: 35; or

(br) an exosporium protein comprising SEQ ID NO: 36.

8. The recombinant Bacillus cereus family member of claim 1 , wherein the targeting sequence comprises the amino acid sequence GXT at its carboxy terminus, wherein X is any amino acid.

9. The recombinant Bacillus cereus family member of claim 1 , wherein the plant growth stimulating protein or peptide comprises a protease selected from the group consisting of subtilisin, an acid protease, an alkaline protease, a proteinase, a peptidase, an endopeptidase, an exopeptidase, a thermolysin, a papain, a pepsin, a trypsin, a pronase, a carboxylase, a serine protease, a glutamic protease, an aspartate protease, a cysteine protease, a threonine protease, or a metalloprotease.

10. The recombinant Bacillus cereus family member of claim 1 , wherein the plant growth stimulating protein or peptide comprises a phosphatase selected from the group consisting of a phosphoric monoester hydrolase, a phosphomonoesterase, a phosphoric diester hydrolase, a phosphodiesterase, a triphosphoric monoester hydrolase, a phosphoryl anhydride hydrolase, a pyrophosphatase, a phytase, a trimetaphosphatase, or a triphosphatase.

11. The recombinant Bacillus cereus family member of claim 1 , wherein the plant growth stimulating protein or peptide comprises a flagellin protein or a flagellin peptide.

12. The recombinant Bacillus cereus family member of claim 1 , wherein the plant growth stimulating protein or peptide comprises a chitosanase.

13. The recombinant Bacillus cereus family member of claim 1 , wherein the recombinant Bacillus cereus family member comprises Bacillus anthracis, Bacillus cereus, Bacillus thuringiensis, Bacillus mycoides, Bacillus pseudomycoides, Bacillus weihenstephensis , or a combination thereof.

14. The recombinant Bacillus cereus family member of claim 13 , wherein the recombinant Bacillus cereus family member comprises Bacillus thuringiensis.

15. The recombinant Bacillus cereus family member of claim 1 , wherein the plant growth stimulating protein or peptide is physically attached to the exosporium of the recombinant Bacillus cereus family member.

16. The recombinant Bacillus cereus family member of claim 1 , wherein the recombinant Bacillus cereus family member is inactivated.

17. A formulation comprising the recombinant Bacillus cereus family member of claim 1 and an agriculturally acceptable carrier.

18. A method for stimulating plant growth comprising:

introducing the recombinant Bacillus cereus family member of claim 1 or a formulation comprising the recombinant Bacillus cereus family member of claim 1 and an agriculturally acceptable carrier into a plant growth medium; or

applying the recombinant Bacillus cereus family member of claim 1 or a formulation comprising the recombinant Bacillus cereus family member of claim 1 , and an agriculturally acceptable carrier to foliage of a plant, a plant seed, or an area surrounding a plant.