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 protein or peptide that protects a plant from a pathogen.

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 protein or peptide that protects a plant from a pathogen comprises a plant immune system enhancer protein or peptide.

10. The recombinant Bacillus cereus family member of claim 9 , wherein the plant immune system enhancer protein or peptide comprises a harpin, an α-elastin, a β-elastin, a cryptogein, a flagellin protein, or a flagellin peptide.

11. The recombinant Bacillus cereus family member of claim 1 , wherein the protein or peptide that protects a plant from a pathogen has antibacterial activity, antifungal activity, or both antibacterial and antifungal activity.

12. The recombinant Bacillus cereus family member of claim 11 , wherein the protein or peptide comprises a bacteriocin, a lysozyme, a conalbumin, an albumin, or a lactoferrin.

13. The recombinant Bacillus cereus family member of claim 1 , wherein the protein that protects a plant from a pathogen comprises an enzyme.

14. The recombinant Bacillus cereus family member of claim 13 , wherein the enzyme comprises a protease or a lactonase.

15. The recombinant Bacillus cereus family member of claim 14 , wherein the protease or lactonase is specific for a bacterial signaling molecule.

16. The recombinant Bacillus cereus family member of claim 15 , wherein the bacterial signaling molecule comprises a bacterial lactone homoserine signaling molecule.

17. The recombinant Bacillus cereus family member of claim 14 , wherein the protease comprises an alkaline protease, an acid protease, or a neutral protease.

18. The recombinant Bacillus cereus family member of claim 14 , wherein the lactonase comprises 1,4-lactonase, 2-pyrone-4,6-dicarboxylate lactonase, 3-oxoadipate enol-lactonase, actinomycin lactonase, deoxylimonate A-ring-lactonase, gluconolactonase L-rhamnono-1,4-lactonase, limonin-D-ring-lactonase, steroid-lactonase, triacetate-lactonase, or xylono-1,4-lactonase.

19. The recombinant Bacillus cereus family member of claim 13 , wherein the enzyme is specific for a cellular component of a bacterium or fungus.

20. The recombinant Bacillus cereus family member of claim 19 , wherein the enzyme comprises a β-1,3-glucanase, a β-1,4-glucanase, a β-1,6-glucanase, a chitosanase, a chitinase, a lyticase, a peptidase, a proteinase, a protease, a mutanolysin, a stapholysin, or a lysozyme.

21. The recombinant Bacillus cereus family member of claim 1 , wherein the protein or peptide that protects a plant from a pathogen protects the plant from a bacterial or fungal pathogen.

22. A method for protecting a plant from a pathogen comprising:

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

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