Proteins toxic to hemipteran insect species
The present invention discloses Hemipteran insect inhibitory proteins, methods of using such proteins, nucleotide sequences encoding such proteins, methods of detecting and isolating such proteins, and their use in agricultural systems.
1. An insect inhibitory recombinant polypeptide comprising a sequence having at least 95% amino acid identity to SEQ ID NO: 2 (TIC807), wherein said insect inhibitory recombinant polypeptide exhibits about 15 fold or greater Hemipteran inhibitory activity against a Hemipteran pest species relative to the Hemipteran inhibitory activity of SEQ ID NO: 2, and wherein said insect inhibitory recombinant polypeptide comprises:
a) at least one modification at an amino acid residue selected from the group consisting of Val10, Asn12, Ile14, Asn22, Asn23, Gly24, Ile25, Gln26, Gly27, Phe30, Gln38, Ile39, Asp40, Thr41, Ile43, Ile52, Thr93, Ser95, Ser97, Phe138, Phe147, Gln149, Ser151, Ser159, Asn180, Thr182, Ser193, Thr194, Glu195, His196, Tyr197, Ser198, His199, Tyr200, Ser201, Gly202, Tyr203, Pro204, Ile205, Leu206, Thr207, Trp208, Ile209, Ser210, Tyr216, Gly218, Pro219, Met221, Ser222, Trp223, Tyr224, Phe225, Asn239, Val244, Tyr246, Thr247, Val251, Gln253, Ser255, and His287 relative to SEQ ID NO:2; and
(b) a contiguous triple deletion in residue range 196-201 relative to SEQ ID NO:2.
2. The insect inhibitory recombinant polypeptide of claim 1 , wherein said insect inhibitory recombinant polypeptide exhibits from about 20 fold to about 260 fold increased Hemipteran inhibitory activity against a Hemipteran pest species relative to the Hemipteran inhibitory activity of SEQ ID NO: 2.
3. The insect inhibitory recombinant polypeptide of claim 2 , wherein said insect inhibitory recombinant polypeptide exhibits about 25 fold or greater Hemipteran inhibitory activity against a Hemipteran pest species relative to the Hemipteran inhibitory activity of SEQ ID NO: 2 (TIC807).
4. The insect inhibitory recombinant polypeptide of claim 2 , wherein said insect inhibitory recombinant polypeptide exhibits about 50 fold or greater Hemipteran inhibitory activity against a Hemipteran pest species relative to the Hemipteran inhibitory activity of SEQ ID NO: 2 (TIC807).
5. The insect inhibitory recombinant polypeptide of claim 1 , wherein said insect inhibitory recombinant polypeptide comprises at least one mutation in SEQ ID NO:2 selected from the group consisting of N12D, I52M, T93A, S95A, F138V, F147S, F147A, Q149E, Q149A, S151A, S159T, N180D, T182A, H196A, Y197A, S198A, W208A, P219R, W223Y, Y246F, T247K, V251A, and H287F.
6. The insect inhibitory recombinant polypeptide of claim 1 , wherein said insect inhibitory recombinant polypeptide comprises at least two mutations in SEQ ID NO:2 selected from the group consisting of T93A, S95A, F147S, F147A, Q149E, Q149A, S151A, N180D, T182A, H196A, Y197A, S198A, W208A, S217N, P219R, W223Y, V251A.
7. The insect inhibitory recombinant polypeptide of claim 1 , wherein said insect inhibitory recombinant polypeptide comprises at least three mutations in SEQ ID NO:2 selected from the group consisting of T93A, S95A, F147S, F147A, Q149E, Q149A, S151A, N180D, T182A, H196A, S198A, Y197A, W208A, S217N, P219R, W223Y, V251A.
8. The insect inhibitory recombinant polypeptide of claim 1 , wherein said insect inhibitory recombinant polypeptide comprises at least four mutations in SEQ ID NO:2 selected from the group consisting of T93A, S95A, F147S, F147A, Q149E, Q149A, S151A, N180D, T182A, H196A, Y197A, S198A, W208A, S217N, P219R, W223Y, V251A.
9. The insect inhibitory recombinant polypeptide of claim 1 , wherein said Hemipteran pest species is selected from the group consisting of a Lygus sp., an Empoasca sp., an Amrasca sp, and a combination thereof; and wherein said Hemipteran pest species is selected from the group consisting of Lygus hesperus, Lygus Lineolaris, Amrasca devastans , and a combination thereof.
10. The insect inhibitory recombinant polypeptide of claim 1 , comprising an enhanced inhibitory spectrum against Lygus, Empoasca, Amrasca , or a combination thereof.
11. The insect inhibitory recombinant polypeptide of claim 1 , wherein said insect inhibitory recombinant polypeptide exhibits an enhanced Lygus inhibitory spectrum relative to TIC807 in that said insect inhibitory recombinant polypeptide provides increased inhibition of Lygus lineolaris.