Proteins toxic to hemipteran insect species

Patent metadata

Field	Value
Publication number	US-9713334-B2
Application number	US-201615015957-A
Country	US
Kind code	B2
Filing date	Feb 4, 2016
Priority date	Apr 6, 2012
Publication date	Jul 25, 2017
Grant date	Jul 25, 2017

Abstract

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.

First claim

What is claimed is:

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.

Assignees

• Monsanto Technology Llc

Inventors

• Baum James A
• Evdokimov Artem G
• Moshiri Farhad
• Rydel Timothy J
• Sturman Eric J
• Von Rechenberg Moritz
• Vu Halong
• Wollacott Andrew M
• Zheng Meiying

Classifications

• C07K14/325 Primary

  Bacillus thuringiensis crystal peptides, i.e. delta-endotoxins · CPC title

• C12N15/8286

  for insect resistance · CPC title

• C07K14/43577

  from flies · CPC title

• A01N47/44

  Guanidine; Derivatives thereof · CPC title

• A01N37/46

  N-acyl derivatives · CPC title