Soybean resistant to cyst nematodes

• US 10,294,489 B2
• Filed: 03/18/2014
• Issued: 05/21/2019
• Est. Priority Date: 03/15/2013
• Status: Active Grant

• Current Assignee: Board of Trustees of Southern Illinois University, Curators of the University of Missouri (University of Missouri)
• Original Assignee: Board of Trustees of Southern Illinois University, Curators of the University of Missouri (University of Missouri)
• Inventors: Meksem, Khalid, Liu, Shiming, Kandoth, Pramod Kaitheri, Mitchum, Melissa G., Lightfoot, David
• Primary Examiner(s): Ibrahim, Medina A
• Assistant Examiner(s): Zhonh, Wayne
• Application Number:
  US14/218,669
• Publication Number:
  US 20140380522A1
• Time in Patent Office: 1,890 Days
• Field of Search: None
• CPC Class Codes: C07K 14/415 from plants
• CPC Class Codes: C12N 15/8285 for nematode resistance
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

A transgenic soybean plant or parts thereof, resistant to soybean cyst nematodes, transformed to express Glyma18g02570, Glyma18g02580, or Glyma18g02590, or a variant thereof. Also provided is a method of making such a plant. Also provided is an artificial DNA construct encoding Glyma18g02570, Glyma18g02580, or Glyma18g02590, or a variant thereof.

Claims

1. A transgenic soybean plant resistant to soybean cyst nematode (SCN), or a seed, plant part, or progeny thereof, the soybean plant transformed with an artificial DNA construct comprising, as operably associated components in the 5′

to 3′

direction of transcription;

(a) a promoter that functions in a soybean plant;

(b) a transcribable nucleic acid molecule comprising

(i) a nucleotide sequence at least 99% identical to SEQ ID NO;

3, wherein said nucleotide sequence comprises a C163225G mutation, and wherein said nucleotide sequence encodes a polypeptide having soluble NSF-attachment protein (SNAP) activity;

(ii) a nucleotide sequence encoding a polypeptide at least 99% identical to SEQ ID NO;

6, wherein said polypeptide comprises a D208E mutation, and wherein said polypeptide has SNAP activity, or;

(iii) a nucleotide sequence which is the full length complement of (i) or (ii); and

(c) a transcriptional termination sequence;

wherein the promoter is heterologous to the transcribable nucleic acid molecule, and the transgenic soybean plant exhibits increased SCN resistance compared to a control transgenic soybean plant expressing SEQ ID NO;

3.

2. The transgenic soybean plant of claim 1, wherein:

the nucleotide sequence further comprises one of more mutations selected from the group consisting of A164972AGGT, C164974A, C163208A, G164965C, G164968C, and A164972AGGC;

or

the encoded polypeptide further comprises one of more mutations selected from the group consisting of D286Y, D287E, −

288V, L289I, Q203K, E285Q, D286H, and −

288A.

3. The transgenic soybean plant of claim 1, wherein the transcribable nucleic acid molecule is expressed in epidermis, vascular tissue, meristem, cambium, cortex, pith, leaf, sheath, root, flower, developing ovule or seed.

4. The transgenic soybean plant of claim 1, wherein the promoter comprises an inducible promoter or a tissue-specific promoter.

5. The transgenic soybean plant of claim 4, wherein the promoter comprises a nematode-inducible promoter.

6. The transgenic soybean plant of claim 1, wherein the promoter is selected from the group consisting of factor EF1α

gene promoter;

rice tungro bacilliform virus (RTBV) gene promoter;

cestrum yellow leaf curling virus (CmYLCV) promoter;

tCUP cryptic promoter system;

T6P-3 promoter;

S-adenosyl-L-methionine synthetase promoter;

Raspberry E4 gene promoter;

cauliflower mosaic virus 35S promoter;

figwort mosaic virus promoter;

conditional heat-shock promoter;

promoter sub-fragments of sugar beet V-type H+-ATPase subunit c isoform; and

beta-tubulin promoter.

7. The transgenic soybean plant of claim 1, wherein increased SCN resistance corresponds to a decrease in SCN susceptibility of at least about 20%compared to a control transgenic soybean plant expressing SEQ ID NO:

3.

8. The transgenic soybean plant of claim 1, wherein the nucleotide sequence further comprises a G164968T mutation.

9. The transgenic soybean plant of claim 1, wherein the nucleotide sequence further comprises a A164972AGGT mutation.

10. The transgenic soybean plant of claim 1, wherein the polypeptide further comprises a D286Y mutation.

11. The transgenic soybean plant of claim 1, wherein the polypeptide further comprises a D287E and a −

288V mutation.

12. The transgenic progeny, seed, or plant part from the transgenic soybean plant of claim 1, wherein the transgenic progeny, seed, or part comprises the transcribable nucleic acid molecule.

15. The transgenic soybean plant of claim 1, wherein the nucleotide sequence further comprises a C164974A mutation.

16. The transgenic soybean plant of claim 1, wherein the polypeptide further comprises a L289I mutation.

17. The transgenic soybean plant of claim 1, wherein the nucleotide sequence further comprises a G164968T, a A164972AGGT, and a C164974A mutation.

18. The transgenic soybean plant of claim 1, wherein the polypeptide further comprises a D286Y, a D287E, a −

288V and a L289I mutation.

13. An artificial DNA construct comprising, as operably associated components in the 5′

to 3′

direction of transcription;

(a) a promoter that functions in a soybean plant;

(b) a transcribable nucleic acid molecule comprising

(i) a nucleotide sequence at least 99% identical to SEQ ID NO;

3, wherein said nucleotide sequence comprises a C163225G mutation, and wherein said nucleotide sequence encodes a polypeptide having SNAP activity,

(ii) a nucleotide sequence encoding a polypeptide least 99% identical to SEQ ID NO;

6, wherein said polypeptide comprises a D208E mutation, and wherein said polypeptide has SNAP activity, or;

(iii) a nucleotide sequence which is the full length complement of (i) or (ii); and

(c) a transcriptional termination sequence.

14. A method of increasing soybean cyst nematode (SCN) resistance of a soybean plant comprising:

transforming a soybean plant with an artificial DNA construct according to claim 13;

wherein the transformed soybean plant exhibits increased SCN resistance compared to a control transgenic soybean plant expressing SEQ ID NO;

3.

19. The artificial DNA construct of claim 13, wherein the nucleotide sequence further comprises one of more mutations selected from the group consisting of A164972AGGT, C164974A, C163208A, G164965C, G164968C, and A164972AGGC;

or

the encoded polypeptide further comprises one of more mutations selected from the group consisting of D286Y, D287E, −

288V, L289I, Q203K, E285Q, D286H and −

288A.