Soybean Plant And Seed Corresponding To Transgenic Event MON87701 And Methods For Detection Thereof

• US 20090130071A1
• Filed: 11/06/2008
• Published: 05/21/2009
• Est. Priority Date: 11/15/2007
• Status: Active Grant

• Current Assignee: Monsanto Technology LLC (Bayer AG)
• Original Assignee: Monsanto Technology LLC (Bayer AG)
• Inventors: Miklos, John A., Paradise, Mark S., Gao, Ai-Guo, Dressel Toedebusch, Andrea S., Macrae, Ted C., Perlak, Frederick J., Kolacz, Kathryn H.
• Granted Patent: US 8,049,071 B2
• Time in Patent Office: Days
• US Class Current: 424/93.210
• CPC Class Codes: C12N 15/8286 for insect resistance
• CPC Class Codes: C12Q 1/6895 for plants, fungi or algae
• CPC Class Codes: Y02A 40/146 Genetically Modified [GMO] plants, e.g. transgenic plants

Abstract

The present invention provides a transgenic soybean event MON87701, and cells, seeds, and plants comprising DNA diagnostic for the soybean event. The invention also provides compositions comprising nucleotide sequences that are diagnostic for said soybean event in a biological sample, probes and primers for use in detecting nucleotide sequences that are diagnostic for the presence of said soybean event in a biological sample, and methods for detecting the presence of said soybean event nucleotide sequences in a biological sample. The invention further provides methods of growing the seeds of such soybean event into soybean plants, and methods of breeding to produce soybean plants comprising DNA diagnostic for the soybean event.

Claims

1. A DNA molecule comprising a nucleotide sequence of SEQ ID NO:

1, SEQ ID NO;

2, or complement thereof.

2. The DNA molecule of claim 1, comprising a nucleotide sequence of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

6 or complement thereof.

3. The DNA molecule of claim 2, consisting essentially of the nucleotide sequence of SEQ ID NO:

3 from positions 1 to 5757, the nucleotide sequence of SEQ ID NO;

5 from positions 1 to 6426, and the nucleotide sequence of SEQ ID NO;

4 from positions 379 to 2611, or complement thereof.

4. The DNA molecule of claim 2, consisting essentially of the nucleotide sequence of SEQ ID NO:

6 or complement thereof.

5. A soybean plant, or parts thereof, comprising the DNA molecule of any one of claims 1 to 4.

6. Seed of the soybean plant of claim 5, wherein said seed comprises said DNA molecule.

7. A composition derived from the soybean plant, or parts thereof, as set forth in claim 5, wherein said composition comprises a detectable amount of said DNA molecule, and wherein said composition is a commodity product selected from the group consisting of soybean meal, soy flour, soy protein concentrate, soy protein isolates, texturized soy protein concentrate, hydrolyzed soy protein, soybean oil and whipped topping.

8. A method of producing an insect resistant soybean plant comprising:

(a) crossing the soybean plant of claim 5 with another soybean plant;

(b) obtaining at least one progeny plant derived from the cross of (a); and

(c) selecting progeny that comprises nucleotide sequences of SEQ ID NO;

1 and SEQ ID NO;

2, wherein said selected progeny is an insect resistant soybean plant.

9. The method of claim 8, wherein said selecting step (c) includes subjecting said at least one progeny plant obtained from (b) to a nucleic acid amplification reaction, wherein progeny that produces an amplicon comprising at least one nucleotide sequence of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

5 or SEQ ID NO;

6 is selected, or subjecting said at least one progeny plant obtained from (b) to a nucleic acid hybridization reaction, wherein progeny hybridizing to a probe that hybridizes under stringent conditions with one or more DNA sequence selected from SEQ ID NO;

1 and SEQ ID NO;

2 is selected.

10. A method for protecting a soybean plant from insect infestation, comprising providing in the diet of a Lepidopteran pest of soybean an insecticidally effective amount of cell(s) or tissue(s) of the soybean plant, or parts thereof, of claim 5.

11. The method of claim 10, wherein said Lepidopteran pest is selected from the group consisting of Anticarsia, Pseudoplusia, Epinotia, Spilosoma, Helicoverpa, Spodoptera and Rachiplusia.

12. A pair of DNA molecules comprising:

a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of sufficient length of contiguous nucleotides of SEQ ID NO;

3 or SEQ ID NO;

5 or its complement to function as DNA primers or probes diagnostic for DNA extracted from soybean plant MON87701 or progeny thereof.

15. The pair of DNA molecules of any one of claims 12 to 14, wherein said first DNA molecule comprises 11 or more contiguous nucleotides of any portion of the transgene region of SEQ ID NO:

3 or SEQ ID NO;

5, or complement thereof, and said second DNA molecule comprises a similar length of a 5′

flanking soybean genomic DNA region of SEQ ID NO;

3, or complement thereof.

16. The pair of DNA molecules of claim 15, wherein said first DNA molecule comprises SEQ ID NO:

9 and said second DNA molecule comprises SEQ ID NO;

10.

17. The pair of DNA molecules of any one of claims 12 to 14, wherein said first DNA molecule comprises 11 or more contiguous nucleotides of any portion of the transgene region of SEQ ID NO:

4 or SEQ ID NO;

5, or complement thereof, and said second DNA molecule comprises a similar length of a 3′

flanking soybean genomic DNA region of SEQ ID NO;

4, or complement thereof.

18. The pair of DNA molecules of claim 17, wherein said first DNA molecule comprises SEQ ID NO:

12 and said second DNA molecule comprises SEQ ID NO;

13.

13. A pair of DNA molecules comprising:

a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of sufficient length of contiguous nucleotides of SEQ ID NO;

4 or SEQ ID NO;

5 or its complement to function as DNA primers or probes diagnostic for DNA extracted from soybean plant MON87701 or progeny thereof.

14. A pair of DNA molecules comprising:

a first DNA molecule and a second DNA molecule, wherein the DNA molecules are of sufficient length of contiguous nucleotides of SEQ ID NO;

6 or its complement to function as DNA primers or probes diagnostic for DNA extracted from soybean plant MON87701 or progeny thereof.

19. A method of detecting the presence of a DNA molecule selected from the group consisting of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

5 and SEQ ID NO;

6 in a biological sample, the method comprising;

(a) contacting said biological sample with a DNA primer pair comprising DNA primer molecules of sufficient length of contiguous nucleotides of SEQ ID NO;

3 or its complement, SEQ ID NO;

4 or its complement, SEQ ID NO;

5 or its complement, or SEQ ID NO;

6 or its complement, to function as DNA primers or probes diagnostic for DNA extracted from soybean plant MON87701 or progeny thereof;

(b) providing a nucleic acid amplification reaction condition;

(c) performing said nucleic acid amplification reaction, thereby producing a DNA amplicon molecule; and

(d) detecting said DNA amplicon molecule, wherein detection of an amplicon comprising at least one of SEQ ID NO;

1, SEQ ID NO;

2 and complement thereof is indicative of the presence of said DNA molecule in said biological sample.

20. The method of claim 19, wherein said biological sample is a DNA sample extracted from a soybean plant.

26. The method of claim 19 or 21, wherein said biological sample is selected from the group consisting of soybean meal, soy flour, soy protein concentrate, soy protein isolates, texturized soy protein concentrate, hydrolyzed soy protein and whipped topping.

21. A method of detecting the presence of a DNA molecule selected from the group consisting of SEQ ID NO:

3, SEQ ID NO;

4, SEQ ID NO;

5 and SEQ ID NO;

6 in a biological sample, the method comprising;

(a) contacting said biological sample with a DNA probe that hybridizes under stringent conditions with said DNA molecule, and does not hybridize under the stringent conditions with a biological sample not containing said DNA molecule;

(b) subjecting said biological sample and DNA probe to stringent hybridization conditions; and

(c) detecting hybridization of said DNA probe to said biological sample, wherein detection of hybridization is indicative of the presence of said DNA molecule in said biological sample.

22. The method of claim 21, wherein said biological sample is a DNA sample extracted from a soybean plant.

23. The method of claim 21 or 22, wherein said DNA probe comprises SEQ ID NO:

1 or SEQ ID NO;

2 or complement thereof.

24. The method of claim 21 or 22, wherein said DNA probe comprises SEQ ID NO:

11 or SEQ ID NO;

15.

25. The method of claims 21 to 24, wherein said DNA probe is labeled with at least one fluorophore.

27. A DNA detection kit comprising:

at least one DNA molecule of sufficient length of contiguous nucleotides homologous or complementary to SEQ ID NO;

3, SEQ ID NO;

4, SEQ ID NO;

5, or SEQ ID NO;

6 to function as a DNA primer or probe specific for soybean event MON87701 and/or its progeny.

28. The DNA detection kit of claim 27, wherein said at least one DNA molecule comprises SEQ ID NO:

1, SEQ ID NO;

2, or complement thereof.

29. The DNA detection kit of claim 28, wherein said at least one DNA molecule is SEQ ID NO:

1, SEQ ID NO;

2, or complement thereof.

30. A method of determining zygosity of DNA of a soybean plant genome comprising soybean event MON87701 in a soybean sample comprising:

(a) contacting said sample with a first primer pair of SEQ ID NO;

12 and SEQ ID NO;

13, that when used together in a nucleic acid amplification reaction with soybean event MON87701 DNA, produces an amplicon that is diagnostic for soybean event MON87701;

(b) performing a nucleic acid amplification reaction;

(c) detecting a first amplicon produced;

(d) contacting said sample with a second primer pair of SEQ ID NO;

13 and SEQ ID NO;

14, that when used together in a nucleic acid amplification reaction with soybean genomic DNA other than soybean event MON87701 DNA, produces an amplicon that is diagnostic for soybean genomic DNA other than soybean event MON87701 DNA;

(e) performing a nucleic acid amplification reaction; and

(f) detecting a second amplicon produced;

wherein detection of both the amplicon that is diagnostic for soybean event MON87701 and the amplicon that is diagnostic for soybean genomic DNA other than soybean event MON87701 DNA indicates that said sample is heterozygous for soybean event MON87701 DNA.

31. The method of claim 30, wherein said first primer pair is further used together with probe of SEQ ID NO:

15, and/or wherein said second primer pair is further used with probe of SEQ ID NO;

16.