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CN-121992130-A - Transgenic soybean event KA-6G2 pathway 16-7 and methods of detecting same

CN121992130ACN 121992130 ACN121992130 ACN 121992130ACN-121992130-A

Abstract

The present invention relates to transgenic soybean events Or a derived line thereof, and also relates to a method for detecting soybean plants Or a derivative strain thereof, and a detection method thereof. Derived from transgenic soybean events The nucleic acid sequence of the strain comprises SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 3, SEQ ID NO. 4, SEQ ID NO. 5, SEQ ID NO. 6, SEQ ID NO. 7, SEQ ID NO. 8, SEQ ID NO. 10, SEQ ID NO. 11 and/or SEQ ID NO. 12 or a nucleic acid sequence complementary thereto. The soybean plants of the invention Or the derivative strain thereof has better herbicide tolerance and no influence on yield, and the detection method can accurately and quickly identify whether the biological sample contains transgenic soybean event Or a DNA molecule of a derived strain thereof.

Inventors

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Assignees

  • 青岛清原种子科学有限公司

Dates

Publication Date
20260508
Application Date
20251017
Priority Date
20241106

Claims (15)

  1. 1. A nucleic acid sequence, characterized in that, the nucleic acid sequence comprises: (a)SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:10、SEQ ID NO:11 And/or SEQ ID NO 12, or (B) A nucleic acid sequence complementary to (a); Preferably, the nucleic acid sequence is derived from a transgenic soybean event Or a derived line thereof; more preferably, the nucleic acid sequence is diagnostic for a soybean event Or an amplicon present in a derived line thereof.
  2. 2. A DNA construct comprising two expression cassettes, wherein, A) The first expression cassette comprises in operable linkage a promoter P-CsVMV having a nucleic acid sequence shown as SEQ ID NO. 21, a pat gene coding region having a nucleic acid sequence shown as SEQ ID NO. 22, and a terminator T-E9 having a nucleic acid sequence shown as SEQ ID NO. 23 for terminating expression of the gene; b) The second expression cassette comprises in operative linkage a promoter P-CLSV having the nucleic acid sequence shown as SEQ ID NO. 24, a TEV protease leader sequence L-TEV having the nucleic acid sequence shown as SEQ ID NO. 25, a OsPPO-k 1 gene coding region having the nucleic acid sequence shown as SEQ ID NO. 26, and a terminator T-NOS having the nucleic acid sequence shown as SEQ ID NO. 27, which terminates expression of the gene; Preferably, the nucleic acid sequence of the DNA construct comprises SEQ ID NO. 9.
  3. 3. Diagnosing transgenic soybean events A DNA probe present, wherein the DNA probe is of sufficient length to bind the nucleic acid sequence of claim 1, the DNA probe hybridizing to the nucleic acid sequence of claim 1 and not to other nucleic acid sequences under stringent hybridization conditions.
  4. 4. A DNA primer pair consisting of a first DNA primer and a second DNA primer different from the first DNA primer, the DNA primer pair being identical to a DNA primer containing a transgenic soybean event Together with the sample for amplification reaction to generate a transgenic soybean event for diagnosis of the sample The amplicon present; preferably, the amplicon comprises the nucleic acid sequence of claim 1; More preferably, the primer pair comprises SEQ ID NO. 13 and SEQ ID NO. 14, or SEQ ID NO. 15 and SEQ ID NO. 16.
  5. 5. Event detection method for detecting transgenic soybean contained in sample Comprising at least one DNA probe according to claim 3 or a DNA primer pair according to claim 4.
  6. 6. Detecting transgenic soybean events in a sample A method of DNA presence comprising: a) Contacting a sample to be tested with at least one DNA primer pair according to claim 4; b) Performing an amplification reaction sufficient to produce a DNA amplicon, and C) Detecting the presence of the DNA amplicon in the reaction; The amplicon sequence is derived from a transgenic soybean event Preferably, the DNA amplicon comprises the nucleic acid sequence of claim 1; Or the method comprises the following steps: a) Contacting a sample to be tested with the probe of claim 3; b) Allowing said sample to be detected and said probe to hybridize under stringent hybridization conditions, and C) Detecting hybridization conditions of the sample to be detected and the probe; wherein the detection can diagnose a transgenic soybean event in the sample Is present.
  7. 7. A method of protecting soybean plants from injury caused by herbicides comprising applying a herbicide comprising an effective amount of a glutamine synthetase inhibitor-based herbicide and/or a protoporphyrinogen oxidase inhibitor-based herbicide to a field in which at least one transgenic soybean plant comprising in its genome the nucleic acid sequence of SEQ ID No. 1, SEQ ID No. 10, 1049-5292 and SEQ ID No. 2, or SEQ ID No. 3, SEQ ID No. 10, 1049-5292 and SEQ ID No. 4, or SEQ ID No. 5, SEQ ID No. 10, 1049-5292 and SEQ ID No. 6 in sequence, or said transgenic soybean plant comprising in its genome SEQ ID No. 10, said transgenic soybean plant having resistance to a glutamine synthetase inhibitor-based herbicide and/or a protoporphyrinogen oxidase inhibitor-based herbicide.
  8. 8. A method of controlling weeds in a field in which soybean plants are grown, comprising applying to the field in which at least one transgenic soybean plant is grown, an effective amount of a herbicide of the glutamine synthetase inhibitor type and/or a protoporphyrinogen oxidase inhibitor type, said transgenic soybean plant comprising in its genome in sequence the nucleic acid sequence of SEQ ID No. 1, SEQ ID No. 10, positions 1049-5292 and SEQ ID No. 2, or SEQ ID No. 3, the nucleic acid sequence of SEQ ID No. 10, positions 1049-5292 and SEQ ID No. 4, or SEQ ID No. 5, the nucleic acid sequence of SEQ ID No. 10, positions 1049-5292 and SEQ ID No. 6, or said transgenic soybean plant comprising in its genome SEQ ID No. 10, said transgenic soybean plant having resistance to a herbicide of the glutamine synthetase inhibitor type and/or a protoporphyrinogen inhibitor type.
  9. 9. A method of growing a soybean plant tolerant to a glutamine synthetase inhibitor herbicide and/or a protoporphyrinogen oxidase inhibitor herbicide, comprising planting at least one soybean seed, wherein the genome of the soybean seed comprises a nucleic acid sequence of a specific region comprising, in sequence, SEQ ID NO:1, SEQ ID NO:10, nucleic acid sequences 1049-5292 and SEQ ID NO:2, or SEQ ID NO:3, SEQ ID NO:10, nucleic acid sequences 1049-5292 and SEQ ID NO:4, or SEQ ID NO:5, SEQ ID NO:10, nucleic acid sequences 1049-5292 and SEQ ID NO:6, or the nucleic acid sequence of the specific region comprises SEQ ID NO:10; Growing the soybean seeds into soybean plants; Spraying said soybean plants with an effective dose of a glutamine synthetase inhibitor herbicide and/or a protoporphyrinogen oxidase inhibitor herbicide to yield plants having reduced plant damage as compared to other plants not having the nucleic acid sequence of said specific region.
  10. 10. A method for producing a soybean plant having tolerance to a glutamine synthetase inhibitor herbicide and/or a protoporphyrinogen oxidase inhibitor herbicide, comprising crossing a soybean plant having tolerance to a glutamine synthetase inhibitor herbicide and/or a protoporphyrinogen oxidase inhibitor herbicide with another soybean plant, wherein the soybean plant comprises a genome comprising in sequence the nucleic acid sequence of SEQ ID NO. 1, SEQ ID NO. 10, 1049-5292 and SEQ ID NO. 2, or SEQ ID NO. 3, the nucleic acid sequence of SEQ ID NO. 10, 1049-5292 and SEQ ID NO. 4, or SEQ ID NO. 5, the nucleic acid sequence of SEQ ID NO. 10, 1049-5292 and SEQ ID NO. 6, thereby producing a plurality of progeny plants, wherein the genome comprises a progeny plant comprising the nucleic acid sequence of a specific region comprising the nucleic acid sequence of SEQ ID NO. 1, SEQ ID NO. 10, 1049-5292 and SEQ ID NO. 3, or the nucleic acid sequence of SEQ ID NO. 10, 1049-5292 and SEQ ID NO. 4, or the soybean plant having tolerance to a protoporphyrinogen oxidase inhibitor herbicide, wherein the specific region comprises the nucleic acid sequence of SEQ ID NO. 1, SEQ ID NO. 10-5292 and the progeny plant; Preferably, the method comprises the step of subjecting transgenic soybean events that are tolerant to glutamine synthetase inhibitor herbicides and/or protoporphyrinogen oxidase inhibitor herbicides Sexual crossing of a first parent soybean plant with a second parent soybean plant lacking resistance to a glutamine synthetase inhibitor herbicide and/or a protoporphyrinogen oxidase inhibitor herbicide, thereby producing a plurality of progeny plants; Treating said progeny plants with a glutamine synthetase inhibitor herbicide and/or a protoporphyrinogen oxidase inhibitor herbicide; selecting said progeny plants that are tolerant to a glutamine synthetase inhibitor herbicide and/or a protoporphyrinogen oxidase inhibitor herbicide;
  11. 11. A method of improving soybean plant tolerance, the method comprising: a) Constructing the DNA construct of claim 2; b) Inserting the DNA construct into the genome of a soybean cell; c) Regenerating said soybean cells into a soybean plant, and D) Selecting a soybean plant comprising the DNA construct; Preferably, improving soybean plant tolerance comprises its resistance to an effective amount of at least one herbicide, preferably a glutamine synthetase inhibitor type herbicide and/or a protoporphyrinogen oxidase inhibitor type herbicide.
  12. 12. Generating transgenic soybean events Or a derivative thereof, characterized in that said composition is soybean oil, soybean protein, soybean flour or soybean starch.
  13. 13. Generating transgenic soybean events Or a derived strain thereof, characterized in that the agricultural product or commodity is soybean oil, soybean protein, soybean flour, soybean starch, soybean meal, soybean flakes, soybean hulls, soybean milk, soybean cheese, soybean wine, an animal feed comprising soybean, a paper comprising soybean, a cheese comprising soybean, a soybean biomass or a fuel product produced using a soybean plant and a soybean plant part.
  14. 14. A plant cell, plant part, plant, seed or inanimate plant material comprising the nucleic acid sequence of claim 1.
  15. 15. The nucleic acid sequence of claim 1, the probe of claim 3, the primer pair of claim 4, the kit of claim 5, the method of claim 6, the method of claim 10, the composition of claim 12, or the agricultural or commodity product of claim 13, wherein the transgenic soybean event Is preserved in the form of seeds in China center for type culture Collection, and the preservation number is CCTCC NO: P202427.

Description

Transgenic soybean event KA-6G2 pathway 16-7 and methods of detecting same Technical Field The present invention relates to transgenic soybean events16-7 Or a derivative thereof, and also relates to a method for detecting soybean plants16-7 Or a derivative strain thereof, and a detection method thereof. Background Soybean (Glycine max (l.) merr) is one of the world's important food crops, and there has been a five thousand year cultivation history in which seeds contain abundant plant proteins. Nowadays, biotechnology has been widely applied to soybeans to improve their agronomic traits and quality. Herbicide tolerance is an important agronomic trait in soybean production, particularly to glufosinate-ammonium and PPO inhibitor-resistant herbicides. The tolerance of soybean to glufosinate and PPO inhibitor herbicides can be obtained by expressing glufosinate resistance genes (such as PAT) and PPO inhibitor resistance genes in soybean plants by transgenic methods. In addition to the functional gene itself, the choice of regulatory elements and their sequential arrangement are critical for obtaining good transformation events and their technical effects are unexpected. Furthermore, it is known that the expression of foreign genes in plants is affected by their location in the chromosome, such as the proximity of chromatin structures (e.g., heterochromatin) or transcriptional regulatory elements (e.g., enhancers) to integration sites, etc. For this reason, it is often necessary to screen a large number of events to make it possible to identify events that can be commercialized (i.e., events in which the introduced target gene is optimally expressed). Equal genes in the same type of transgenic plant (or other organism) can exhibit wide variation in expression levels between different events, and there may also be differences in spatial or temporal expression patterns. Disclosure of Invention The object of the present invention is to provide a transgenic soybean event16-7 And methods for detecting transgenic soybean events16-7 And methods of detection. The technical scheme adopted by the invention is as follows: The present invention provides a nucleic acid sequence comprising: (a)SEQ ID NO:1、SEQ ID NO:2、SEQ ID NO:3、SEQ ID NO:4、SEQ ID NO:5、SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO:10、SEQ ID NO:11 And/or SEQ ID NO 12, or (B) A nucleic acid sequence complementary to (a). In a specific embodiment, the nucleic acid sequence is derived from a transgenic soybean event16-7 Or a derivative line thereof. In another embodiment, the nucleic acid sequence is diagnostic for a soybean event16-7 Or a derivative thereof. The invention also provides a DNA construct comprising two expression cassettes, wherein, A) The first expression cassette comprises in operable linkage a promoter P-CsVMV having a nucleic acid sequence shown as SEQ ID NO. 21, a pat gene coding region having a nucleic acid sequence shown as SEQ ID NO. 22, and a terminator T-E9 having a nucleic acid sequence shown as SEQ ID NO. 23 for terminating expression of the gene; b) The second expression cassette comprises in operative connection a promoter P-CLSV of the nucleic acid sequence shown as SEQ ID NO. 24, a TEV protease leader sequence L-TEV of the nucleic acid sequence shown as SEQ ID NO. 25, a OsPPO-k 1 gene coding region of the nucleic acid sequence shown as SEQ ID NO. 26, and a terminator T-NOS of the nucleic acid sequence shown as SEQ ID NO. 27, which terminates expression of the gene. In a specific embodiment, the nucleic acid sequence of the DNA construct comprises SEQ ID NO. 9. The invention also provides a method for diagnosing transgenic soybean events16-7, Wherein the DNA probe is of sufficient length to bind to the nucleic acid sequence, the DNA probe hybridizing to the nucleic acid sequence under stringent hybridization conditions and not hybridizing to other nucleic acid sequences. The invention also provides a DNA primer pair consisting of a first DNA primer and a second DNA primer different from the first DNA primer, the DNA primer pair being identical to a DNA primer containing a transgenic soybean event16-7 Together for an amplification reaction to produce a nucleic acid sequence for diagnosing a transgenic soybean event in the sample16-7 Present an amplicon. In a specific embodiment, the amplicon comprises the nucleic acid sequence. In another embodiment, the primer pair comprises SEQ ID NO. 13 and SEQ ID NO. 14, or SEQ ID NO. 15 and SEQ ID NO. 16. The invention also provides a method for detecting an event comprising transgenic soybean in a sample16-7 Comprising at least one of said DNA probes or said DNA primer pairs. The invention also provides a method for detecting transgenic soybean events in a sample16-7, Comprising: a) Contacting the sample to be detected with said at least one DNA primer pair; b) Performing an amplification reaction sufficient to produce a DNA amplicon, and C) Detecting the presence of the DNA amplicon in the reac