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Application of herbicide tolerance protein

A herbicide and protein technology, applied in the field of degrading pyrazosulfuron-methyl herbicide by thifensulfuron-methyl hydrolase

Active Publication Date: 2016-07-06
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Only thifensulfuron-methyl hydrolase has been identified to degrade thifensulfuron-methyl, but like thifensulfuron-methyl, pyrazosulfuron-methyl also belongs to the sulfonylurea herbicides containing ester bonds, and thifensulfuron-methyl hydrolase has not been found to degrade pyrazosulfuron-methyl. Reports of tolerance to sulfuron-methyl herbicide

Method used

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  • Application of herbicide tolerance protein
  • Application of herbicide tolerance protein
  • Application of herbicide tolerance protein

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0149] The first embodiment, the acquisition and synthesis of ALT gene sequence

[0150] 1. Obtain the ALT gene sequence

[0151] The amino acid sequence (398 amino acids) of thifensulfuron hydrolase-1 (ALT-1), as shown in SEQ ID NO: 1 in the sequence listing; ALT-1-01 nucleoside encoding the amino acid sequence corresponding to said ALT-1 Acid sequence (1197 nucleotides), as shown in SEQIDNO: 2 in the sequence listing, encoding is corresponding to the ALT-1-02 nucleotide sequence (1197 nucleotides) of the amino acid sequence of said ALT-1, such as Shown in SEQ ID NO:3 in the sequence listing.

[0152] The amino acid sequence (369 amino acids) of thifensulfuron hydrolase-2 (ALT-2), as shown in SEQ ID NO: 4 in the sequence listing; ALT-2-01 nucleoside encoding the amino acid sequence corresponding to said ALT-2 Acid sequence (1110 nucleotides), as shown in SEQIDNO:5 in the sequence listing, encodes the ALT-2-02 nucleotide sequence (1110 nucleotides) corresponding to the amino...

no. 2 example

[0158] The second embodiment, construction of Arabidopsis recombinant expression vector

[0159] 1. Construction of Arabidopsis and soybean recombinant cloning vectors containing ALT nucleotide sequences

[0160] The synthetic ALT-1-01 nucleotide sequence was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN01 -T, its construction process is as follows figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ initiation codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; ALT-1-01 is the ALT-1-01 nucleotide sequence (SEQ ID NO: 2); MCS is the multiple cloning site).

[0161] Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 competent cells (...

no. 3 example

[0174] The third embodiment, the acquisition of Arabidopsis plants transferred to the ALT nucleotide sequence

[0175] 1. Transformation of recombinant expression vector into Agrobacterium

[0176] Transform the correctly constructed recombinant expression vectors DBN100632, DBN100631, DBN100634, DBN100633, DBN100636 and DBN100635 into Agrobacterium GV3101 with liquid nitrogen method, and the transformation conditions are: 100 μL Agrobacterium GV3101, 3 μL plasmid DNA (recombinant expression vector); Place in liquid nitrogen for 10 minutes, and bathe in warm water at 37°C for 10 minutes; inoculate the transformed Agrobacterium GV3101 in LB test tubes and cultivate for 2 hours at a temperature of 28°C and a rotation speed of 200rpm, and apply it to 50mg / L rifampicin Ping (Rifampicin) and 50mg / L spectinomycin LB plates until a positive single clone grows, pick the single clone culture and extract its plasmid, and carry out enzyme digestion verification with restriction endonucle...

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Abstract

The invention relate to application of herbicide tolerance protein. A weed control method includes the step of applying herbicide with an effective dosage of pyrazosulfuron ethyl to the plant growth environment of at least one transgenic plant, wherein genomes of the transgenic plants have nucleotide sequences for encoding thifensulfuron methyl hydrolase, and the transgenic plants have weak plant damage and / or increased plant yield compared with other plants without nucleotide sequences for encoding thifensulfuron methyl hydrolase. Thifensulfuron methyl hydrolase is disclosed for the first time and presents high tolerance for the pyrazosulfuron ethyl herbicide, and the plants with nucleotide sequences for encoding thifensulfuron methyl hydrolase have strong tolerance for the pyrazosulfuron ethyl herbicide and at least can bear 1-fold field concentration, and therefore wide application prospects are achieved for plants.

Description

technical field [0001] The invention relates to the use of a herbicide tolerance protein, in particular to the use of a thifensulfuron-methyl hydrolase to degrade pyrazosulfuron-methyl herbicide. Background technique [0002] Weeds can quickly deplete the soil of valuable nutrients needed by crops and other plants of interest. There are many types of herbicides currently used to control weeds, one particularly popular herbicide is glyphosate. Glyphosate-resistant crops have been developed, such as corn, soybean, cotton, sugar beet, wheat and rice. Fields of glyphosate-resistant crops can thus be sprayed with glyphosate to control weeds without significantly damaging the crop. [0003] Glyphosate has been widely used globally for more than 20 years, resulting in an overreliance on glyphosate and glyphosate-tolerant crop technologies, and in wild weed species that are naturally more tolerant to glyphosate or have developed Plants exhibiting glyphosate resistance exerted hig...

Claims

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Application Information

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IPC IPC(8): A01G13/00C12N15/82A01H5/00A01G1/00
CPCA01G2/00A01G13/00C07K14/415C07K14/80C12N9/0004C12N9/001C12N9/0069C12N9/1022C12N9/1029C12N9/1092C12N9/14C12N9/88C12N15/8261C12N15/8271C12N15/8275C12N15/8278C12N15/8289C12Y103/03004C12Y113/11027C12Y113/12019C12Y202/01006C12Y205/01019
Inventor 谢香庭丁德荣鲍晓明
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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