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Herbicide resistance protein, its encoding gene and use

A resistance gene and herbicide technology, applied in genetic engineering, plant gene improvement, application, etc., can solve the problems of reporting herbicide tolerance without finding the expression level of AADM3 herbicide resistance protein, to achieve increased flexibility, The effect of high flexibility

Active Publication Date: 2016-05-18
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the expression level of AADM3 herbicide resistance protein in plants and its tolerance to herbicides

Method used

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  • Herbicide resistance protein, its encoding gene and use
  • Herbicide resistance protein, its encoding gene and use
  • Herbicide resistance protein, its encoding gene and use

Examples

Experimental program
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Effect test

no. 1 example

[0093] The first embodiment, the acquisition and synthesis of AADM3 gene sequence

[0094] 1. Obtain the AADM3 gene sequence

[0095] The amino acid sequence (292 amino acids) of the AADM3 herbicide resistance protein is shown in SEQ ID NO: 2 in the sequence table; the amino acid sequence (292 amino acids) encoding the AADM3 herbicide resistance protein was obtained according to the plant-preferred codons ) Nucleotide sequence (879 nucleotides), as shown in SEQ ID NO: 1 in the sequence listing.

[0096] 2. Synthesize the above AADM3 nucleotide sequence

[0097] The AADM3 nucleotide sequence (shown in SEQ ID NO: 1 in the sequence list) was synthesized by Nanjing GenScript Biotechnology Co., Ltd.; the 5'end of the synthesized AADM3 nucleotide sequence (SEQ ID NO: 1) is also connected SpeI restriction site, the 3'end of the AADM3 nucleotide sequence (SEQ ID NO:1) is also connected with a KasI restriction site.

no. 2 example

[0098] The second embodiment, the construction of Arabidopsis recombinant expression vector and the transformation of recombinant expression vector into Agrobacterium

[0099] 1. Construct the Arabidopsis thaliana recombinant cloning vector DBN01-T containing the AADM3 nucleotide sequence

[0100] Connect the synthesized AADM3 nucleotide sequence to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and proceed according to the instructions of Promega product pGEM-T vector to obtain the recombinant cloning vector DBN01-T. The build process is like figure 1 As shown (where Amp represents the ampicillin resistance gene; f1 represents the origin of replication of phage f1; LacZ is the LacZ start codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; AADM3 is the AADM3 nucleoside Acid sequence (SEQIDNO:1); MCS is the multiple cloning site).

[0101] Then the recombinant cloning vector DBN01-T was transformed into E. coli T1 competent cells (Tran...

no. 3 example

[0111] The third embodiment, the acquisition of Arabidopsis plants transformed with AADM3 nucleotide sequence

[0112] The wild-type Arabidopsis seeds were suspended in a 0.1% agarose solution. The suspended seeds were stored at 4°C for 2 days to complete the need for dormancy to ensure synchronous germination of the seeds. Mix horse manure soil with vermiculite and irrigate it with water until it is wet, and drain the soil mixture for 24 hours. The pretreated seeds were planted on the soil mixture and covered with a moisturizing cover for 7 days. Let the seeds germinate and the light intensity is 120-150μmol / m at constant temperature (22℃) and humidity (40-50%) 2 Plants are cultivated in a greenhouse under the conditions of long daylight (16 hours of light / 8 hours of darkness) in seconds. Begin to irrigate the plants with Hoagland nutrient solution and then irrigate with deionized water to keep the soil moist but not wet.

[0113] Use flower soaking method to transform Arabidop...

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Abstract

The invention relates to a herbicide-resistant protein as well as an encoding gene and application thereof. The herbicide-resistant protein comprises: (a) a protein constituted by an amino acid sequence shown as SEQ ID NO: 2, or (b) a protein which is obtained by performing substitution and / or deletion and / or addition of one or more amino acid residues on the amino acid sequence in (a), has herbicide resisting activity and is derived from (a). The herbicide-resistant protein is particularly suitable for expression in plants, and is highly resistant to herbicides, especially a phenoxyl auxin herbicide.

Description

Technical field [0001] The present invention relates to a herbicide resistance protein, its coding gene and its use, in particular to a protein with 2,4-D resistance, its coding gene and its use. Background technique [0002] Weeds can quickly deplete valuable nutrients needed by crops and other purpose plants in the soil. There are currently many types of herbicides used to control weeds. One particularly popular herbicide is glyphosate. Crops that are resistant to glyphosate have been developed, such as corn, soybeans, cotton, sugar beets, wheat, and rice. Therefore, glyphosate can be sprayed on fields where glyphosate resistant crops are grown to control weeds without significantly damaging the crops. [0003] Glyphosate has been widely used globally for more than 20 years, which has led to excessive dependence on glyphosate and glyphosate-tolerant crop technologies, and is naturally more tolerant to glyphosate in wild weed species or has developed Plants that exhibit resista...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/63C12N15/84C12N5/10C12N1/13C12N1/19C12N1/21C12N7/01A01K67/033A01H5/00A01G13/00
CPCC12N9/0071C12N15/8209C12N15/8274
Inventor 陶青吴业春谢香庭丁德荣
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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