Herbicide resistance protein and encoding genes thereof and application thereof

A resistance gene and herbicide technology, applied in genetic engineering, plant gene improvement, application, etc., can solve the problem of undiscovered 24DT02 herbicide resistance protein expression level, herbicide tolerance report, etc., to improve flexibility, The effect of high flexibility

Active Publication Date: 2013-04-24
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 24DT02 herbi...

Method used

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  • Herbicide resistance protein and encoding genes thereof and application thereof
  • Herbicide resistance protein and encoding genes thereof and application thereof
  • Herbicide resistance protein and encoding genes thereof and application thereof

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example 、24D

[0098] The first embodiment, the acquisition and synthesis of 24DT02 gene sequence

[0099] 1. Obtain the 24DT02 gene sequence

[0100] The amino acid sequence (298 amino acids) of the 24DT02 herbicide resistance protein is shown in SEQ ID NO: 2 in the sequence listing; the amino acid sequence (298 amino acids) encoding the 24DT02 herbicide resistance protein corresponding to said 24DT02 herbicide resistance protein is obtained according to the plant-preferred codon amino acids) nucleotide sequence (897 nucleotides), as shown in SEQ ID NO:1 in the sequence listing.

[0101] 2. Synthesize the above-mentioned 24DT02 nucleotide sequence

[0102] The 24DT02 nucleotide sequence (shown as SEQ ID NO: 1 in the sequence listing) was synthesized by Nanjing KingScript Biotechnology Co., Ltd.; the 5' of the synthesized 24DT02 nucleotide sequence (SEQ ID NO: 1) A SpeI restriction site is also connected to the end, and a KasI restriction site is also connected to the 3' end of the 24DT02 ...

no. 2 example

[0106] The second embodiment, construction of Arabidopsis recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0107] 1. Construction of Arabidopsis thaliana recombinant cloning vector DBN01-T containing 24DT02 nucleotide sequence

[0108] The synthetic 24DT02 nucleotide sequence was ligated into 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, which Build process such as 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; 24DT02 is 24DT02 Nucleotide sequence (SEQ ID NO: 1); MCS is multiple cloning site).

[0109] Then, the recombinant cloning vector DBN01-T was tra...

no. 3 example

[0125] The third embodiment, the acquisition of Arabidopsis plants transferred to 24DT02 nucleotide sequence

[0126] Wild-type Arabidopsis seeds were suspended in 0.1% agarose solution. Suspended seeds were stored at 4°C for 2 days to fulfill the need for dormancy to ensure simultaneous seed germination. Mix horse manure with vermiculite and subirrigate with water until moist, allowing the soil mixture to drain for 24 hours. The pretreated seeds were planted on the soil mixture and covered with a moisture hood for 7 days. Let the seeds germinate and keep the light intensity at 120-150μmol / m at constant temperature (22°C) and constant humidity (40-50%) 2 Plants were grown in the greenhouse under long-day conditions (16 h light / 8 h dark) of 2 sec. Start by watering the plants with Hoagland's nutrient solution, followed by deionized water, keeping the soil moist but not soggy.

[0127] Transform Arabidopsis using the flower soak method. Inoculate one or more 15-30ml precult...

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Abstract

The invention relates to herbicide resistance protein, encoding genes of the herbicide resistance protein and application of the herbicide resistance protein. The herbicide resistance protein comprises protein (a) which is composed of an amino acid sequence shown in a second sequence generator (SEQ) ID, or protein (b) which is derived by protein (a) and provided with herbicide resistance activity through replacement and /or deficiency and /or addition of one or more amino acids of the amino acid sequence in the protein (a), or protein (c) which is composed of an amino acid sequence which has at least 90 percent of sequence identity with the second SEQ ID. The herbicide resistance protein is particularly suitable for expression in plants, wide in herbicide resistance, and particularly used in phenoxy auxin herbicide.

Description

technical field [0001] The present invention relates to a herbicide resistance protein, its encoding gene and application, in particular to a protein with resistance to 2,4-D, its encoding gene and application. 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 resista...

Claims

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Application Information

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IPC IPC(8): C12N9/02C12N15/53C12N15/63C12N5/10C12N1/21C12N1/19C12N7/01A01K67/033C12N15/84A01H5/00A01G13/00
CPCA01N25/32C12N9/0069C12N15/8274C12Y113/11A01N37/46A01N39/04
Inventor 吴业春陶青丁德荣张成伟王登元黄金存王娜李彦勋刘晓艳鲍晓明
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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