Rape multiple ALS (acetolactate synthase) inhibitor type herbicide-resistant gene based on in-vitro site-specific mutagenesis and application

A technology of herbicide resistance and site-directed mutation, applied in application, genetic engineering, plant genetic improvement, etc., can solve problems such as heavy workload

Inactive Publication Date: 2017-08-18
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In view of the randomness of mutagenesis, a large number of seeds need to be screened, and the workload is relatively large

Method used

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  • Rape multiple ALS (acetolactate synthase) inhibitor type herbicide-resistant gene based on in-vitro site-specific mutagenesis and application
  • Rape multiple ALS (acetolactate synthase) inhibitor type herbicide-resistant gene based on in-vitro site-specific mutagenesis and application
  • Rape multiple ALS (acetolactate synthase) inhibitor type herbicide-resistant gene based on in-vitro site-specific mutagenesis and application

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Experimental program
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Effect test

Embodiment Construction

[0064] (1) Materials and methods:

[0065] 1 Material:

[0066]1.1 Herbicide-resistant Brassica napus strain M342 (see authorized patent ZL201310111739.5, a sulfonylurea herbicide-resistant gene and its application in Brassica napus), was induced by the Rapeseed Research Office of the Institute of Economic Crops, Jiangsu Academy of Agricultural Sciences through EMS. obtained, and studies have shown that the strain is resistant to sulfonylurea herbicides. Because of its ALS 3 A point mutation occurs at base 1667 of the gene, from the original G base to a T base, resulting in a mutation of the encoded protein sequence from tryptophan (W) to leucine (L) at position 556, Namely W556L. (see authorized patent ZL201310111739.5)

[0067] 1.2 Fast Pfu DNA polymerase, cloning vector Peasy-T1, Tag DNA polymerase, plasmid extraction, and gel recovery kits were purchased from Nanjing Biosky Biotechnology Co., Ltd. Recombinant cloning kit ClonExpressTM, PCR mix were purchased from Nanji...

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Abstract

The invention relates to a rape multiple ALS (acetolactate synthase) inhibitor type herbicide-resistant gene based on in-vitro site-specific mutagenesis and application, and belongs to the technical field of biology. The gene mALS3 is obtained by introducing a new mutant site into the ALS3 gene sequence of the herbicide-resistant rape M342, and the site is positioned at the 560th site of the ALS3 gene; the original gene is alanine with a C alkaline group at the 187th site, the C alkaline group is mutated into a T alkaline group, and valine is formed at the 187th site. The rape mALS3 gene with double mutation sites A187V and W556L is obtained through an in-vitro site-specific mutation technique. The double mutation sites are not reported in the literatures of the rape genome. After preliminary functional verification of transgenic arabidopsis, the rape mALS3 gene with double mutation sites has the advantages that the obvious resistance on multiple ALS inhibitor herbicides, such as imidazolamide, tribenuron-methyl, difluorosulfonamide and bispyribac-sodium, is realized; the resistance gene can be used for the culturing of herbicide-resistant crops.

Description

[0001] 1. Technical field [0002] The invention relates to a rape resistance multiple ALS inhibitor herbicide gene obtained based on in vitro site-directed mutation and its application, belonging to the field of biotechnology. [0003] 2. Background technology [0004] On the one hand, the growth of weeds in the field competes with crops for nutrients and growth space, which seriously affects the normal growth and development of crops; on the other hand, manual weeding is time-consuming and labor-intensive and increases labor costs. In modern agricultural production, more and more rely on chemical herbicides to control weeds. However, herbicides often cause varying degrees of damage to crops while killing weeds. Therefore, breeding new varieties of herbicide-resistant crops has important practical significance for the popularization and application of herbicides and the light and simplified cultivation of crops. [0005] Herbicides inhibit plant growth or kill plants primari...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N15/82A01H5/00
CPCC12N9/1022C12N15/8205C12N15/8274C12Y202/01006
Inventor 陈松浦惠明彭琦张洁夫胡茂龙付三雄陈锋周晓婴龙卫华高建芹张维王晓东
Owner JIANGSU ACAD OF AGRI SCI
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