Brassica napus mutator gene for resisting imidazolone weedicide and application thereof

A technology of imidazolinone-resistant and brassica napus, applied in the fields of application, plant genetic improvement, enzymes, etc., can solve the problems that brassica napus is resistant to imidazolinone herbicides that have not been disclosed or published, and achieve improved tolerance Effect

Active Publication Date: 2011-01-05
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

But so far, there has not been published or published the imidazolinone herbicide-resistant gene and its application in Brassica napus in China.

Method used

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  • Brassica napus mutator gene for resisting imidazolone weedicide and application thereof
  • Brassica napus mutator gene for resisting imidazolone weedicide and application thereof
  • Brassica napus mutator gene for resisting imidazolone weedicide and application thereof

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Effect test

Embodiment Construction

[0041] All methods in the following implementations are conventional methods unless otherwise specified.

[0042] (1) PCR Cloning and Sequencing of the Mutant Nucleic Acid Sequence of Brassica napus Acetolactate Synthase Gene

[0043] At the seedling stage, fresh and tender leaves of mutant M9, a spontaneous rapeseed mutant strain resistant to imidazolinone herbicides (publicly known and public, see Gao Jianqin et al., Journal of Plant Genetic Resources, 2010, 11(3): 369-373), were taken at -70°C Save for later. The total DNA extraction method of rapeseed genome was improved according to the CTAB method proposed by Murray et al. (1980). Proceed as follows:

[0044] (1) Put about 0.2g leaves in a mortar, add liquid nitrogen to grind, transfer to a 1.5ml centrifuge tube; (2) add 700μl extraction buffer (2% CTAB, 100mM Tris pH8.0, 20mM EDTA pH8 .0, 1.4M NaCl, add 1% β-Me before use), mix well, 65 ℃ water bath for 1.5h; (3) cool on ice, centrifuge at 12000rpm, 5min; (4) absorb...

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Abstract

The invention discloses a brassica napus mutator gene for resisting imidazolone weedicide and application thereof, relating to the field of plant molecule breeding. Primer pairs are utilized to carry out PCR (Polymerase Chain Reaction) proliferation by using a mutant M9 of brassica napus for resisting imidazolone weedicide as a material to obtain a mutator gene of the mutant with the sequence number of SEQ ID NO.1; and proteins are acquired by translation of the gene with an amino acid sequence of SEQ ID No.2. By applying the conventional breeding method of crossbreeding plants, backcrossing plants and the like, the gene is introduced into other napus varieties or variety series having no resistance to the imidazolone weedicide, so as to improve the durability on the imidazolone weedicide by introducing ALS gene mutation nucleotide sequence target varieties or variety series.

Description

technical field [0001] The invention relates to the field of plant molecular breeding, in particular to a mutant gene nucleic acid sequence of brassica napus encoding acetolactate synthase and capable of establishing imidazolinone herbicide resistance and its application. Background technique [0002] Imidazolinone herbicides are a class of broad-spectrum, low-consumption, low-toxicity to mammals and environmentally friendly herbicides successfully developed by American Cyamide Company in the early 1980s. The key enzyme of synthesis, acetolactate synthase (ALS), also known as acetohydroxyacid synthase (AHAS), causes the biosynthesis of isoleucine, valine and leucine to be blocked, and destroys protein synthesis in plants , causing the plants to yellow, inhibit growth, and finally die gradually. ALS is responsible for catalyzing two parallel reactions, catalyzing the condensation of two molecules of pyruvate to form 2-acetolactate to release CO 2 , finally generate valine a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/52C12N9/00A01H1/02A01H4/00A01G7/06
Inventor 胡茂龙浦惠明戚存扣张洁夫高建芹龙卫华陈松陈新军陈峰顾慧
Owner JIANGSU ACAD OF AGRI SCI
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