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Primers for detecting anti-sulfonylurea herbicide gene BnALS3R of cabbage type rape and application of primer

A Brassica napus, sulfonylurea-resistant technology, applied in application, plant genetic improvement, DNA/RNA fragment, etc., can solve the problems of improper use of herbicides, spraying, unevenness, etc., and achieves easy operation, accurate and reliable results. , the effect that is easy to judge

Active Publication Date: 2015-07-22
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

During the herbicide identification process of each segregated generation, if the herbicid

Method used

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  • Primers for detecting anti-sulfonylurea herbicide gene BnALS3R of cabbage type rape and application of primer
  • Primers for detecting anti-sulfonylurea herbicide gene BnALS3R of cabbage type rape and application of primer
  • Primers for detecting anti-sulfonylurea herbicide gene BnALS3R of cabbage type rape and application of primer

Examples

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Effect test

Embodiment 1

[0044] Example 1 The development process of the BnALS3R molecular marker for the detection of sulfonylurea herbicide resistance gene in Brassica napus

[0045] (1) Development of AS-PCR molecular markers for detection of rapeseed sulfonylurea herbicide resistance gene BnALS3R

[0046] The sulfonylurea herbicide resistance gene BnALS3R of Brassica napus was cloned from the mutant M342 induced by EMS (see Pu Huiming et al., patent application number 201310054645.9). Compared with the acetolactate synthase III gene of wild-type N131, the mutant BnALS3 gene had a single-base mutation, that is, +1667 guanine nucleotides (G) downstream of the first nucleotide of the start codon Mutated into a thymine nucleotide (T) (Hu Maolong et al., ZL201310111739.5, a sulfonylurea herbicide resistance gene and its application in Brassica napus), and related genes can be developed based on the single base variation of this site Molecular marker detection resistance gene BnALS3R.

[0047] First, ...

Embodiment 2

[0073] Example 2 Detection and Identification of Molecular Marker BsrDI-ALS3R in Segregation Populations Against Resistance Gene BnALS3R Genotype

[0074] In order to verify the detection effect of molecular marker BsrDI-ALS3R on BnALS3R genotypes resistant to sulfonylurea herbicides in Brassica napus, we tested it in three types of BnALS3R genotype isolates. The F2 population was obtained by selfing the F1 of the MICMS double-low restorer line N221 and M342; the BC1 population was obtained by backcrossing the F1 of the N221 and M342 cross with N221; The herbicide Rapeseed M342 was obtained by backcrossing. The leaves of the rapeseed plants were taken at the seedling stage, and the genome DNA was extracted for PCR amplification. DNA extraction, PCR reaction, enzyme digestion reaction, and electrophoresis detection methods are the same as above. The test results showed that there were three genotype plants in the F2 population ( image 3 ), that is, a homozygous resistant si...

Embodiment 3

[0075] Example 3 Breeding herbicide-resistant MICMS restorer lines using molecular marker-assisted selection

[0076] Breeding herbicide-resistant hybrid rapeseed can solve the current problem of chemical weed control of dicotyledonous weeds in rapeseed fields, saving labor and cost, and the herbicide-resistant trait can also be used as a marker trait in hybrid rapeseed production, provided that the resistant The herbicide traits were imported into the MICMS restorer lines. The MICMS restorer line resistant to sulfonylurea herbicides can be rapidly bred through the marker-assisted selection technique using the molecular marker BsrDI-ALS3R of the present invention. We tested the resistance genes of the plants in the F2 population obtained by selfing of three kinds of F1 (N221×M342, N340×M342 and N341×M342), and bagged the fertile individual plants with a molecular weight band of 766bp. F3 seeds containing homozygous sulfonylurea herbicide resistance gene BnALS3R were obtained....

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Abstract

The invention discloses primers for detecting an anti-sulfonylurea herbicide gene BnALS3R of cabbage type rape and the application of the primers, and belongs to the field of molecular breeding. A method for detecting the anti-sulfonylurea herbicide gene BnALS3R of the cabbage type rape by use of the primers SEQ ID NO.1 and SEQ ID NO.2 comprises the following steps: taking the total DNA of the cabbage type rape as a template and adding a primer for PCR amplification; performing restriction enzyme BsrDI digestion on the PCR product; differentiating homozygous rape containing the resistance gene BnALS3R, hybrid rape containing the resistance gene BnALS3R, and homozygous rape not containing the resistance gene BnALS3R by use of the enzyme digestion product. According to the primers, whether rape planets contain the resistance gene BnALS3R can be detected quickly and accurately, and whether the contained anti-herbicide gene is a homozygous type or a hybrid type also can be detected accurately, and therefore, the breeding objective of breeding a new anti-herbicide rape variety can be achieved quickly and effectively.

Description

technical field [0001] The invention relates to the field of molecular breeding, in particular to a primer and application for detecting the sulfonylurea herbicide resistance gene BnALS3R of Brassica napus. Background technique [0002] In recent years, with the acceleration of industrialization and urbanization in our country, a large number of rural labor force has been transferred to cities and towns. Simplification and mechanization have become the main direction of the development of rapeseed industry. Effective control of field weeds is an important technical link to realize the lightness, simplification and mechanization of rapeseed. Chemical weeding is currently the main means of controlling weeds in farmland. However, the herbicides currently used in rapeseed production can only effectively control monocotyledonous weeds, and there is a lack of safe and efficient herbicides for controlling most dicotyledonous broad-leaved weeds. The area of ​​chemical weeding in ra...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCA01H1/04C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 胡茂龙浦惠明龙卫华高建芹张洁夫陈松陈锋付三雄周晓婴张维彭琦王晓东戚存扣
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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