Four-primer molecular marker method for identifying ALS (acetolactate synthase) gene mutation generation of rice

Abstract

The invention relates to a four-primer molecular marker method for identifying ALS (acetolactate synthase) gene mutation generation of rice, and belongs to the field of biotechnology engineering. According to the variation of the rice ALS genes at 1642nd and 1643rd basic groups in a coding region from TG to AT, four specific molecular marker primers are designed and are added into a same PCR (polymerase chain reaction) system; different rice DNA (deoxyribonucleic acid) is subjected to amplification; electrophoresis detection is performed; DNA stripes with different sizes represent homozygotes and heterozygotes with different genotypes of the ALS. The method can be used for fast and accurately identifying the anti-ALS inhibitor type herbicide type gene variation occurring in the rice germplasm resources or breeding groups; in addition, the selection efficiency on the ALS-gene-mutation-containing homozygous genotypes can be improved in the breeding process; the breeding process of the anti-ALS inhibitor type herbicide rice varieties is accelerated.

Description

1. Technical field [0001] The invention relates to a four-primer molecular marker method for identifying ALS gene mutations in rice, which belongs to the field of biotechnology engineering and is specially used for the identification and variety breeding of rice variety resources resistant to ALS inhibitor herbicides. 2. Background technology [0002] With the continuous development of society and economy, a large number of rural labor force has been transferred. As an important way of light and simple rice cultivation, direct-seeding rice has been welcomed by the majority of farmers due to the obvious effect of saving labor and cost, and the planting area has expanded rapidly. Taking Jiangsu Province as an example, in 2008 the area of ​​direct-directed rice in the province reached 693,000 hm 2 , accounting for about 33% of the total rice area (Sun Tongqing et al., China Rice, 2014, 20(6): 5-9). In recent years, with the rapid promotion of machine transplanting technology a...

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Problems solved by technology

[0007] Technical problem: the present invention aims at the fact that traditional AS-PCR results have many false negatives, and heterozygous genotypes cannot be detected at one time, and KASP technology has high requirements for primers and detection equipment, and most domestic breeding units are currently difficult to apply, etc. Disadvantages: According to the mutations from TG to AT in the 1642nd and 1643rd bases of the ALS gene coding region between the rice Huanghuazhan mutant plant 1 and the wild type Huanghuazhan resistant to ALS inhibitor herbicides, four specific primers were designed and a one-step PCR method was used to quickly , Accurate identification of ALS gene mutations in rice, in order to achieve the purpose of resource identification and marker-assisted breeding of resistant herbicide-resistant varieties of ALS inhibitors

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