dCAPS (derived cleaved amplified polymorphic sequences) molecular marking method for rapidly screening imidazolinone herbicide resistant rice

Abstract

The invention discloses a dCAPS (derived cleaved amplified polymorphic sequences) molecular marking method for rapidly screening imidazolinone herbicide resistant rice. Sequencing comparison proves that mutation of amino acid at a 627 locus of each ALS (Acetolactate synthase) gene into N (Asparagine) is a primary cause of resistance of 'Jinjing 818' to imidazolinone herbicides, the ALS gene carrying a resistance locus is named as ALS627N, and the ALS gene without carrying the locus is named as ALS627S. According to DNA (deoxyribonucleic acid) sequence differences, a novel functional markerdC-ALS-627 is developed. The marker can effectively distinguish ALS627N homozygotic type, ALS627S homozygotic type and hemizygous lines. In addition, herbicide spraying tests prove that the ALS627N is a dominant herbicide resistant gene. Rice varieties in different regions are detected by the marker to prove that the marker can be widely used for marker-assisted breeding of the ALS627N gene, canaccelerate culture of new imidazolinone herbicide resistant varieties and has important practical application values.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for breeding new rice varieties resistant to imidazolinone herbicides by using functional molecular markers dCAPS markers. Background technique [0002] Acetolactate synthase (ALS) inhibitors are a class of herbicides that are widely used now. They have a wide spectrum of herbicides, low toxicity and high efficiency, strong selectivity, and are very safe to humans and animals (Zhang Yunyue et al., Weed Science, 2013, 31(2):1-5). Through large-scale mutagenesis screening, foreign scholars took the lead in obtaining innovative rice germplasm resistant to ALS inhibitors, and bred and promoted corresponding herbicide-resistant rice varieties (Sudianto E, et al., Crop Prot, 2013, 49:40-51 ). However, with the single application of a large number of herbicides of the same type, resistant weeds have been found in many planting fields (BusconiM, et al., Plant Biol, 201...

AI Technical Summary

Problems solved by technology

In order to accelerate the application of these germplasms in the breeding of new rice varieties resistant to ALS inhibitor herbicides, some scholars have developed functional markers for ALS mutants in these varieties, such as: Chen Tao et al. based on the four-primer amplification hindered mutation system The functional marker of the ALS mutant gene in Huanghuazhan M-1 was developed by PCR principle (Chen Tao et al., China Rice Science, 2018, 1(1):137-145). This marker was developed for the 548th amino acid of the ALS gene. It is suitable for the detection of mutation sites in Jinjing 818 and related derivative materials; Wang Fangquan et al. developed the ALS mutant gene functional marker AS-ALS in Jinjing 818 by using the principle of allele-specific PCR (Wang Fangquan et al., Acta Crops, 2018, 44 (3):324-331), however, in practice, the detection results of such markers are often false negative, and the one-time detection of heterozygous genotypes is not possible

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