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当前位置: 首页 > 文章 > M342抗除草剂基因CAPS标记的开发与应用 江苏农业学报 2019,35 (2) 241-247
Position: Home > Articles > Development and application of the CAPS marker for herbicide-resistant gene in M342 Jiangsu Journal of Agricultural Sciences 2019,35 (2) 241-247

M342抗除草剂基因CAPS标记的开发与应用

作  者：
程丽;胡茂龙;浦惠明;龙卫华;高建芹;陈锋;周晓婴;张维;陈松;张洁夫
单  位：
江苏省农业科学院经济作物研究所/农业部长江下游棉花与油菜重点实验室/国家油料作物改良中心南京分中心/江苏省现代作物生产协同创新中心
关键词：
油菜;磺酰脲类除草剂;乙酰乳酸合酶;酶切扩增多态性序列(CAPS)标记
摘  要：
M342是利用EMS诱变定向选育获得的甘蓝型油菜抗磺酰脲类除草剂新种质.通过M342抗性基因与敏感型油菜的ALS基因序列分析结果表明,M342的BnALS3基因存在1处SNP突变,导致编码的蛋白质第556位色氨酸突变为亮氨酸,该突变导致基因序列对BsrD I内切酶消化的差异.为此设计了8条引物,从中筛选到引物对SU54F/SU58R在不同油菜品种中可以特异性扩增出目的片段,该片段经BsrD I酶切分型正确,从而开发了检测M342中抗性基因BnALS3R的CAPS标记.运用该标记对除草剂敏感型油菜、纯合抗性油菜及杂合抗性油菜BnALS3的基因型进行验证,并应用该标记对F2、BC1分离群体进行检测.结果表明,该标记验证的基因型结果与测序结果一致.F2分离群体有Als3Als3、Als3als3、als3als33种基因型,BC1分离群体有Als3als3、als3als32种基因型,与表型鉴定结果一致,遵循单基因遗传分离规律,表明该标记可以应用于抗性基因的检测.CAPS标记的获得为抗除草剂油菜的分子标记辅助选择育种奠定了基础.
译  名：
Development and application of the CAPS marker for herbicide-resistant gene in M342
作  者：
CHENG Li;HU Mao-long;PU Hui-ming;LONG Wei-hua;GAO Jian-qin;CHEN Feng;ZHOU Xiao-ying;ZHANG Wei;CHEN Song;ZHANG Jie-fu;Institute of Industrial Crops, Jiangsu Academy of Agricultural Sciences/Key Laboratory of Cotton and Rapeseed (Nanjing), Ministry of Agriculture/Nanjing Sub-center, National Center of Oil Crops Improvement/Jiangsu Collaborative Innovation Centre for Modern Crop Production;
关键词：
rapeseed(Brassica napus L.);;sulfonylurea herbicides;;acetolactate synthase;;cleaved amplified polymorphic sequence(CAPS) marker
摘  要：
M342, a sulfonylurea-resistant germplasm in Brassica napus L., was developed and obtained by ethylmethane sulfonate seed treatment. According to the analysis of the cloned ALS gene sequences between M342 and herbicide-sensitive rapeseed, one single nucleic acid polymorphism found in BnALS3 of M342 led to substitution of tryptophan to leucine at site 556 of ALS, giving rise to the difference of digestion with BsrD I between ALS genes. Thus, eight primers were designed, from which a pair of primers SU54 F/SU58 R amplifying the targeted fragments specifically in different rapeseed varieties was selected. The genotyping results of the PCR fragments digested with BsrD I were accurately got, and the CAPS marker detecting the resistant gene of BnALS3 R in M342 was developed. Then the genotypes of BnALS3 were validated in herbicide-sensitive, homozygous and heterozygous resistant rapeseed, and the F_2 and BC_1 populations were detected by CAPS marker. A perfect correlation between the observed CAPS genotyping results, and the sequencing alignment results for the different lines was found. There were three genotypes of Als3 Als3, Als3 als3 and als3 als3 in F_2 population, two genotypes of Als3 als3 and als3 als3 in BC_1 population, which was in accordance with the results of phenotype identification. The segregation followed the genetic law for a single gene model. The results confirmed that the CAPS marker could be applied to the detection of the resistant gene. The acquisition of CAPS marker lays a foundation for the marker-assisted selection breeding of herbicide-resistant rapeseed.

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