对辣椒抗性基因Me3表现毒性的南方根结线虫 群体的SCAR分子标记.pdfVIP

中国农业科学 2013,46(5):943-949 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2013.05.009 对辣椒抗性基因Me3 表现毒性的南方根结线虫 群体的SCAR 分子标记 王 刚，李二峰，茆振川，谢丙炎，冯东昕 （中国农业科学院蔬菜花卉研究所农业部园艺作物生物学与种质创制重点实验室，北京100081） 摘要：【目的】研究南方根结线虫（Meloidogyne incognita）Me3 毒性变异群体的分子标记，用快速有效的 分子方法来鉴定毒性变异的发生。【方法】以南方根结线虫的无毒群体以及抗性基因Me3 毒性群体为材料，用根据 南方根结线虫基因组设计的100 对引物和19 对来自文献的引物进行扩增，筛选出在3 个种群中扩增的差异条带， 设计SCAR 引物，并建立多重PCR 反应体系。【结果】得到了7 对能在3 个群体稳定扩增出差异条带的多态性引物， 把其中的2 个位点开发为SCAR 标记，能够区分开3 个线虫群体。多重PCR 反应体系在无毒群体和Me3 毒性群体中 分别扩增出1 条999 bp 和1 条629 bp 的条带，在混合群体扩增出999 和629 bp 的2 条带。【结论】成功开发了 对Me3 表现毒性的南方根结线虫变异群体的分子标记，可用多重PCR 体系一次性鉴定Me3 毒性变异的发生。 关键词：南方根结线虫；分子标记；SCAR；毒性变异 SCAR Molecular Markers Correlated with Populations of Meloidogyne incognita Virulent to Resistance Gene Me3 WANG Gang, LI Er-feng, MAO Zhen-chuan, XIE Bing-yan, FENG Dong-xin (Key Laboratory of Horticultural Crops Biology and Genetic Improvement, Ministry of Agriculture, Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081) Abstract: 【Objective 】Molecular markers of virulent populations against Me3 in Meloidogyne incognita was studied in order to detect the virulence mutation rapidly and effectively. 【Method 】Root-knot nematode populations including avirulent population, populations overcoming resistant gene Me3 and the mixed group of the two population were used as experiment materials, polymerase chain reaction was done with 100 primer pairs designed according to M. incognita genome and 19 pairs reported in literature to screen specific band among three populations. And subsequently SCAR primers were designed and a multiplex PCR reaction system was built. 【Result 】Seven primer p