Miao Lixiang, Shou Senyan, Cai Jiayan, Jiang Fang, Zhu Zhujun, Li Hongbin
Department of Horticulture, College of Agriculture and Biotechnology, Zhejiang University, Hangzhou, China.
Mol Biol Rep. 2009 Mar;36(3):479-86. doi: 10.1007/s11033-007-9204-1. Epub 2007 Dec 23.
Tomato bacterial wilt (BW) incited by Ralstonia solanacearum is a constraint on tomato production in tropical, subtropical and humid regions of the world. In this paper, we present the results of a research aimed at the identification of PCR-based markers amplified fragment length polymorphism (AFLP) linked to the genes that confer resistance to tomato BW. To this purpose, bulked segregant analysis was applied to an F(2) population segregating for the BW resistant gene and derived from the pair-cross between a BW resistant cultivar T51A and the susceptible cultivar T9230. Genetic analysis indicated that tomato BW was conferred by two incomplete dominant genes. A CTAB method for total DNA extraction, developed by Murray and Thompson with some modifications was used to isolation the infected tomato leaves. Thirteen differential fragments were detected using 256 primer combinations, and two AFLP markers were linked to the BW resistance. Subsequently, the AFLP markers were converted to co-dominant SCAR markers, named TSCAR(AAT/CGA) and TSCAR(AAG/CAT). Linkage analysis showed that the two markers are on the contralateral side of TRSR-1. Genetic distance between TSCAR(AAT/CGA) and TRS-1 was estimated to 4.6 cM, while 8.4 cM between TSCAR(AAG/CAT) and TRS-1.
由青枯雷尔氏菌引起的番茄青枯病是世界热带、亚热带和湿润地区番茄生产的一大制约因素。在本文中,我们展示了一项研究的结果,该研究旨在鉴定与赋予番茄抗青枯病能力的基因连锁的基于聚合酶链式反应(PCR)的扩增片段长度多态性(AFLP)标记。为此,将混合分离群体分析法应用于一个因抗青枯病基因而分离的F₂群体,该群体来源于抗青枯病品种T51A与感病品种T9230的成对杂交。遗传分析表明,番茄青枯病由两个不完全显性基因控制。采用Murray和Thompson开发并经一些修改的十六烷基三甲基溴化铵(CTAB)法提取总DNA,用于分离感染青枯病的番茄叶片。使用256对引物组合检测到13个差异片段,其中两个AFLP标记与抗青枯病性状连锁。随后,将AFLP标记转化为共显性序列特征扩增区域(SCAR)标记,命名为TSCAR(AAT/CGA)和TSCAR(AAG/CAT)。连锁分析表明,这两个标记位于TRSR - 1的对侧。TSCAR(AAT/CGA)与TRS - 1之间的遗传距离估计为4.6厘摩,而TSCAR(AAG/CAT)与TRS - 1之间为8.4厘摩。