A functional genomics resource for Brassica napus: development of an EMS mutagenized population and discovery of FAE1 point mutations by TILLING
A functional genomics resource for Brassica napus: development of an EMS mutagenized population and discovery of FAE1 point mutations by TILLING
复制标题
甘蓝型油菜的功能基因组学资源：EMS 诱变群体的开发以及 TILLING 发现的 FAE1 点突变。
DOI：
10.1111/j.1469-8137.2008.02619.x
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发表时间：
2008-01-01
期刊：
影响因子：
9.4
通讯作者：
Meng, Jinling
中科院分区：
文献类型：
--
作者：
Wang, Nian;Wang, Yajie;Meng, Jinling
两个半冬性油菜品种的甲基磺酸乙酯(EMS)突变体群体。构建了高突变载量的宁油7号，为甘蓝型油菜功能基因组学研究和油菜新等位变异育种提供了新的平台。从M2群体中对突变体进行正向遗传筛选，发现了大量新的表型。反向遗传筛选的重点是控制油菜种子芥酸合成的潜在多基因FAE1(脂肪酸拉长1)。对甘蓝型油菜BAC文库进行了筛选，并对参考作图群体(TNDH)中的基因座进行了定位，结果表明在甘蓝型油菜A和C基因组上各有1个FAE1基因。在基因组中存在两个或更多非常相似的平行基因拷贝的情况下，一种新的程序被证明可以识别新的突变。该程序涉及到单株植物的耕作，使用现有的SNPs作为阳性对照，并能够根据旨在同时扩增两个FAE1类似物的引物对区分新的突变。该程序应用于1344株M2植株，鉴定出19个突变，其中3个突变功能受损，种子芥酸含量降低。
Two ethylmethanesulfonate (EMS) mutant populations of the semi-winter rapeseed cv. Ningyou7 were constructed with high mutant load, to provide a TILLING platform for functional genomics in Brassica napus, and for introduction of novel allelic variation in rapeseed breeding. Forward genetic screening of mutants from the M2 populations resulted in identification of a large number of novel phenotypes. Reverse genetic screening focused on the potentially multi-paralogous gene FAE1 (fatty acid elongase1), which controls seed erucic acid synthesis in rapeseed. A B. napus BAC library was screened, and loci in a reference mapping population (TNDH) were mapped to conclude that there are two paralogous copies of FAE1, one on each of the B. napus A and C genomes. A new procedure is demonstrated to identify novel mutations in situations where two or more very similar paralogous gene copies exist in a genome. The procedure involves TILLING of single plants, using existing SNPs as a positive control, and is able to distinguish novel mutations based on primer pairs designed to amplify both FAE1 paralogues simultaneously. The procedure was applied to 1344 M2 plants, with 19 mutations identified, of which three were functionally compromised with reduced seed erucic acid content.