AFLP and SCAR markers linked to the suppressor gene (Rf) of a dominant genetic male sterility in rapeseed (Brassica napus L.)
AFLP and SCAR markers linked to the suppressor gene (Rf) of a dominant genetic male sterility in rapeseed (Brassica napus L.)
复制标题
DOI：
10.1007/s10681-006-9162-z
复制
发表时间：
2006-10-01
期刊：
影响因子：
1.9
通讯作者：
He, Qingbiao
中科院分区：
文献类型：
--
作者：
Hong, Dengfeng;Wan, Lili;He, Qingbiao
Rs 1046 AB是一个显性遗传雄性不育基因（Ms）的真正育种系，但它是雄性可育和不育个体的混合物（两型系），因为它分离显性抑制基因（Rf）。该系统为甘蓝型油菜杂交育种提供了一个很有前途的CMS系统的替代方案。为了确定与rf基因连锁的分子标记，从Rs 1046 AB的不育个体（MsMsMsRfrf）和可育个体（MsMsRfrf）之间的杂交的近等基因系（NIL）群体进行扩增片段长度多态性（AFLP）分析，与近等基因系（NIL）比较和混合分离分析（BSA）相结合。从2,816对AFLP引物中筛选出6个在可育群体和不育群体间以及群体个体间表现多态性的片段。连锁分析表明，这6个AFLP标记与Rf基因紧密连锁，且均分布在同一侧。Rf基因与标记之间的最小遗传距离为0.7cM。由于AFLP标记不适合大规模应用于标记辅助选择（MAS），我们的目标是开发一种快速，廉价和可靠的PCR检测。因此，四个最接近的AFLP标记中的三个被直接转换为序列特征扩增区（SCAR）标记。对于另一个标记，通过PCR Walking分离相邻序列后，成功地获得了相应的SCAR标记。Rf基因的SCAR标记将极大地促进未来利用显性核不育产生杂交品种的育种计划。
Rs1046AB is a line which is true breeding for a dominant genetic male sterility gene (Ms) but which is a mixture of male fertile and sterile individuals (a two-type line) because it is segregating for a dominant suppressor gene (Rf). This system provides a promising alternative to the CMS system for hybrid breeding in Brassica napus. In order to identify molecular markers linked to the rf gene, a near-isogenic line (NIL) population from the cross between a sterile individual (MsMsrfrf) and a fertile individual (MsMsRfrf) in Rs1046AB was subjected to amplified fragment length polymorphism (AFLP) analysis, with a combination of comparing near isogenic lines (NILs) and bulked segregant analysis (BSA). From 2,816 pairs of AFLP primers, six fragments showing polymorphism between the fertile and sterile bulks as well as the individuals of the bulks were identified. Linkage analysis indicated that the six AFLP markers are tightly linked to the Rf gene and all are distributed on the same side. The minimum genetic distance between the Rf gene and a marker was 0.7 cM. Since the AFLP markers are not suitable for large-scale application in MAS (marker-assisted selection), our objective was to develop a fast, cheap and reliable PCR-based assay. Consequently, three of the four closest AFLP markers were converted directly to sequence characterized amplified region (SCAR) markers. For the other marker a corresponding SCAR marker was successfully obtained after isolating the adjacent sequences by PCR Walking. The available SCAR markers of the Rf gene will greatly facilitate future breeding programs using dominant GMS to produce hybrid varieties.