基于遗传连锁图谱筛选虾夷扇贝性别相关AFLP分子标记的方法

摘要

Patinopecten yessoensis is one of the most important germplasm resources in scallop breeding in north coast of China. It has male, female and hermaphrodite phenotypes. A rising hermaphroditism percentage in cultured populations was observed in gonochorism P. yessoensis. The previous studies indicated that hermaphroditisms were intermediate states of sex-reversal, and that the development of gonad was simultaneous in some hermaphroditic scallops. The hemaphroditisms could provide useful materials for inbred line construction and genetic improvement of scallop. AFLP (amplified fragment length polymorphism) markers have been used to construct the genetic linkage maps in the scallop P. yessoensis by several research groups since 2009. The sex-related AFLP molecular markers were utilized to screen female, male and hermaphrodite P. yessoensis from samples that were collected from March to May, 2011–2013. Eighteen AFLP primer-combinations were chosen to screen sex-related bands and 8 primer-combinations were sex-specific markers. Three of eight markers (EbMc, EjMf and EiMk) had a good repeatability. Hermaphrodite-specific bands were found in four primer-combinations and female-specific bands were detected in five primer-combinations. The present results indicate that the method of sex-related molecular markers screening based on the highly dense genetic linkage map was practical and feasible, and that the hermaphrodism is probably an independent group since the sex-related molecular markers in hermaphrodite genomic DNA were apparently different from those of the female and male. The method of sex-related markers screening and the sex-related bands screened may contribute to the efficiency improvement of sex control and breeding, and help to identify the hermaphrodite and then induce the female-and male-gametes to mature synchronously to construct self-bred-lines.%基于近年来他人构建的遗传连锁图谱，对2011–2013年2–5月期间采集的雌性、雄性及雌雄同体虾夷扇贝进行性别相关AFLP分子标记筛选。从雄性和雌性遗传连锁图谱上选取合计18对引物组合，经3批次试验，有8对引物扩增出性别相关的条带，其中EbMc、EjMf和EiMk重复性好，有4对引物扩增出雌雄同体特异性条带，5对引物扩增出雌性特异性条带。结果表明，基于高密度遗传连锁图谱筛选性别相关分子标记的方法简单可行，雌雄同体的基因组DNA与雌性和雄性存在显著差异，可能是一个单独的群体。