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半滑舌鳎(Cynoglossus semilaevisGünther)为东北亚特有的名贵冷温性比目鱼类,为我国养殖业的新宠。半滑舌鳎雌鱼生长速度是雄性的2~3倍,若能实现单雌化养殖将大大提高养殖业的经济效益。本研究利用扩增片段长度多态性(AFLP)技术,应用64个引物组合,检测了半滑舌鳎(Cynoglossus semilaevisGünther)雌雄基因组DNA的多态性,筛选与半滑舌鳎性别相关的AFLP分子标记。实验经过3轮筛选和验证,4个引物组合扩增出7个雌性个体出现频率为100%的DNA片段,我们认为这7个标记是半滑舌鳎雌性特异的AFLP标记,分别命名为CseF382、CseF575、CseF783、CseF464、CseF136、CseF618和CseF305。同时,将标记CseF382成功转化为SCAR标记,测定了该标记的DNA序列,建立了半滑舌鳎遗传性别鉴定的PCR技术,为半滑舌鳎性别决定机制的研究和性别控制奠定了重要基础。
Cynoglossus semilaevis Günther is a rare cold-temperate flounder endemic to Northeast Asia, which is the new favorite of China’s aquaculture industry. Half-gliding tongue 鳎 female growth rate of male is 2 to 3 times, if the realization of single-female farming will greatly enhance the economic benefits of farming. In this study, polymorphism of male and female genomic DNA of Cynoglossus semilaevis Günther was detected by using the amplified fragment length polymorphism (AFLP) technique and 64 primer combinations. AFLP markers related to gender were also screened. After three rounds of screening and validation, the four primer combinations amplified 100 DNA fragments of seven females, and we think these seven markers are female-specific AFLP markers named CseF382, CseF575 and CseF783 , CseF464, CseF136, CseF618 and CseF305. At the same time, the CseF382 marker was successfully transformed into SCAR marker, and the DNA sequence of the marker was determined. The PCR technique was established for genetic determination of C. sinensis, which lays an important foundation for the study of sex determination mechanism and gender control of P. sinensis.