Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China; STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou 515063, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266071, China.
Guangdong Provincial Key Laboratory of Marine Biotechnology, Shantou University, Shantou 515063, China; STU-UMT Joint Shellfish Research Laboratory, Shantou University, Shantou 515063, China.
Genomics. 2020 Jan;112(1):404-411. doi: 10.1016/j.ygeno.2019.03.003. Epub 2019 Mar 7.
In this study, we first identified male-specific SNP markers using restriction site-associated DNA sequencing, and further developed a PCR-based sex identification technique for Charybdis feriatus. A total of 296.96 million clean reads were obtained, with 114.95 and 182.01 million from females and males. After assembly and alignment, 10 SNP markers were identified being heterozygous in males but homozygous in females. Five markers were further confirmed to be male-specific in a large number of individuals. Moreover, two male-specific sense primers and a common antisense primer were designed, using which, a PCR-based genetic sex identification method was successfully developed and used to identify the sex of 103 individuals, with a result of 49 females and 54 males. The presence of male-specific SNP markers suggests an XX/XY sex determination system for C. feriatus. These findings should be helpful for better understanding sex determination mechanism, and drafting artificial breeding program in crustaceans.
在这项研究中，我们首先使用基于限制位点相关 DNA 测序的方法鉴定了雄性特异性 SNP 标记，然后进一步开发了一种基于 PCR 的 Charybdis feriatus 性别鉴定技术。共获得 2.9696 亿条清洁读取序列，其中雌性和雄性分别为 1.1495 亿条和 1.8201 亿条。组装和比对后，在雄性中鉴定出 10 个 SNP 标记为杂合子，而在雌性中则为纯合子。在大量个体中进一步证实了其中 5 个标记为雄性特异性。此外，设计了两个雄性特异性的有义引物和一个通用的反义引物，成功地开发了基于 PCR 的遗传性别鉴定方法，并用于鉴定 103 个个体的性别，其中 49 个为雌性，54 个为雄性。雄性特异性 SNP 标记的存在表明 Charybdis feriatus 存在 XX/XY 性别决定系统。这些发现有助于更好地理解甲壳动物的性别决定机制，并制定人工繁殖计划。
