雄性和雌性中国甲鱼(Pelodiscussinensis)具有性别二态的生长模式,和男性有更高的商业价值,因为他们更大的尺寸和较厚的calipash。因此,开发性别特异性标记有利于研究中华黄曲霉的全雄性育种。这里,我们开发了一种准确有效的工作流程,用于使用ZW或XY性别确定系统筛选性别特异性序列。基于此工作流程,使用从头组装获得了2.23Gb和2.26Gb的雌性和雄性P.sinensis参考基因组。对齐和过滤后,最终鉴定了4.01Mb雌性特异性序列。随后,开发的七个性别特异性引物对初步准确率为100%,人口,和胚胎验证。P44,P45,P66,P67,P68和P69引物的条带的存在和不存在,以及PB1引物的两个和一个条带,表明胚胎在基因上是雌性和雄性,分别。NR和功能注释确定了几个性别决定的候选基因和相关途径,包括Ran,Eif4et,和Crkl基因,胰岛素信号通路和cAMP信号通路,分别。总的来说,我们的结果揭示了ZW型性别决定系统存在于中华黄曲霉中,并为筛选性别特异性标记提供了新的见解,性别控制育种,并对其性别决定机制进行了研究。
Male and female Chinese soft-shelled turtles (Pelodiscus sinensis) have sex-dimorphic growth patterns, and males have higher commercial value because of their larger size and thicker calipash. Thus, developing sex-specific markers is beneficial to studies on all-male breeding in P. sinensis. Here, we developed an accurate and efficient workflow for the screening of sex-specific sequences with ZW or XY sex determination systems. Based on this workflow, female and male P. sinensis reference genomes of 2.23 Gb and 2.26 Gb were obtained using de novo assembly. After aligning and filtering, 4.01 Mb female-specific sequences were finally identified. Subsequently, the seven developed sex-specific primer pairs were 100% accurate in preliminary, population, and embryonic validation. The presence and absence of bands for the primers of P44, P45, P66, P67, P68, and P69, as well as two and one bands for the PB1 primer, indicate that the embryos are genetically female and male, respectively. NR and functional annotations identified several sex-determining candidate genes and related pathways, including Ran, Eif4et, and Crkl genes, and the insulin signaling pathway and the cAMP signaling pathway, respectively. Collectively, our results reveal that a ZW-type sex-determination system is present in P. sinensis and provide novel insights for the screening of sex-specific markers, sex-control breeding, and the studies of the sex determination mechanism of P. sinensis.