DataSheet_1_Application of SNP in Genetic Sex Identification and Effect of Estradiol on Gene Expression of Sex-Related Genes in Strongylocentrotus intermedius.zip

Sea urchin (Strongylocentrotus intermedius) is an economically important mariculture species in Asia, and its gonads are the only edible part. The efficiency of genetic breeding in sea urchins is hampered due to the inability to distinguish gender by appearance. In this study, we first identified a sex-associated single nucleotide polymorphism (SNP) by combining type IIB endonuclease restriction site-associated DNA sequencing (2b-RAD-seq) and genome survey. Importantly, this SNP is located within spata4, a gene specifically expressed in male. Knocking down of spata4 by RNA interference (RNAi) in male individuals led to the downregulation of other conserved testis differentiation-related genes and germ cell marker genes. We also revealed that sex ratio in this validated culture population of S. intermedius is not 1:1. Moreover, after a 58-day feeding experiment with estradiol, the expression levels of several conserved genes that are related to testis differentiation, ovary differentiation, and estrogen metabolism were dynamically changed. Taken together, our results will contribute toward improving breeding efficiency, developing sex-controlled breeding, and providing a solid base for understanding sex determination mechanisms in sea urchins.

海胆（强脊海胆）作为亚洲重要的经济养殖物种，其生殖腺是其唯一可食用部分。由于无法通过外观区分性别，海胆的遗传育种效率受到限制。在本研究中，我们首先通过结合II型B内切酶限制性位点关联DNA测序（2b-RAD-seq）和基因组调查，鉴定了一种与性别相关的单核苷酸多态性（SNP）。值得注意的是，该SNP位于spata4基因内部，该基因仅在雄性中特异性表达。通过RNA干扰（RNAi）在雄性个体中敲低spata4，导致其他保守的睾丸分化相关基因和生殖细胞标记基因的下调。此外，我们还揭示了在验证的文化种群中，S. intermedius的性别比例并非1:1。在经过58天的雌二醇喂养实验后，与睾丸分化、卵巢分化和雌激素代谢相关的多个保守基因的表达水平发生了动态变化。综上所述，我们的研究结果将有助于提高育种效率，开发性别控制育种，并为理解海胆性别决定机制提供坚实的基础。