安氏新银鱼(Neosalanx anderssoni)性别特异性位点筛选及性别鉴定方法开发  

SEX-SPECIFIC MOLECULAR MARKER SCREENING AND RAPID SEX IDENTIFICATION METHOD FOR NEOSALANX ANDERSSONI

作　　者：王心悦 邢腾飞 杨昊 李玉龙 刘进贤 WANG Xin-Yue;XING Teng-Fei;YANG Hao;LI Yu-Long;LIU Jin-Xian(Key Laboratory of Marine Ecology and Environmental Sciences,Institute of Oceanology,Chinese Academy of Sciences,Qingdao 266071,China;University of Chinese Academy of Sciences,Beijing 100049,China)

机构地区：[1]中国科学院海洋研究所海洋生态与环境科学重点实验室,山东青岛266071 [2]中国科学院大学,北京100049

出　　处：《海洋与湖沼》2025年第2期467-474,共8页Oceanologia Et Limnologia Sinica

基　　金：国家重点研发计划项目,2023YFC3108005号;国家自然科学基金项目,32370563号。

摘　　要：安氏新银鱼是我国重要的小型经济鱼类物种,具有较高的经济价值。但目前缺乏一种快速、经济有效的性别鉴定方法,极大地阻碍了其性别决定分子机制、性染色体演化历程的研究和水产育种与养殖技术方面的生产实践。本研究通过比较安氏新银鱼雌性与雄性的全基因组重测序数据,发现雌雄之间存在一个显著差异的基因组区域(chr02:2798574~5434239 bp)。基于测序深度差异分析筛选得到3个雌性特异性片段,长度分别为10 bp、167 bp和53 bp。在长为167 bp的雌性特异性位点附近设计引物,PCR扩增及凝胶电泳结果显示,雌性在该区域扩增产物具有2条带,雄性具有1条带,且雄性条带的亮度高于雌性两条带的亮度。该引物可以有效地区分安氏新银鱼的遗传性别,并分别在双洋港、秦皇岛已知性别的两个群体22个雌性和22个雄性个体中得到了验证。为了进一步验证研究结果的可靠性,分别从两个群体中随机选取雌雄个体样本的PCR产物进行测序分析,结果与预期相符,体现了该方法的有效性。研究结果也支持安氏新银鱼的性别决定系统为ZZ/ZW系统。本研究开发的方法可快速有效鉴定安氏新银鱼的遗传性别,并为该物种性别决定遗传机制及性染色体演化等后续研究提供理论支撑。The Neosalanx anderssoni is a small-sized economically valuable fish species in China.However,the lack of a rapid and cost-effective sex identification method has hindered research on the molecular mechanisms of sex determination,evolutionary history of sex chromosomes,and aquaculture breeding practices.Through comparison of whole-genome resequencing data between females and males,we identified a genome region with significant differences(chr02:2798574~5434239 bp).Three female-specific loci of lengths 10 bp,167 bp,and 53 bp were identified based on the analysis of sequencing depth differences.The female-specific 167 bp genomic region was chosen for design of sex specific primers.The results of traditional PCR amplification followed by gel electrophoresis revealed that females exhibited two distinct bands,whereas males displayed a single band with higher intensity than the two female bands.This method was further validated using 22 females and 22 males of known sex from Shuangyanggang and Qinhuangdao populations.To further validate the reliability of our research findings,we randomly selected PCR products of male and female individuals from two populations for sanger sequencing.The sequencing results were consistent with expectations,thereby strongly supporting our research conclusions.Our findings further demonstrated that the sex determination system of N.anderssoni was ZZ/ZW.These results not only enable accurate identification of the sex in N.anderssoni,but also provide valuable theoretical insights into the genetic mechanisms underlying sex determination and the evolutionary dynamics of sex chromosomes.

关 键 词：安氏新银鱼 雌性特异性位点 遗传性别鉴定 性别决定系统 引物开发 

分 类 号：Q319[生物学—遗传学]