中华鲟性别相关的扩增片段长度多态性分子标记筛选
Screening of Sex-specific Markers in Acipenser sinensis Using AFLP

中文摘要：性别鉴定是生产养殖和物种保护中常用的技术。中华鲟 Acipenser sinensis是我国特有的一种大型海河洄游性鱼类,已极度濒危。由于中华鲟性成熟时间长,缺乏第二性征,基于外部特征难以进行性别鉴定,因此,筛选中华鲟性别特异性分子标记具有重要意义。利用扩增片段长度多态性(AFLP)分子标记,用10条 EcoR I-ANN引物和8条 Mse I-CNN引物组成的80对引物,对9尾雄性和15尾雌性中华鲟个体进行PCR扩增和毛细管电泳荧光分型,检测其基因组DNA多态性,寻找与其性别相关的分子标记。在100~500 bp范围,共获得864个位点,其中具有多态性DNA位点411条,多态性位点比例为48.58%,并未发现雌雄特异性位点。但发现33个位点在雌雄个体中的比例存在较大差异,聚类分析显示大部分雄鱼聚为一支,雌鱼聚为一支,从而推断这些位点可能与中华鲟性别具有密切相关性。该研究首次尝试在中华鲟基因组中寻找性别特异性的AFLP分子标记,尽管未找到特异性标记,但这些数据为进一步研究中华鲟性别相关基因和性别决定机制奠定了基础。
英文摘要：Gender identification is a common technique for breeding and species conservation. Chinese sturgeon ( Acipenser sinensis), a critically endangered large anadromous species, usually spawns in the Yangtze River of China. It takes a long time for A. sinensis to reach sexual maturity. It is impossible to distinguish gender by morphological characters without external sexual dimorphism; therefore, it is important to screen sex-specific molecular markers in A. sinensis. In this study, the polymorphism of 9 male and 15 female Chinese sturgeon genome DNA were detected by amplified fragment length polymorphism (AFLP) with 80 primer pairs including ten EcoR I-ANN and eight Mse I-CNN. A total of 864 loci including 411 polymorphic loci (48.58%) ranging in size from 100 bp to 500 bp were obtained, however, no sex-specific markers were observed. Additionally, 33 loci showed different distributions between female and male groups using cluster analysis, and these loci might be closely related with A. sinensis gender identification. This investigation may provide basic data for further research on gender related genes and gender determination in A. sinensis. 2015,(): 714-718 收稿日期：2014-12-18 DOI：10.11984/j.issn.1000-7083.20140563 分类号：Q959.4 基金项目：中国三峡集团公司内部研究项目(中华鲟子一代养殖群体遗传谱系研究, 编号:Z11005) 作者简介：刘雪清(1985-),男,研究方向:鱼类遗传保护,E-mail:Liu_xt2001@163.com *通讯作者：杜合军,E-mail:duhejun21@126.com 参考文献： 郭柏福, 常剑波, 肖慧, 等. 2011. 中华鲟初次全人工繁殖的特性研究[J]. 水生生物学报, 35(6):940-945. 季维智, 宿兵. 1999. 遗传多样性研究的原理与方法[M]. 杭州:浙江科学技术出版社. 匡友谊, 佟广香, 尹家胜, 等. 2007. 呼玛河哲罗鱼遗传多样性的AFLP分析[J]. 中国水产科学, 14(4):615-621. 李静, 陈松林, 邓思平, 等. 2007. 半滑舌鳎雌性特异扩增片段长度多态性标记的筛选与应用[J]. 水产学报, 31(5):591-597. 李静, 陈松林, 温海深. 2006. 鱼类性别相关基因及性别特异标记的研究进展[J]. 海洋水产研究, 27(4):90-95. 四川省长江水产资源调查组. 1988. 长江鲟鱼类生物学及人工繁殖研究[M]. 成都:四川科学技术出版社. 易继舫, 刘灯红, 唐大明, 等. 1999. 蓄养中华鲟的性腺发育与人工繁殖初报[J]. 水生生物学报, 23(1):85-86. Cui JZ, Shen XY, Gang QL, et al. 2006. Identification of sex markers by cDNA-AFLP in Takifugu rubripes[J]. Aquaculture, 257(1):30-36. Devlin RH, Nagahama Y. 2002. Sex determination and sex differentiation in fish:an overview of genetic, physiological, and environmental