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| 香鱼(Plecoglossus altivelis)养殖群体遗传多样性的AFLP分析及性别特异性分子标记筛选 |
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闫松松,苗亮,李明云①,范慧慧,胡谋,陈炯,史雨红
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宁波大学 应用海洋生物技术教育部重点实验室,宁波大学 应用海洋生物技术教育部重点实验室,宁波大学 应用海洋生物技术教育部重点实验室,宁波大学 应用海洋生物技术教育部重点实验室,宁波大学 应用海洋生物技术教育部重点实验室,宁波大学 应用海洋生物技术教育部重点实验室,宁波大学 应用海洋生物技术教育部重点实验室
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| 摘要: |
| 采用扩增片段长度多态性(AFLP)技术分析了香鱼(Plecoglossus altivelis)养殖群体的遗传多样性, 并筛选性别特异性分子标记。结果表明, 15 对选扩引物组合共扩增到889个位点, 其中多态性位点380个, 多态率为42.74%; 群体Nei氏遗传多样性指数为0.1454, Shannon氏指数I为0.2174。香鱼养殖群体的遗传多样性较丰富, 处于中等偏上水平。仅引物组合E-ATG/M-CTG扩增到1条雄性特异性条带, 长度为139bp。以该雄性特异性AFLP条带的DNA序列为模板, 设计1对特异的PCR引物并在10尾已知性别的香鱼(雌雄各5尾)中扩增检验, 结果显示5尾雄鱼均可扩增到目的条带而5尾雌鱼均无扩增。表明该序列是雄性特异性分子标记, 可用于鉴别香鱼的遗传性别, 并为香鱼的性别决定机制和性别控制研究奠定基础。 |
| 关键词: 香鱼 扩增片段长度多态性(AFLP) 养殖群体 遗传多样性 性别特异性标记 |
| DOI：10.11693/hyhz20140100006 |
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| 基金项目:浙江省科技厅重大科技专项, 2009C12077 号; 国家“973”计划前期研究专项, 2008CB117015 号; 长江学者和创新团队发展计划项目, IRT0734 号。 |
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| MARKER BY AFLP IN CULTURED PLECOGLOSSUS ALTIVELISPOPULATION |
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yansongsong,miaoliang,limingyun,fanhuihui,humou,chenjiong and shiyuhong
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Key Laboratory of Applied Marine Biotechnology, Ministry of Education Ningbo University,Key Laboratory of Applied Marine Biotechnology, Ministry of Education Ningbo University,Key Laboratory of Applied Marine Biotechnology, Ministry of Education Ningbo University,Key Laboratory of Applied Marine Biotechnology, Ministry of Education Ningbo University,Key Laboratory of Applied Marine Biotechnology, Ministry of Education Ningbo University,Key Laboratory of Applied Marine Biotechnology, Ministry of Education Ningbo University,Key Laboratory of Applied Marine Biotechnology, Ministry of Education Ningbo University
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| Abstract: |
| We analyzed the genetic diversity of cultured Plecoglossus altivelispopulation to isolate sex-specific marker in amplified fragment length polymorphic (AFLP) technique. Forty sex-matured fish individuals (70—120g) in two batches were randomly sampled from a localfish farm. The first batch included 30 (15 each for male and female) in Sep. 2010 for AFLP analysis; and the second batch included 10 (5 each for male and female) in Sep. 2011 for sex-specific marker isolation. A total of 889 bands were obtained using 15 pairs of AFLP primer, of which 380 loci were polymorphism taking 42.74% of the total. The Nei’s gene diversity and Shannon index was 0.1454 and 0.2174, respectively. The results
indicate that the cultured P. altivelispopulation was in medium genetic diversity. Among all, only a primer pair E-ATG/M-CTG amplified a band that presented in all males but females. The length of this male-specific AFLP marker was 139bp. Specific PCR primers were then designed and PCR amplification performed on the second batch 10 sex-known individuals of P. altivelis. Results show that the objective band could be amplified in all males but females. We believe that that the marker is male-specific and can be used to identify the genetic sex of P. altivelis. This study may contribute to the sex-specific molecular marking and to sex determination and control in P. altivelis. |
| Key words: Plecoglossus altivelis AFLP cultured population gene diversity sex-specific molecular marker |