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›› 2020, Vol. 42 ›› Issue (4): 324-338.

• 论文与报告 • 上一篇    下一篇

双线紫蛤COI基因序列多态性与水环境分析

王朋云   

  1. 莆田市水产技术推广站
  • 收稿日期:2020-03-19 修回日期:2020-04-25 出版日期:2020-08-25 发布日期:2020-08-18
  • 通讯作者: 王朋云
  • 基金资助:
    福建省海洋生物增养殖与高值化利用重点实验室开放课题;国家贝类产业技术体系福建综合试验站项目;福建省渔业“五新”项目

Analysis of COI genetic sequence polymorphism and habitat water environment of Soletellina diphos

Peng-Yun WANG   

  • Received:2020-03-19 Revised:2020-04-25 Online:2020-08-25 Published:2020-08-18
  • Contact: Peng-Yun WANG

摘要: 以双线紫蛤(Soletellina diphos)为研究对象,测定南日岛海区自然群体30枚个体的线粒体COI基因部分序列,研究其个体间的序列单核苷酸多态性(SNP),掌握双线紫蛤的遗传多样性;统计南日岛海域2008年4月—2018年12月期间海水样品的盐度、温度、pH、溶解氧、叶绿素a、透明度、化学需氧量、无机氮、无机磷、石油类10项理化因子检测结果,分析数据随季节月份变化状况,了解双线紫蛤栖息地水环境。结果显示,南日岛双线紫蛤群体30枚个体的序列中碱基T、C、A、G平均含量分别为41.95%、16.07%、18.53%、23.45%,(A+T)含量明显高于(G+C)含量,共发现13个核苷酸多态位点(转换/颠换偏倚率13.04),具有9个单倍型,有1个为引起亮氨酸/苯丙氨酸变化的非同义突变位点,单倍型多样性指数(Hd)为(0.598 0±0.103 0),核苷酸多样性(Pi)为(0.002 9±0.000 6),平均核苷酸变异数(k)为1.912 6,表现出略高于标准值的单倍型多样性和较低的核苷酸多样性,个体间的遗传距离为0~0.009 2、平均值为(0.002 9±0.001 0),Tajima’s D和Fu’s Fs中性检验的结果均为不显著负值,说明这个群体的双线紫蛤遗传相对稳定。南日岛产区水环境月均盐度变化范围为26.84~34.33、温度变化范围为13.27~29.33℃、pH变化范围为7.89~8.54、溶解氧变化范围为5.64 ~9.31 mg/L、叶绿素a变化范围为0.33 ~15.57 μg/L、透明度变化范围为0.40 ~1.79 m、化学需氧量变化范围为0.22 ~1.59 mg/L、无机氮变化范围为0.04 ~0.45 mg/L、无机磷变化范围为0~0.16 mg/L、石油类变化范围为0 ~0.20 mg/L,盐度、温度在各年春、夏、秋三季变化皆先升后降,夏季最高;pH、溶解氧则在春、夏两季波动呈下降;而无机氮在春、夏、秋季趋势为先降后升,夏季最低;除了化学需氧量和石油类以外,其余参数的周年变动均表现出一定的季节性差异规律,同时表明双线紫蛤在以上的理化条件下都可以进行生长繁殖。

关键词: 双线紫蛤, COI基因, 序列多态性, 栖息地水环境, Soletellina diphos, COI gene, sequence polymorphism, habitat water environment

Abstract: Partial mitochondrial COI gene region of 30 individuals of Soletellina diphos collected from Nanri Island, Fujian. China, were cloned by PCR and directly sequenced from both directions to detect the Single Nucleotide Polymorphism (SNP) among individuals and genetic diversity of the natural population on molecular level. Evidently, the content of (A+T) base in COI was higer than that of (G+C) base, and its average nucleotide percentage composition of T, C, A and G were 41.95%, 16.07%, 18.53% and 23.45%, respectively. According to the mutation form, mutation frequency and mutation distribution, a total of 13 SNP loci were detected in those 655 base pairs of gene fragments, containing 12 transformations and 1 transversion (R=13.04). And 9 haplotypes were found in 30 samples through multi-sequences alignment. One SNP locus located at 203 bp of which was nonsynonymous mutation that caused the change between Leucine and Phenylalanine, while the other 12 variation sites had synonymous mutations after amino acid analysis. There was no distinct relevancy between different SNP loci. Population of S. diphos displayed just slightly higher than standard haplotype diversity index (Hd) of (0.598 0 ± 0.103 0) and relatively lower nucleotide diversity index (Pi) of (0.002 9 ± 0.000 6), with the average nucleotide difference (k) of 1.912 6 based on COI sequences. The Kimura 2-parameter genetic distance between different individuals in the specimen ranged from 0 to 0.009 2, and the total average distance was (0.002 9 ± 0.001 0). Results of both the Tajima’s D and Fu’s Fs neutral tests were not significantly negative, illustrating that the inheritance of this population was relatively stable. Historical test results of seawater samples collected around the area of Nanri Island from April 2008 to December 2018 were detailed analyzed in order to get to know about the habitat conditions of S. diphos through obtaining the seasonal and monthly changing rule of 10 physical and chemical indexes such as salinity, temperature, pH, dissolved oxygen, chlorophyll a, transparency, chemical oxygen demand, inorganic nitrogen, inorganic phosphorus and petroleum. The annual trend of water environment factors data expressed that S. diphos mainly lived in the intertidal zone and shallow water where the range of monthly average seawater salinity, temperature, pH, dissolved oxygen, chlorophyll a, transparency, chemical oxygen demand, inorganic nitrogen, inorganic phosphorus and petroleum was 26.84~34.33, 13.27~29.33℃, 7.89~8.54, 5.64~9.31 mg/L, 0.33 ~15.57 μg/L, 0.40 ~1.79 m, 0.22 ~1.59 mg/L, 0.04 ~0.45 mg/L, 0~0.16 mg/L and 0 ~0.20 mg/L, respectively. The yearly changes of salinity and temperature from spring to autumn rose and then fell, with the highest value in summer. Every spring to summer, pH and dissolved oxygen demonstrated a fluctuation of decrease in the same way. Meanwhile the tendency of inorganic nitrogen decreased first and increased later from spring to autumn, with the lowest value in summer. Except for chemical oxygen demand and petroleum content, the annual changes of the other three factors all showed an obvious or a certain seasonal difference rule. It also indicated that S. diphos could grow and reproduce well under the above physical and chemical conditions of seawater environment in the nearshore area of Nanri Island.

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