采用ISSR-PCR技术对曼氏无针乌贼基因组DNA进行扩增，筛选分析了ISSR—PCR扩增时的主要影响因子，包括Mg^2＋浓度、引物浓度、dNTPs浓度、Taq酶浓度以及退火温度，并用8条ISSR引物对舟山养殖群体进行了遗传多样性分析，最终确立的适用于曼氏无针乌贼ISSR扩增的25μl反应体系为：Mg2＋1．5mmol／L，TaqDNA聚合酶1．0U，引物0．15μmol／L，dNTPs2．0mmol／L，退火温度为52．2℃。结果表明，扩增得到70个清晰的扩增位点，其中多态性位点60个，多态位点率为85．71％，Nei’S基因多样性为0．3105±0．1760，Shannon’S多样性指数为0．4610±0．2397。本研究结果表明，舟山曼氏无针乌贼养殖群体遗传多样性较丰富，因此舟山曼氏无针乌贼养殖群体可能有比较高的适应生存的能力，蕴藏着比较大的进化潜能及比较丰富的育种潜力。%In this study, ISSR-PCR amplification technology was adopt on Sepiella maindroni genome DNA, and the effect of Mg2＋ concentration, Taq DNA polymerase, dNTPs, primer and the annealing temperature on the ISSR-PCR amplification was analyzed. 8 ISSR primers were used to assess the genetic diversity of Zhoushan cultured S. maindroni, and established the suitable ISSR-PCR system with a total volume of 25μl was as followed： Mgz＋ 1.5mmol/L, Taq DNA polymerase 1.0U, primer 0.15μmol/L, dNTPs 2.0mmol/L, and the annealing temperature is 52.2℃. Of the 70 ISSR loci tested, 60 （85.71%） were polymorphic, Nei＇s genetic diversity is 0.3105±0.1760, Shannon＇s information index is 0.4610±0.2397. The results of this study showed that there was a higher genetic diversity level in the Zhoushan cultured population of S. maindroni. Therefore, Zhoushan cultured S. maindroni maybe have higher adapt ability to survive, containing larger evolution potential and relatively rich breeding potential.
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