中文摘要：

目的：建立适合新疆贝母属植物总DNA的ISSR-PCR优化反应体系,为新疆贝母属8种药用贝母品种鉴定和遗传多样性研究提供依据。方法：综合利用单因素实验和L16（45）正交试验,对因素Taq DNA聚合酶、d NTPs、Mg2＋、引物浓度、模板DNA含量进行优化,并考察循环次数、延长时间及退火温度。结果：每个因素的不同水平对PCR反应有显著影响,其中Mg2＋影响最大。新疆贝母属植物总DNA的ISSR-PCR最适体系为50μL：5μL 10×Buffer、2.5 mmol/L Mg2＋、0.30 mmol/L d NTPs、0.8μmol/L引物、0.75 U Taq酶、1.0 ng/μL模板DNA、33.35μL dd H2O。循环次数为45 cycle,延伸时间10 min,引物UBC853的最适退火温度为49.8℃。结论：建立的新疆贝母属植物总DNA的ISSR-PCR反应体系经23份新疆贝母属8种药用贝母样品检验,ISSR-PCR扩增效果显著,证明该体系具有较高的稳定性和可重现性,为新疆贝母属8种药用贝母遗传多样性的分子标记研究奠定基础。

英文摘要：

Objective： To optimize the ISSR-PCR reaction system of 8 species of plants in FritiUaria L. from Xinjiang which will provide a scientific basis for identification and genetic diversity analysis. Meth- ods ： Comprehensive utilization of single factor experiment and L16 （4^5 ） orthogonal test, the factors of Taq DNA polymerase, dNTPs, Mg^2＋ , concentration of primers, template DNA content is optimized, and the effects of cycle number, extension of time and annealing temperature. Results： Every factor in different levels had the significant effects on the result of PCR, and the most remarkable factor was the concentra- tion of Mg^2＋. ISSR-PCR optimal system for total DNA of plants in Fritillaria L. from Xinjiang as the 50 μl：5 μl 10 × Buffer, 2.5 mmol/L Mg^2＋ , 0.30 mmol/L dNTPs, 0.8 μmol/L primer, 0.75 U Taq DNA polymerase, 1.0 ng/μl template DNA, 33.35 μl ddH20. The cycle times of 45 cycle, extended time of 10 min, the suitable annealing temperature of primer UBC853 was 49.8 ℃. Conclusion ： The established and optimized ISSR reaction system is stable and credible and ISSR-PCR amplification effect is obvious according to the testing results of 23 samples of 8 species of plants in Fritillaria L. from Xinjiang, and lay the foundation for the study of molecular markers of genetic diversity in 8 species of plants in Fritillaria L. from Xinjiang.