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本研究采用改良CTAB法、改良SDS法、高盐低p H法提取自然风干的水母雪莲植株的基因组DNA,并对提取方法进行优化筛选出适合于水母雪莲药材DNA的提取方法。研究表明改良CTAB(CTAB浓度为2%,PVP浓度为1%,β-巯基乙醇浓度为2%)法所提取的基因组DNA得率较高,且使用氯仿/异戊醇抽提两次时提取出的DNA质量较好,能够满足ISSR分析要求。在此基础上对实验材料进行预处理,优化获得了完全适用于水母雪莲风干药材DNA提取的一整套提取流程。该方法能够有效去除次生代谢产物对DNA提取的影响,且对原材料消耗较少,可以应用于后续水母雪莲种质资源鉴定和遗传多样性分析。
In this study, genomic DNA was extracted from naturally dried Saussurea elaphora using improved CTAB method, modified SDS method and high salt and low p H method, and the extraction method was optimized for DNA extraction from medicinal materials of Saussurea medusa. Studies have shown that genomic DNA extracted from modified CTAB (2% CTAB, 1% PVP, 2% β-mercaptoethanol) yields a higher yield of genomic DNA extracted twice with chloroform / isoamyl alcohol The quality of the DNA is good enough to meet the ISSR analysis requirements. On this basis, the experimental materials were pretreated, and the complete extraction procedure was completely optimized for the DNA extraction of dried jellyfish Saussurea officinalis. The method can effectively remove the effects of secondary metabolites on DNA extraction and consume less raw materials, and can be applied to the identification of germplasm resources and the analysis of genetic diversity in subsequent seasons.