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皱边石杉内生真菌DNA提取有效方法比较

Compared Two Effective DNA Extraction Methods to Extract the Genome Total DNA from the Endophyte Fungus of Huperzia crispata

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【作者】 禹利君高思青史云峰杨培迪张慧黄歌阳

【Author】 YU Li-jun,GAO Si-qing,SHI Yun-feng,YANG Pei-di,ZHANG Hui,HUANG Ge-yang(College of Horticulture and Landscape in Hunan Agricultural University;Hunan Provincial Research Center of Natural Products,Changsha 410128,China)

【机构】 湖南农业大学园艺园林学院湖南省天然产物研究中心

【摘要】 目的:优化确定适用于皱边石杉内生真菌DNA提取的有效方法,并评价其效果。方法:运用氯化苄改良法、CTAB改良法分别提取皱边石杉内生真菌菌丝体DNA。结果:氯化苄改良法可从17个内生真菌菌丝体中均获得了高质量的DNA,除5、13、14号生长缓慢的内生真菌其菌丝体DNA浓度≤60ng/μL,其余样品的DNA浓度均≥200ng/μL。而CTAB改良法仅适宜于平板培养菌落生长迅速、菌落疏松,发酵培养菌丝体产量高的1、2、3、9、10、12号菌株。结论:氯化苄改良法是适宜于皱边石杉内生真菌DNA提取的理想方法。

【Abstract】 Objective: To optimize and evaluate the efficient DNA extraction method for endophyte fungus which were isolated from Huperzia crispate.Method: Using the improved benzyl chloride method and the improved CTAB method to extract DNA and compared their extraction efficiency and quality.Result: Using improved benzyl chloride method can obtain good quality DNA for 17 endophyte fungus mycelium.Except the DNA concentrations of No.5,13 and 14 were ≤60 ng/μL,the other samples DNA concentrations were ≥200 ng/μL.However,improved CTAB method was only suitable for No.1,2,3,9,10 and 12 fungus,which cloning was soft and grow quickly in plate culture,ferment mycelium yields was high.Conclusion: The improved benzyl chloride method was the ideally total genome DNA extraction method for endophyte fungus of Huperzia crispate.

【关键词】 皱边石杉内生真菌DNA提取
【Key words】 Huperzia crispataendophyte fungusDNAextraction
【基金】 湖南省自然科学基金项目(07JJ6060);省教育厅项目(06C409)资助
  • 【分类号】S567.19
  • 【被引频次】9
  • 【下载频次】491
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