华北农学报

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云南岩白菜资源的DNA提取和ISSR引物筛选

王仕玉1, 郭凤根2, 刘芸君2, 刘云芬2, 杨生超2, 张应华1   

  1. 1. 云南农业大学园林园艺学院, 云南 昆明 650201;
    2. 云南农业大学农学与生物技术学院, 云南 昆明 650201
  • 收稿日期:2012-08-10 出版日期:2012-12-31
  • 通讯作者: 郭凤根(1964 - ), 男, 浙江平湖人, 教授, 博士, 博士生导师, 主要从事植物资源的评价与利用研究。
  • 作者简介:王仕玉(1965 - ), 女, 重庆合川人, 副教授, 博士, 主要从事园艺作物和药用植物资源研究。
  • 基金资助:
    云南省社会发展科技计划(2007C211M);云南省科技厅重点产业创新工程项目(2008IF004)资助

DNA Extraction and ISSR Primer Screening of Resources of Bergenia purpurascens in Yunnan

WANG Shi-yu1, GUO Feng-gen2, LIU Yun-jun2, LIU Yun-fen2, YANG Sheng-chao2, ZHANG Ying-hua1   

  1. 1. College of Horticulture and Landscape Archetecture,Yunnan Agricultural University,Kunming 650201,China;
    2. College of Agronomy and Biotechnology,Yunnan Agricultural University,Kunming 650201,China
  • Received:2012-08-10 Published:2012-12-31

摘要: 提取滇产岩白菜资源的基因组DNA并进行ISSR引物的筛选,为用ISSR标记研究云南岩白菜资源的遗传多样性奠定基础。DNA提取采用改良的CTAB法,质量检测采用琼脂糖凝胶电泳法和紫外分光光度计法,ISSR引物选用加拿大哥伦比亚大学开发的100条引物。结果表明,从18份云南岩白菜资源的幼叶中提取了基因组DNA,浓度在1 387.5~12 000 ng/μL之间,A260/A280的值在1.61~1.85之间,琼脂糖凝胶电泳检测呈一条带;从100种ISSR引物中筛选出了22种扩增结果好的引物。所提DNA质量较高,该提取方法适用于从酚类和蛋白质含量高的植物材料中提取DNA;22个ISSR引物可用于岩白菜资源的遗传多样性研究。

关键词: 岩白菜, 云南, DNA提取, ISSR引物筛选

Abstract: Bergenia purpurascens ( Hook. f. et Thoms. ) Eng1 . is an endangered medicinal species mainly distributed in Yunnan, Sichuan and Tibet. There are a lot of reports about its chemistry and pharmacology but no reports on its genetic diversity and conservation. To reveal the genetic diversity of B. purpurascens in Yunnan, 18 genome DNA samples of B. purpurascens collected in Yunnan province were extracted from young leaves by modifiedCTAB methods and the screening of primers for ISSR analysis were conducted. The concentrations of DNA samplesvaried from 1 387. 5 ng / μL to 12 000 ng / μL while the ratios of A260 /A280were between 1 . 61 and 1 . 85. There wasonly one band on the electrophoretogram for each DNA sample. These results confirmed that the qualities of extracted DNA samples were good and the modified CTAB methods described in this paper could be used for extractingDNAs from plant materials with abundant polyphenol and protein. Twenty-two ISSR primers which gave good amplification results were screened out from 100 primers and this provided a solid basis for exploring genetic diversity ofB. purpurascens in Yunnan by ISSR molecular marker.

Key words: Bergenia purpurascens (Hook. f. et Thoms. ) Eng1 . , Yunnan, DNA extraction, ISSR primer Screening

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引用本文

WANG Shi-yu, GUO Feng-gen, LIU Yun-jun, LIU Yun-fen, YANG Sheng-chao, ZHANG Ying-hua. DNA Extraction and ISSR Primer Screening of Resources of Bergenia purpurascens in Yunnan[J]. ACTA AGRICULTURAE BOREALI-SINICA, doi: 10.3969/j.issn.1000-7091.2012.z1.005.

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