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适于AFLP分析用的桃成熟叶片DNA提取方法

DNA extraction method for AFLP analysis in mature leaves of peach

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【作者】 王富荣赵剑波章镇佟兆国姜立杰姜全张开春

【Author】 WANG Fu-rong1,2,ZHAO Jian-bo2,ZHANG Zhen1*,TONG Zhao-guo1,JIANG Li-jie2,JIANG Quan2,ZHANG Kai-chun2 (1College of Horticulture,Nanjing Agricultural University,Nanjing,Jiangsu 210095 China;2Institute of Forestry and Pomology,Beijing Academy of Agricultural and Forestry Sciences,Beijing 100093 China)

【机构】 南京农业大学园艺学院北京市农林科学院林业果树研究所 北京100093北京市农林科学院林业果树研究所南京210095北京100093

【摘要】 以多个野生桃秋季成熟叶片为材料,采用改进CTAB法对其基因组总DNA的提取进行了研究,并与其相应幼叶DNA的提取效果进行了比较分析。结果表明,从野生桃成熟叶片提取的DNA除产量低于幼叶外,DNA的纯度和完整性都较好,D260nm/D280nm的比值均为1.8~2.0,无降解现象,RNA去除干净,能被限制性内切酶完全消化;其AFLP分析(引物EcoRI-TA/MseI-CTT)条带清晰,多态性好,说明此方法提取的野生桃成熟叶片基因组总DNA完全适于AFLP分析。

【Abstract】 The genomic DNA samples of mature leaves of several wild peach varieties in autumn were isolated by improved CTAB method,and DNA extraction effects were contrasted with their tender leaves. The results showed: the extracted DNA of mature leaves by this method was pure,integral,the value of D260 nm/D280 nm was 1.80 to 2.0,degradation was nearly free,the RNA was eliminated completely,and suitable for digestion by restriction endonucleases,except the yields of DNA extraction from mature leaves was lower than that of the tender leaves. AFLP analysis indicated the primer (EcoRI-TA/MseI-CTT) produced clear polymorphic patterns. Therefore this method could be used for extracting ideal DNA samples from mature leaves of wild peach and suitable for AFLP.

【关键词】 DNA提取成熟叶片AFLP
【Key words】 PeachDNA extractionMature leavesAFLP
【基金】 北京市自然科学基金资助项目(5042007)
  • 【文献出处】 果树学报 ,Journal of Fruit Science , 编辑部邮箱 ,2006年04期
  • 【分类号】S662.1
  • 【被引频次】28
  • 【下载频次】306
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