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应用生态学报 ›› 2004, Vol. ›› Issue (2): 186-190.

• 研究论文 • 上一篇    下一篇

高黎贡山旱冬瓜Frankia的IGS PCR-RFLP分析

代玉梅, 曹军, 唐晓萌, 张成刚   

  1. 中国科学院沈阳应用生态研究所, 沈阳, 110016
  • 收稿日期:2003-06-18 修回日期:2003-08-18 出版日期:2004-02-15
  • 通讯作者: 张成刚
  • 基金资助:
    国家重点基础研究发展规划资助项目(001CB1089)

Diversity of Frankia in nodules of Alnus nepalensis at Gaoligong mountains revealed by IGS PCR-RFLP analysis

DAI Yumei, CAO Jun, TANG Xiaomeng, ZHANG Chenggang   

  1. Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China
  • Received:2003-06-18 Revised:2003-08-18 Online:2004-02-15

摘要: 在云南省高黎贡山自然保护区海拔1310~2400m的范围内,采集30个旱冬瓜根瘤样品,直接从根瘤中提取Frankia DNA,对其nifD-nifK基因间隔区(intergenic spacer,IGS)和16S-23S rDNA IGS进行PCR-RFLP分析.结果表明,nifD-nifK IGS的PCR产物长度差异很大,经HaeⅢ和AfaⅠ双酶切后,得到15种酶切带型,检测到多种基因型的菌株同时与同一株宿主植物共生;16S-23SrDNA IGS的PCR产物长度相似,酶切后亦区分出15种酶切带型.通过对两个基因间隔区的PCR-RFLP联合分析,发现高黎贡山旱冬瓜Frankia存在20种基因型.

Abstract: nodule samples were used to assess the diversity of Frankia strains symbiotically associated with Alnus nepalensis naturally occurring at the Gaoligong Mountains in Yunnan Province, China. DNA was extracted directly from nodules, and its two target DNA regions that encode nifD-nifK intergenic spacer (IGS) and 16S-23S rDNA IGS was studied by PCR-RFLP. The PCR fragments yielded by the nifD-nifK IGS were noticeably different in size, and when they were digested by HaeⅢ and AfaⅠ, 15 nif-type Frankia strains could be detected, the PCR-RFLP result of this region also could show that more than one genotype Frankia strains could form symbiosis with individual plants at the same time.The 16S-23S rDNA IGS had similar PCR fragments, but still identified 15 rrn-type strains after digested by HaeⅢ and AfaⅠ. 20 genotype strains could be found only when combined the PCR-RFLPs of two target regions.

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