高黎贡山旱冬瓜Frankia的IGS PCR-RFLP分析  

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

作　　者：代玉梅[1] 曹军[1] 唐晓萌[1] 张成刚[1] 

机构地区：[1]中国科学院沈阳应用生态研究所,沈阳110016

出　　处：《应用生态学报》2004年第2期186-190,共5页Chinese Journal of Applied Ecology

基　　金：国家重点基础研究发展规划资助项目 ( 0 0 1CB10 89)

摘　　要：在云南省高黎贡山自然保护区海拔 1310～ 2 4 0 0m的范围内 ,采集 30个旱冬瓜根瘤样品 ,直接从根瘤中提取FrankiaDNA ,对其nifD nifK基因间隔区 (intergenicspacer,IGS)和 16S 2 3SrDNAIGS进行PCR RFLP分析 .结果表明 ,nifD nifKIGS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 2 3SrDNAIGS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 2nodule 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.

关 键 词：Frankia基因型 高黎贡山 IGS PCR—RFLP 

分 类 号：S792[农业科学—林木遗传育种]