Dai Yumei, Cao Jun, Tang Xiaomeng, Zhang Chenggang
Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, China.
Ying Yong Sheng Tai Xue Bao. 2004 Feb;15(2):186-90.
30 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 Hae III and Afa I, 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 Hae III and Afa I. 20 genotype strains could be found only when combined the PCR-RFLPs of two target regions.
采用30个根瘤样本评估与中国云南省高黎贡山自然生长的尼泊尔桤木共生的弗兰克氏菌菌株的多样性。直接从根瘤中提取DNA,并通过PCR-RFLP研究其编码nifD-nifK基因间隔区(IGS)和16S-23S rDNA IGS的两个目标DNA区域。nifD-nifK IGS产生的PCR片段大小明显不同,用Hae III和Afa I消化后,可检测到15种nif型弗兰克氏菌菌株,该区域的PCR-RFLP结果也表明,不止一种基因型的弗兰克氏菌菌株可同时与单株植物形成共生关系。16S-23S rDNA IGS具有相似的PCR片段,但用Hae III和Afa I消化后仍鉴定出15种rrn型菌株。只有将两个目标区域的PCR-RFLP结果结合起来,才能发现20种基因型菌株。