叶绿体微卫星分析濒危植物资源冷杉的遗传多样性

Study on Genetic Diversity of the Endangered Plant Abies ziyuanensis by Using Chloroplast SSR Marker

用叶绿体微卫星(cpSSR)分子标记研究资源冷杉(Abies z iyuanensis)的遗传变异。资源冷杉分别取样于广西资源县和湖南城步县交界的银竹老山、湖南东安和新宁交界处的舜黄山、湖南炎陵县大院的7个居群。样品采用CTAB法进行总DNA的提取,用6对cpSSR引物进行扩增,扩增产物经95℃变性后在6%的变性聚丙烯酰胺凝胶上进行电泳,然后应用POPGEN V ersion 1.31软件计算资源冷杉各个居群的N ei's基因多样性指数(H e),Shannon信息多样性指数(I),基因分化系数(G st)以及N ei's遗传距离。用NTSY Spc V ersion 2.10软件进行U PGMA聚类分析,并对地理距离和遗传距离相关性进行M en tal检验。用ARLEQU IN 2.000软件分析地区间、居群间和居群内的遗传变异,统计有效单型模式数量。结果6对cpSSR引物中选出3对多态性引物,共扩增出7个位点。N ei's基因多样度H e为0.2922,信息多样性指数I为0.4428,基因分化系数G st为0.5805。资源冷杉的遗传分化主要存在于居群间,分布在银竹老山和舜黄山的资源冷杉与分布在大院的资源冷杉之间产生了明显的遗传分化,两地资源冷杉之间的遗传变异占总的68.87%。

The genetic variation of Abies ziyuanensis was studied by using chloroplast simple-sequence repeats （cpSSR）markers. The individuals of seven populations of A. ziyuanensis were sampled from Yinzhulaoshan of Guangxi,Shunhuangshan of Hunan and Dayuan of Hunan. Total genomic DNA was extracted using CTAB protocol. The amplification reaction was performed with six pair primers,and the fragments were denaturalized with 95℃ before being loaded and run on a 6% denaturing polyacrylamide gel. The Nei＇s gene diversity （He）, Shannon information index （I）, the coefficient gene differentiation （Gst） and Nei＇s genetic distance were computed by POPGEN Versin 1.31. UPGMA dengrogram was constructed by NTSYSpc Version 2.10. The correlations between genetic distance and geographical distance were investigated using a Mantel test. Analysis of molecular variance was conducted to calculate variance components for variation between two regions,among populations, and within populations using ARLEQUIN version 2. 000. Haplotypes were analyzed using this software. Seven loci were amplified by three pair primers screened from six pair primers. Nei＇s gene diversity （He） was 0. 2922, Shannon information index （I） was 0. 4428 and the coefficient gene differentiation （Gst） was 0. 5805. Genetic variation of Abies ziyuanensis were mainly existed among populations. High level of genetic differentiation was detected between the populations in Yinzhulaoshan and Shunhuangshan and in Dayuan,and the genetic variation between the two regions totally accounted for 68. 87 %.