悬钩子属植物rDNA ITS序列的克隆与分析

Cloning and analysis of rDNA ITS sequences from plants in Rubus L.

目的通过测定和分析15种悬钩子属Rubus L.药用植物的rDNA ITS序列,为分子鉴定和遗传多样性研究奠定基础。方法以叶片基因组DNA和通用引物为材料,用PCR法克隆rDNAITS全长,并利用生物信息学软件对序列进行分析。结果 15种悬钩子属植物ITS1、ITS2和5.8 S的序列长度分别为255～258、208～211和164 bp;ITS1和ITS2序列有变异位点138个,其中信息位点41个,序列存在较多的颠换、转换和缺失现象;5.8 S序列较为保守,仅含4个变异位点,没有发现信息位点;15种悬钩子属植物的遗传距离为0.139 0～0.008 1,灰毛泡和锈毛莓的遗传距离最小。结论获得了15种悬钩子属药用植物的rDNAITS序列,为分子鉴定和遗传多样性研究奠定了基础。

Objective To provide the evidences for molecular identification as well as genetic diversity studies, sequencing and comparison ofrDNA ITS sequences from 15 plants in Rubus L. were performed. Methods Full length rDNA ITS sequences were isolated from leaf genomic DNA by PCR method with universal primers, and these sequences was analyzed using bioinformatic soft-wares. Results The length of ITS1, ITS2, and 5.8 S sequences for the 15 plants inRubus L. were 255--258, 208--211, and 164 bp, respectively. Total 138 variable sites were found in ITS1 and ITS2 sequences with 41 parsimony information ones, and some transitions, transversions, and deletions were observed. The 5.8 S sequences were more conserved and there were only four variable sites with no parsimony information ones. The genetic distance of the 15 plants in Rubus L. ranged from 0.139 0 to 0.008 1, and the smallest genetic distance was observed between R. irenaeus and R. reflexus. Conclusion The rDNA ITS sequences of the 15 plants in Rubus L. are obtained, which would provide the evidences for molecular identification and genetic diversity studies.