基于ITS2序列的秤星树等冬青属8种药用植物的分子鉴别

目的]建立ITS2区段的秤星树及其同属几种药用植物的分子鉴别方法。方法]以秤星树等几种药用植物为材料,采用离心柱型试剂盒提取总DNA,采用一对通用引物对ITS2区段进行PCR扩增和测序;HMMer法对测序结果进行注释得到ITS2序列;MEGA软件分析,寻找差异位点,构建系统发生树。结果]测序后注释得到的秤星树等8种冬青属药用植物ITS2比对序列为254 bp,种间存在85个差异位点。结论]ITS2区可用于鉴别秤星树与同属几种药用植物。

Molecular Identification of Ilex asprella with Other Species in the genus Ilex L. Based on ITS2 Sequence Analysis

Objective] To establish molecular identification method for Ilex asprella and relative medicinal plants in the genus Ilex base on ITS2 sequence.Method] After DNA was extracted by reagent kit(centrifugal columnar),a pair of universal primers were used for PCR amplification and sequencing of ITS2 region.Then,the sequences of different samples were annotated by HMMer method to acquire ITS2 sequences.Based upon the multiple sequence alignment of the amplified ITS2 sequences,a phylogenetic tree was constructed using the software MEGA.Result] 254 bp ITS2 fragment of each species were obtained,and there were 85 different base sites between species.Conclusion] ITS2 region can be used to identify Ilex asprella from relative medicinal plants in the genus Ilex L..