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目的通过测定和分析11种堇菜属药用植物的核糖体DNA内转录间隔区(r DNA ITS)序列,为该属药用植物的分子鉴定提供依据。方法利用PCR法克隆ITS序列,借助生物信息学工具进行序列分析、遗传距离估算和系统发育树构建。结果 11种堇菜属植物的ITS全长为612~638 bp,ITS1与ITS2的长度为251~265 bp和198~211 bp,5.8 S的长度十分保守,均为163 bp;ITS1、ITS2与5.8 S的信息位点数分别为50、23和3。11种堇菜属植物的遗传距离为0.025~0.137,紫花堇菜与七星莲的遗传距离最大,亲缘关系最远;紫花堇菜和鸡腿堇菜的遗传距离最小,亲缘关系最近。结论克隆到堇菜属11种药用植物的ITS序列,它们的信息位点丰富,可用于分子鉴定。
OBJECTIVE: To provide a basis for the molecular identification of medicinal plants belonging to this genus by measuring and analyzing the sequence of ribosomal DNA transcribed spacer (rDNA ITS) in 11 species of Viola medicinal plants. Methods The ITS sequences were cloned by PCR and sequenced with bioinformatics tools. The genetic distances were estimated and phylogenetic trees were constructed. Results The ITS sequences of 11 species of Viola were 612-638 bp in length and 251-265 bp and 198-211 bp in ITS1 and ITS2, respectively. The length of 5.8 S was very conservative, both of which were 163 bp. ITS1, ITS2 and 5.8 S were 50, 23 and 3. The genetic distances among the 11 Viola species were 0.025-0.13 7. The genetic distance between Viola violacea and Qixinglian was the largest and the genetic distance was the furthest. Viola and Viola The genetic distance is the smallest, the closest relationship. Conclusion The ITS sequences of 11 species of Viola were cloned and their informative sites were rich and could be used for molecular identification.