不同产地菊苣的ITS序列分析及模式识别研究

目的 通过分析不同产地菊苣Cichorium intybus和毛菊苣C.glandulosum核糖体基因内部转录间隔区(ribosomal DNA internal transcribed spacer,rDNA-ITS)序列,为菊苣药材鉴定和品种鉴别提供DNA分子标记.方法 采用广谱植物基因组DNA快速提取试剂盒提取14个产地的菊苣和毛菊苣样本的总DNA,利用ITS序列扩增通用引物PCR扩增rDNA-ITS序列并测序.运用DNAMAN V6软件比较两品种菊苣样品ITS序列的差异.所有样品ITS序列上的差异碱基经数学处理后使用SPSS 17.0软件对其进行聚类分析.结果 菊苣ITS序列种内相似度基本上在99.2％以上,毛菊苣ITS序列种内相似度在99.8％以上,而两品种间ITS序列相似度在99.2％以下.两品种菊苣的ITS序列上分别有多个特异性信息位点.聚类分析实现了两品种菊苣ITS序列差异的识别.结论 rDNA-ITS序列是菊苣和毛菊苣药材鉴定和品种鉴别的有效分子标记.

ITS sequence analysis and gene pattern recognition of Cichorium intybus from different habitats

Objective To analyze the ribosomal DNA internal transcribed spacer (rDNA-ITS) sequence of Cichorium intybus and C. glandulosum from different habitats, and to provide DNA molecular marker for the identification of chicory. Methods The total DNA was extracted from the samples of C. intybus and C. glandulosum from 14 habitats by rapid broad spectrum of plant genomic DNA extraction kit. The ITS sequence was amplified by PCR with universal primer of ITS and then sequenced. The two kinds of ITS sequences were compared by DNAMAN V6 software. The cluster analysis was adopted by SPSS 17.0 after the different ITS bases from all the samples were mathematically treated. Results The intraspecies identity of ITS sequence was above 99.2% in C. intybus, and that in C. glandulosum was above 99.8%, while the interspecies identity of ITS sequence was below 99.2%. There were various specific information sites in the ITS sequences of the two kinds of chicory. Conclusion The ITS sequence is an available molecular marker for the identification of C. glandulosum and C. intybus.