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作 者:周俊[1] 张冰[1] 吴丽丽[2] 林志健[1] 朱文静[1] 孙博喻[1] 王红坡[1]
机构地区:[1]北京中医药大学中药学院,北京100102 [2]赣州市人民医院药剂科,江西赣州341000
出 处:《中草药》2013年第13期1823-1826,共4页Chinese Traditional and Herbal Drugs
基 金:国家自然科学基金资助项目(81073068;81274153);国家教育部博士点基金(20120013130002);北京中医药大学科研创新团队项目(2011-CXTD-14);国家级教学团队建设项目(07td010);北京市优秀教学团队建设项目(2007012)
摘 要: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(ribosomalDNA 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。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.
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