摘要
通过分析青天葵及其常见混伪品的ITS2序列,建立青天葵新型真伪鉴别方法。采用植物基因组DNA提取试剂盒提取青天葵及其混伪品的叶片DNA,一对通用引物PCR扩增ITS2基因片段并直接双向测序。采用DNAMAN、ClustalX软件拼接比对序列,MEGA4.0软件构建NJ树,Schultz等建立的数据库和网站预测lTS2序列的二级结构。结果显示,获得的12条ITS2序列的长度范围为203-242 bp,GC含量范围为53.1%-71.8%。所有样品ITS2序列比对后的长度为249 bp,其中存在226个变异位点。青天葵种间K2P遗传距离(1.125)远大于种内K2P遗传距离(0.004)。基于ITS2的序列的NJ树和二级结构均能直观地区分青天葵及其混伪品。
It was to establish a novel identification method for Nerviliafordii and its aduherants through ITS2 sequence analysis. Total I）NA was extracted from fresh leaves, and ITS2 region was amplified and sequenced with a pair of universal primers. The sequences were assembled and aligned by DNAMAN and CLUSTALX, and NJ tree was constracted using MEGA 4.0. Secondary structures of ITS2 sequences were predicated in the database built by Schuhz et al. Significant differences were observed in the ITS2 sequences between Nerviliafordii and its adulterants. The interspecific genetic distance was far more than the intraspecific disatance. Both NJ tree and secondary structure based on ITS2 sequences can distinguish Nerviliafordii and its adulterants intuitively. ITS2 is an effective biomarker to identify Nerviliafordii and its adulterants.
出处
《生物技术通报》
CAS
CSCD
北大核心
2012年第12期173-179,共7页
Biotechnology Bulletin
基金
教育部博士点基金联合资助项目(200805720004)
教育部留学回国人员科研项目[教外司留(2009)1001]
关键词
青天葵
ITS2混伪品
分子鉴别
Nerviliafordii （ Hance ） Schltr. ITS2 adulterant Molecular identification
