基于SNP位点鉴定藏菖蒲及其近缘种  

Molecular Authentication of the Oriental Medicine Acorus calamus Based on SNP Markers

作　　者：王丽丽[1] 林余霖[1] 陈晓辰[1] 廖保生[1] 王晓玥[1] 金钺[1] 韩建萍[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193

出　　处：《中国现代中药》2014年第11期895-900,共6页Modern Chinese Medicine

基　　金：国家自然科学基金资助项目(81473303);863计划(2012AA021602):基于DNA条形码的珍稀药用;野生等资源快速检测技术及产品研发;太白贝母甾体生物碱生物合成途径的比较转录组学研究

摘　　要：目的:采用DNA条形码技术和SNPs技术,快速准确鉴定藏菖蒲及其近缘种、混伪品。方法:提取藏菖蒲及其近缘种的全基因组DNA,扩增内部第二转录间隔区(internal transcribed spacer 2,ITS2)并测序,测序结果用Codon Code Aligner 4.2.7拼接;基于隐马尔科夫模型(Hidden Markov Model,HMM)注释ITS2,采用MEGA5.1进行序列比对,SNP位点查找、种内主导变异分析和系统进化树构建。结果:通过对藏菖蒲同属近缘种的96条ITS2序列进行分析,发现第23位和212位存在稳定的SNP位点,可准确鉴定藏菖蒲;藏菖蒲ITS2序列的种内变异很小,仅在158位存在一个多拷贝位点。结论 :该方法快速、准确、特异性强,可用于藏菖蒲的鉴定。Objective： The aim of this study is to investigate a simple and reliable method to authenticate Acorus calamus according to DNA barcoding and SNPs. Methods： In this study we sequenced 47 internal transcribed spacer 2 （ ITS2 ） regions of A. calamus and A. tatarinowii, was assembled by CodonCode Aligner 4. 2. 7 and annotated in accordance with HMM（ Hidden Markov Model）. MEGAS. 1 was used to analyze all the 104 ITS2 sequences and detect single nucleotide polymorphisms （SNPs）. Meanwhile, according to sequence diversity, a molecular phylogeny was constructed using neighbor-joining method. Intra-genomie variation of A. calamus and A. tatarinowii were investigated and compared with sequences obtained via direct PCR sequencing. Results： Two stable single nucleotide polymorphism sites （SNPs）in ITS2 region were detected to identify A. calamus and its adulterants. The constructed Neighbour-joining tree can also identify A. calamus, which is consistent with the result of SNPs. The dominant variants in A. calamus possessed approximately 99. 19% in the genome. Only one subordinate variant was detected. The intraspecific and intragenomie divergence of A. calamus are very small. Conclusion： SNPs could be used as an efficiently method to identify A. calamus and its adulterants.

关 键 词：藏菖蒲 DNA条形码 鉴定 ITS2 

分 类 号：R394[医药卫生—医学遗传学]