摘要
目的:评价几条热点DNA条形码候选序列对马鞭草科药用植物的鉴别能力。方法:选择psbA-trnH,rbcL,matK,ITS2和ITS序列进行评价,使用各序列的通用引物进行扩增和测序,用扩增及测序成功率,种内种间差异,barcod-ing gap,基于BLAST 1和Nearest Distance方法的鉴定成功率等指标来评价各序列的鉴定能力。结果:对马鞭草科32个种55个样本的序列分析发现,matK的扩增成功率太低,未纳入后续分析;ITS,ITS2,psbA-trnH以及rbcL的扩增及测序成功率分别为83.6%,83.6%,96.4%,98.2%,其鉴定成功率除rbcL为77.8%,75.9%外都为100%,但ITS2序列的种间,种内差异,barcoding gap较psbA-trnH,ITS有明显优势。ITS2序列进一步纳入网上163个样品的数据后其鉴定成功率依然可以达到89.5%,87.6%。结论:考虑到psbA-trnH较高的测序成功率,ITS2和psbA-trnH是适合马鞭草科植物鉴别的一个较好DNA条形码序列组合。
Objective： To evaluate the ability of several hotspot candidate sequence of DNA barcodes for identifying medicinal plant species in Verbenaceae.Method： Using universal primers,three chloroplast sequences,psbA-trnH,rbcL,matK,two nuclear ribosomal DNA ITS2 and ITS were amplified and sequenced.PCR amplification and sequencing efficiency,intra-and inter-specific variation,barcoding gap and identification efficiency（with BLAST 1 and Nearest Distance methods） were used to evaluate these loci.Result： The rate of successful amplification using matK was too low to further analyze,and the rate of successful amplification and sequencing using psbA-trnH,ITS,and ITS2 and rbcL sequence was 83.6%,83.6%,96.4%,98.2%,respectively.The rate of successful identification using psbA-trnH,ITS,and ITS2 was 100% at the species level except that rbcL was 77.8%,75.9% for 55 samples belonging to 32 species,but ITS2 did better in intra-and inter-specific variation,barcoding gap than the other loci.The rate of successful identification of ITS2 was 89.5%,87.6% even when joining the date of 165 samples from GenBank.Conclusion： It proposes that the combination of ITS2 and psbA-trnH senquence is promising for the identification of the species in Verbenaceae.
出处
《中国中药杂志》
CAS
CSCD
北大核心
2012年第8期1107-1113,共7页
China Journal of Chinese Materia Medica
基金
国际科技合作项目(2007DFA30990)
卫生行业科研专项(200802043)
