摘要
To ensure the safety of medications,it is vital to accurately authenticate species of the Apocynaceae family,which is rich in poisonous medicinal plants.We identified Apocynaceae species by using nuclear internal transcribed spacer 2(ITS2)and psb Atrn H based on experimental data.The identification ability of ITS2 and psb A-trn H was assessed using specific genetic divergence,BLAST1,and neighbor-joining trees.For DNA barcoding,ITS2 and psb A-trn H regions of 122 plant samples of 31 species from 19 genera in the Apocynaceae family were amplified.The PCR amplification for ITS2 and psb A-trn H sequences was 100%.The sequencing success rates for ITS2 and psb A-trn H sequences were 81%and 61%,respectively.Additional data involved 53 sequences of the ITS2 region and 38 sequences of the psb A-trn H region were downloaded from Gen Bank.Moreover,the analysis showed that the interspecific divergence of Apocynaceae species was greater than its intra-specific variations.The results indicated that,using the BLAST1 method,ITS2 showed a high identification efficiency of 97%and 100%of the samples at the species and genus levels,respectively,via BLAST1,and psb A-trn H successfully identified 95%and 100%of the samples at the species and genus levels,respectively.The barcode combination of ITS2/psb A-trn H successfully identified 98%and 100%of samples at the species and genus levels,respectively.Subsequently,the neighbor joining tree method also showed that barcode ITS2 and psb A-trn H could distinguish among the species within the Apocynaceae family.ITS2 is a core barcode and psb A-trn H is a supplementary barcode for identifying species in the Apocynaceae family.These results will help to improve DNA barcoding reference databases for herbal drugs and other herbal raw materials.
基金
supported by the CAMS Initiative for Innovative Medicine(No.2016-12M-2-003)
the Special Subsidies for Public Health Services of TCM("The National Survey of TCM Resources",DSS,MOF,No.66/2017)。
