| Abstract:Objective:Develop specific DNA barcodes based on the chloroplast genome sequence analysis of the original plants of Patrinia medicinal materials to accurately identify Patrinia scabiosaefolia and related species. Methods:The chloroplast genomes of the plants of patrinia, including Patrinia villosa, P.scabiosaefolia and P.heterophylla were sequenced using Illumina high-throughput sequencing technology. Bioinformatics software was employed for genome assembly, annotation, feature analysis, sequence comparison, and phylogenetic analysis. High mutation regions in the chloroplast genome were identified, and specific DNA barcodes were constructed for validation. Results:The chloroplast genomes of three species of Patrinia plants exhibit a tetrad structure, with a total length of 159 585 bp, 158 919 bp, and 158 919 bp, respectively. They encode 130, 132, and 132 genes, including 8 rRNA genes and 37 tRNA genes. The protein coding genes are 85, 87, and 87. ccsA, ndhG, rpl23, and ndhF primers can be used as specific DNA barcodes to successfully amplify 16 samples, accurately clustering Patrinia scabiosaefolia, Patrinia villosa, and Patrinia monandra were clustered in the same branch, while Patrinia heterophylla Bunge and Patrinia scabra Bunge were clustered in a different branch.The ccsA, ndhG, and ndhF sequences can further distinguish Patrinia heterophylla Bunge from Patrinia scabra Bunge. Conclusion:The ccsA, ndhG, rpl23, and ndhF sequences can serve as supplementary specific DNA barcodes to distinguish Patrinia scabiosaefolia and related species, providing a basis for the classification of Patrinia scabiosaefolia and its close relatives. |
