|
| Development and identification of mini-barcode based on chloroplast genome of Isatis indigotica and its adulterants |
| Hits 469 Download times 1751 Received:February 01, 2023 |
| View Full Text View/Add Comment Download reader |
| DOI
10.11656/j.issn.1672-1519.2023.05.21 |
| Key Words
Isatis indigotica;chloroplast genome;primer design;mini-barcode |
| Author Name | Affiliation | E-mail | | GUO Hua | Tianjin University of Traditional Chinese Medicine, State Key Laboratory of Component-Based Chinese Medicine, Tianjin 301617, China
Haihe Laboratory of Modern Chinese Medicine, Tianjin 301617, China | | | XU Liuwei | Tianjin University of Traditional Chinese Medicine, State Key Laboratory of Component-Based Chinese Medicine, Tianjin 301617, China
Haihe Laboratory of Modern Chinese Medicine, Tianjin 301617, China | | | XING Zhimei | Tianjin University of Traditional Chinese Medicine, State Key Laboratory of Component-Based Chinese Medicine, Tianjin 301617, China
Haihe Laboratory of Modern Chinese Medicine, Tianjin 301617, China | | | YANG Xia | Tianjin University of Traditional Chinese Medicine, State Key Laboratory of Component-Based Chinese Medicine, Tianjin 301617, China
Haihe Laboratory of Modern Chinese Medicine, Tianjin 301617, China | | | TIAN Xiaoxuan | Tianjin University of Traditional Chinese Medicine, State Key Laboratory of Component-Based Chinese Medicine, Tianjin 301617, China
Haihe Laboratory of Modern Chinese Medicine, Tianjin 301617, China | tian_xiaoxuan@tjutcm.edu.cn |
|
| Abstract
|
| [Objective] Based on the of Isatis indigotica and its adulterants, highly variable regions were screened and DNA mini-barcode were developed. The identification ability was verified using the original herbs of Isatis indigotica.[Methods] All published chloroplast genome sequences of Isatis indigotica and its adulterants download from GenBank database were imported into PhyloSuite v1.2.1 software to extract their consensus protein-coding genes, subsequent DnaSP 6 software was used to perform nucleotide polymorphism analysis to screen for candidate DNA mini-barcode regions. Through multi-sequence alignment by Geneious, hypervariable regions were used for the development of mini-barcode and primers design. Polymerase chain reaction(PCR) and sequencing were performed on Isatis indigotica to evaluate the identification ability of the mini-barcode by kit method.[Results] Based on the results of the nucleotide polymorphism analysis of chloroplast genome, a specific DNA mini-barcode was developed in the accD region. By genetic distance calculation and the sequencing data analysis, the results showed that the mini-barcode was successfully amplified and identified.[Conclusion] In this study, mini-barcode was designed to identify Isatis indigotica and its adulterants. The results of genetic distance and sequencing data analysis were used to derive the successful primer that can distinguish Isatis indigotica and its adulterants. This study lays a foundation for the identification of the samples which may have degraded or mixed DNA, such as Chinese patent medicines containing Isatis indigotica. |
|
|
|
|
|
|
|
