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ORIGINAL RESEARCH article
Front. Plant Sci.
Sec. Functional and Applied Plant Genomics
Volume 15 - 2024 |
doi: 10.3389/fpls.2024.1492083
This article is part of the Research Topic Plant Genotyping: From Traditional Markers to Modern Technologies, Volume II View all articles
Single nucleotide polymorphism (SNP)-based visual identification of Rhodiola crenulata using loop-mediated isothermal amplification (LAMP) technique
Provisionally accepted
Li Hao
1*
Xin Shi
1
Shiyu Wen
1*
Caiye Yang
1*
Yaqi Chen
1*
Samo Yue
1*
Jiaqiang Chen
1*
Kexin Luo
1*
Bingliang Liu
1
Yanxia Sun
1*
Yi Zhang
2*- 1 Chengdu University, Chengdu, Sichuan Province, China
- 2 Chengdu Institute of Biology, Chinese Academy of Sciences (CAS), Chengdu, China
Rhodiola crenulata (Hook.f. & Thomson) H.Ohba, a member of the Crassulaceae family, is a traditional Chinese medicine recognized as the original source of Rhodiolae Crenulatae Radix et Rhizoma in the 2020 edition of the China Pharmacopoeia. It has been widely used in both Asia and Europe to enhance stress resistance and reduce fatigue. However, the classification of Rhodiola species can lead to confusion, raising safety concerns in the herbal medicine market. In this study, we aimed to differentiate Rh. crenulata from two closely related species. We initially identified single nucleotide polymorphism (SNP) sites using cleaved amplified polymorphic sequence (CAPS) RT-PCR. Based on the identified SNP sites, we designed specific loop backward primers and evaluated optimal reaction temperatures using real-time fluorescent loop-mediated isothermal amplification (LAMP) to identify Rh. crenulata. The final detection system exhibited a sensitivity of up to 1,000 copies of the target DNA, maintaining a constant reaction temperature of 62℃ within 35 minutes. To facilitate visualization, we incorporated two color indicators, hydroxy naphthol blue (HNB) and neutral red (N-red), into the reaction system. Collectively, we developed a simple, rapid, specific, sensitive, and visible method to distinguish Rh. crenulata from other two Rhodiola species and Rh. crenulata hybrids. This approach has significant potential for applications in pharmaceutical industry.
Keywords: Medicinal herb, original plant identification, molecular authentication method, 21 naked-eye detection, ITS sequences, using only visual inspection enhance the reaction rate
Received: 16 Sep 2024; Accepted: 23 Dec 2024.
Copyright: © 2024 Hao, Shi, Wen, Yang, Chen, Yue, Chen, Luo, Liu, Sun and Zhang. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence:
Li Hao, Chengdu University, Chengdu, 610106, Sichuan Province, China
Shiyu Wen, Chengdu University, Chengdu, 610106, Sichuan Province, China
Caiye Yang, Chengdu University, Chengdu, 610106, Sichuan Province, China
Yaqi Chen, Chengdu University, Chengdu, 610106, Sichuan Province, China
Samo Yue, Chengdu University, Chengdu, 610106, Sichuan Province, China
Jiaqiang Chen, Chengdu University, Chengdu, 610106, Sichuan Province, China
Kexin Luo, Chengdu University, Chengdu, 610106, Sichuan Province, China
Yanxia Sun, Chengdu University, Chengdu, 610106, Sichuan Province, China
Yi Zhang, Chengdu Institute of Biology, Chinese Academy of Sciences (CAS), Chengdu, China
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