Genetic Authentication of the Medicinal Plant Portulaca oleracea Using a Quick, Precise, and Sensitive Isothermal DNA Amplification Assay

International Journal of Molecular Sciences · 7 authors, 4 centres

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Using the ITS2 region as a molecular marker, researchers designed and optimized a specific LAMP primer set (PO02) that could discriminate PO from closely related Portulaca species and the common adulterant Bacopa monnieri. The LAMP assay showed 100-fold improved sensitivity over standard PCR, with a detection limit of 100 fg, which was enhanced by the addition of a loop primer and DMSO. The method successfully verified the identity of heat-processed PO samples and commercial herbal products, showing complete agreement with microscopic examination. The authors note that LAMP is suitable for homogeneous samples and requires further development for multiplex identification.

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preclinical research

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ORIGINAL PAPER

PMC10341654

Genetic Authentication of the Medicinal Plant Portulaca oleracea Using a Quick, Precise, and Sensitive Isothermal DNA Amplification Assay

International Journal of Molecular Sciences

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ABSTRACT

Abstract Portulaca oleracea (PO) is a commonly known medicinal crop that is an important ingredient for traditional Chinese medicine (TCM) due to its use as a vegetable in the diet. PO has been recorded to be frequently adulterated by other related species in the market of herbal plants, distorting the PO plant identity. Thus, identification of the botanical origin of PO is a crucial step before pharmaceutical or functional food application. In this research, a quick assay named “loop-mediated isothermal amplification (LAMP)” was built for the specific and sensitive authentication of PO DNA. On the basis of the divergences in the internal transcribed spacer 2 (ITS2) sequence between PO and its adulterant species, the LAMP primers were designed and verified their specificity, sensitivity, and application for the PO DNA authentication. The detection limit of the LAMP assay for PO DNA identification specifically was 100 fg under isothermal conditions at 63 °C for 30 min. In addition, different heat-processed PO samples can be applied for use in PO authentication in the LAMP assay. These samples of PO were more susceptible to the effect of steaming in authentication by PCR than boiling and drying treatment. Furthermore, commercial PO samples pursued from herbal markets were used to display their applicability of the developed LAMP analysis for PO postharvest authentication, and the investigation found that approximately 68.4% of PO specimens in the marketplace of herbal remedies were adulterated. In summary, the specific, sensitive, and rapid LAMP assay for PO authentication was first successfully developed herein, and its practical application for the inspection of adulteration in PO samples from the herbal market was shown. This LAMP assay created in this study will be useful to authenticate the botanical origin of PO and its commercial products. Keywords: Portulaca oleracea , medicinal crop, loop-mediated isothermal amplification (LAMP), traditional Chinese medicine (TCM), authentication

CITATION EXPORT

Mo-Rong Xu, Fang-Chun Sun, Bo-Cheng Yang et al.. Genetic Authentication of the Medicinal Plant Portulaca oleracea Using a Quick, Precise, and Sensitive Isothermal DNA Amplification Assay. International Journal of Molecular Sciences. (2023). https://doi.org/10.3390/ijms241310730

AUTHORS

7 · 4 CENTRES

Mo-Rong Xu, Fang-Chun Sun, Bo-Cheng Yang, Hsi-Jien Chen, Chia-Hsin Lin, Jai-Hong Cheng, Meng-Shiou Lee

1Department of Chinese Pharmaceutical Science and Chinese Medicine Resources, China Medical University, Taichung 40402, Taiwan; [email protected] (M.-R.X.); [email protected] (B.-C.Y.); [email protected] (C.-H.L.)
2Department of Medicinal Botanicals and Foods on Health Applications, Da-Yeh University, Changhua 515006, Taiwan; [email protected]
3Department of Safety, Health and Environmental Engineering, Ming Chi University of Technology, New Taipei City 24301, Taiwan; [email protected]
4Center for Shockwave Medicine and Tissue Engineering, Kaohsiung Chang Gung Memorial Hospital, Chang Gung University College of Medicine, Kaohsiung 833, Taiwan; [email protected]