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      DNA Mini-Barcoding: A Derived Barcoding Method for Herbal Molecular Identification

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          Abstract

          In recent years, the demand for natural herbal products (NHP) has increased; however, the quality of these products is difficult to confirm due to the lack of a comprehensive quality control system. Traditional methods are not effective in detecting processed ingredients. DNA barcoding is an established technique that has been used for more than 10 years. This technique uses short standard sequences (generally 200–600 bp) to identify species. While a complete DNA barcode is difficult to obtain from NHP due to DNA degradation, mini-barcoding is a complementary tool to identify species in NHP. DNA mini-barcoding uses smaller DNA segments for polymerase chain reaction amplification and can be applied to identify species rapidly. The present review summarizes the development and application of DNA mini-barcodes over recent years and discusses the limitations of this technique. This review also compares mini-barcoding and meta-barcoding, a technique using universal polymerase chain reaction primers to simultaneously amplify multiple DNA barcodes and identify many species in a single environmental sample. Additionally, other detection methods that can be combined with mini-barcodes, such as nucleotide signatures, high-resolution DNA melting analysis, and gold nanoparticles, are discussed. DNA mini-barcoding can fill the gaps left by other methods in the field of herbal molecular identification.

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          A DNA-based method for rationally assembling nanoparticles into macroscopic materials.

          Colloidal particles of metals and semiconductors have potentially useful optical, optoelectronic and material properties that derive from their small (nanoscopic) size. These properties might lead to applications including chemical sensors, spectroscopic enhancers, quantum dot and nanostructure fabrication, and microimaging methods. A great deal of control can now be exercised over the chemical composition, size and polydispersity of colloidal particles, and many methods have been developed for assembling them into useful aggregates and materials. Here we describe a method for assembling colloidal gold nanoparticles rationally and reversibly into macroscopic aggregates. The method involves attaching to the surfaces of two batches of 13-nm gold particles non-complementary DNA oligonucleotides capped with thiol groups, which bind to gold. When we add to the solution an oligonucleotide duplex with 'sticky ends' that are complementary to the two grafted sequences, the nanoparticles self-assemble into aggregates. This assembly process can be reversed by thermal denaturation. This strategy should now make it possible to tailor the optical, electronic and structural properties of the colloidal aggregates by using the specificity of DNA interactions to direct the interactions between particles of different size and composition.
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            Gold nanoparticles in chemical and biological sensing.

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              Validation of the ITS2 Region as a Novel DNA Barcode for Identifying Medicinal Plant Species

              Background The plant working group of the Consortium for the Barcode of Life recommended the two-locus combination of rbcL + matK as the plant barcode, yet the combination was shown to successfully discriminate among 907 samples from 550 species at the species level with a probability of 72%. The group admits that the two-locus barcode is far from perfect due to the low identification rate, and the search is not over. Methodology/Principal Findings Here, we compared seven candidate DNA barcodes (psbA-trnH, matK, rbcL, rpoC1, ycf5, ITS2, and ITS) from medicinal plant species. Our ranking criteria included PCR amplification efficiency, differential intra- and inter-specific divergences, and the DNA barcoding gap. Our data suggest that the second internal transcribed spacer (ITS2) of nuclear ribosomal DNA represents the most suitable region for DNA barcoding applications. Furthermore, we tested the discrimination ability of ITS2 in more than 6600 plant samples belonging to 4800 species from 753 distinct genera and found that the rate of successful identification with the ITS2 was 92.7% at the species level. Conclusions The ITS2 region can be potentially used as a standard DNA barcode to identify medicinal plants and their closely related species. We also propose that ITS2 can serve as a novel universal barcode for the identification of a broader range of plant taxa.
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                Author and article information

                Contributors
                Journal
                Front Plant Sci
                Front Plant Sci
                Front. Plant Sci.
                Frontiers in Plant Science
                Frontiers Media S.A.
                1664-462X
                28 August 2019
                2019
                : 10
                : 987
                Affiliations
                Chinese Academy of Medical Sciences & Peking Union Medical College, Institute of Medicinal Plant Development , Beijing, China
                Author notes

                Edited by: Roger Deal, Emory University, United States

                Reviewed by: G. Ravikanth, Ashoka Trust for Research in Ecology and the Environment (ATREE), India; Panagiotis Madesis, Institute of Applied Biosciences (INAB), Greece

                *Correspondence: Jianping Han, jphan@ 123456implad.ac.cn

                This article was submitted to Technical Advances in Plant Science, a section of the journal Frontiers in Plant Science

                Article
                10.3389/fpls.2019.00987
                6724574
                31555305
                d50db6eb-c4c5-4359-883d-fbf8cb3df9e0
                Copyright © 2019 Gao, Liu, Wang, Wei and Han.

                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) and the copyright owner(s) 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.

                History
                : 31 July 2018
                : 12 July 2019
                Page count
                Figures: 1, Tables: 1, Equations: 0, References: 105, Pages: 11, Words: 9221
                Funding
                Funded by: National Natural Science Foundation of China 10.13039/501100001809
                Award ID: 81673552
                Funded by: CAMS Innovation Fund for Medical Sciences
                Award ID: 2016-I2M-3-016
                Categories
                Plant Science
                Review

                Plant science & Botany
                dna mini-barcoding,dna barcoding,molecular identification,natural herbal products,biomarker

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