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      Discovery of differential sequences for improving breeding and yield of cultivated Ophiocordyceps sinensis through ITS sequencing and phylogenetic analysis

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          Abstract

          To accelerate the breeding process of cultivated Ophiocordyceps sinensis and increase its yield, it is important to identify molecular fingerprint of dominant O. sinensis. In the present study, we collected 3 batches of industrially cultivated O. sinensis product with higher yield than the others and compared their internal transcribed spacer (ITS) sequences with the wild and the reported. The ITS sequence was obtained by bidirectional sequencing and analyzed with molecular systematics as a DNA barcode for rapid and accurate identification of wild and cultivated O. sinensis collected. The ITS sequences of O. sinensis with detailed collection loci on NCBI were downloaded to construct a phylogenetic tree together with the sequences obtained from the present study by using neighbor-joining method based on their evolution relationship. The information on collection loci was analyzed with ArcGIS 10.2 to demonstrate the geographic distribution of these samples and thus to determine the origin of the dominant samples. The results showed that all wild and cultivated samples were identified as O. sinensis and all sequences were divided into seven phylogenetic groups in the tree. Those groups were precisely distributed on the map and the process of their system evolution was clearly presented. The three cultivated samples were clustered into two dominant groups, showing the correlation between the industrially cultivated samples and the dominant wild samples, which can provide references for its optimized breeding in the future.

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          Most cited references 9

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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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            The Scientific Rediscovery of an Ancient Chinese Herbal Medicine:Cordyceps sinensisPart I

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              Caterpillar Fungus (Ophiocordyceps sinensis) Production and Sustainability on the Tibetan Plateau and in the Himalayas

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                Author and article information

                Journal
                CJNM
                Chinese Journal of Natural Medicines
                Elsevier
                1875-5364
                20 October 2018
                : 16
                : 10
                : 749-755
                Affiliations
                1State Key Laboratory of Quality Research in Chinese Medicine (Macau University of Science and Technology), Avenida Wailong, Taipa, Macau Special Administrative Region, China
                2Macau Institute for Applied Research in Medicine and Health, Avenida Wailong, Taipa, Macau Special Administrative Region, China
                3Faculty of Chinese Medicine, Macau University of Science and Technology, Avenida Wailong, Taipa, Macau Special Administrative Region, China
                4Sunshine Lake Pharma Co., Ltd., Dongguan 523808, China
                Author notes
                *Corresponding author: ZHOU Hua, E-mail: hzhou@ 123456must.edu.mo

                ΔThese authors contributed to this work equally.

                These authors have no conflict of interest to declare.

                Article
                S1875-5364(18)30114-6
                10.1016/S1875-5364(18)30114-6
                Copyright © 2018 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.
                Funding
                Funded by: Macao Science and Technology Development Fund
                Award ID: 062/2017/A2
                This work was supported by the Macao Science and Technology Development Fund (No. 062/2017/A2).

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