Identification of medicinal plant Schisandra chinensis using a potential DNA barcode ITS2

Abstract

IntroductionSchisandra chinensis (Turcz.) Baill. grows mainly in the northeast China and its mature fruits are famous traditional Chinese medicine recorded in "Chinese Pharmacopoeia". As a traditional medicinal herb, S. chinensis has been used as an astringent curing dry cough, asthma, night sweats, nocturnal seminal emissions and chronic diarrhea [1,2]. Modern medical research had proved that S. chinensis contains multiple active components used to protect liver [3][4][5], prevent senility [6][7][8][9], restrain oxidation [10,11], improve human body immunity ability [12], regulate the central nervous system and so on [13,14].Commonly S. chinensis and its adulterants are frequently found in the market together. Schisandra sphenanthera Rehd. et Wils. is important traditional Chinese medicine recorded in "Chinese Pharmacopoeia", too, but its active components of the kinds and levels are clearly different from S. chinensis [15]. Conventional viewpoints are that the medicinal value of S. chinensis is better than S. sphenanthera. But the two medicines are often mistaken for each other [16]. The identification of the two species of Schisandra is difficult when based solely on morphological characteristics. Additionally, some limitations in traditional taxonomy prevent this technique from meeting the complicated demands of species recognition [17]. As such, a method for the simple and accurate authentication of Schisandra is indispensible.As yet some works had been done to differentiation or identification of Schisandra species using RAPD, ISSR, rbcL and ITS [18][19][20]. Recently, being part of ITS, ITS2 was relatively easy to be amplified using one pair of universal primers [21,22]. In addition, ITS2 had been found to provide taxonomic signature in systematic evolution [23,24]. The ITS2 region was also a promising potential molecular marker to be used for rapid taxonomic classification [21,22]. To our best knowledge, applying ITS2 region to identify plant materials from Schisandra with such a large sample size and geographic range had not been reported. And these studies were not found whether the ITS2 region could be used as a genomic marker to identify different Schisandra populations from different ecological environment and geographical distribution. In the current study, we utilized ITS2 as a DNA barcode to distinguish medicinal plants within the Schisandra genus and populations in order to ensure their safe, effective application in traditional using. AbstractTo test whether the internal transcribed spacer 2 (ITS2) region is an effective marker for using in authenticating of the Schisandra chinensis at the species and population levels, separately. And the results showed that the wild populations had higher percentage of individuals that had substitution of C→A at site 86-bp than the cultivated populations. At sites 10-bp, 37-bp, 42-bp and 235-bp, these bases of the Schisandra sphenanthera samples differed from that of S. chinensis. Two species showed higher levels of inter-specific divergence than intra-specif...

Evaluation of Four Commonly Used DNA Barcoding Loci for Chinese Medicinal Plants of the Family Schisandraceae

“…In respect to the authentication of medicinal plants, there were 24 Chinese medicinal plants of Schisandraceae, nine species of Schisandra , three of Kadsura , and 12 of Illicium successfully discriminated via one or more diagnostic methods by single locus or multi-locus combinations in this study ( S9 Table ). Taking important medicinal plants for example, for Schisandra chinensis , the ITS region that could provide more diagnostic characters for this species than other regions was more suitable for its authentication ( S5 – S8 Tables), which has also been supported by the study of Li et al [ 55 ]. For Illicium verum , ITS and matK were more suitable for its authentication, and they could easily distinguish this species from others using the diagnostic characters through visual examination of the alignments ( S5 – S8 Tables).…”

“…The ITS region has been treated as one of the most appropriate DNA barcodes because of its higher variability, which might enhance identification rates even in closely related species [ 12 , 13 ]. Previous studies have suggested that ITS/ITS2 is able to discriminate several Schisandra species [ 55 , 56 ]. In addition, trnH-psbA has been suggested as a promising locus in many studies [ 14 , 95 – 98 ], including some on medicinal plants [ 99 – 101 ].…”

“…Furthermore, ITS2 and ITS distinguish Schisandra chinensis from S . sphenanthera [ 55 ], and S . sphenanthera from its adulterant S .…”

Molecular authentication of Anthemis deserti Boiss. (Asteraceae) based on ITS2 region of nrDNA gene sequence

“…The latest advancement in the DNA sequencing technology and bioinformatics tools for DNA sequence data analysis lead to the development of DNA barcoding techniques (Hebert et al, 2003, Hebert et al, 2004, Hebert and Gregory, 2005, Hebert and Barrett, 2005, Ali et al, 2014) which have revolutionaries the method of the plant taxonomic identification (Poczai and Hyvönen, 2010) using DNA barcoding method especially based on DNA barcode sequence such as nrDNA ITS1, nrDNA ITS2 (Chen et al, 2010, Yao et al, 2010), rbcL, matK, ycf5, rpoC1, psbA-trnH, rps16, trnL-F and ndhF ( Ali et al, 2014). DNA barcoding have successfully employed in authentication of Crocus sativus (Jiang et al, 2014), Schisandra chinensis (Li et al, 2013), Astragalus (Gao et al, 2009). The Next Generation Sequencing (NGS) is comparatively new, and have been demonstrated in the authentication of Costus pictus (Annadurai et al, 2012), Aconitum (Yun et al, 2015), Dendrobium officinale (Guo et al, 2013), Huperzia serrata and Phlegmariurus carinatus (Luo et al, 2010), Valeriana officinalis (Pyle et al, 2012), Hippophae rhamnoides (Ghangal et al, 2013), Ocimum sanctum (Rastogi et al, 2015), Beta vulgaris (Dohm et al, 2014), Panax ginseng (Jayakodi et al, 2014), Elaeis guineensis (Singh et al, 2013), Curcuma longa (Annadurai et al, 2013), Catharanthus roseus (Van Moerkercke et al, 2013), Withania somnifera (Gupta et al, 2013), Azadirachta indica (Krishnan et al, 2012), Cannabis sativa (van Bakel et al, 2011) and Populus trichocarpa (Tuskan et al, 2006).…”

Characteristic and Phylogenetic Analysis of the Complete Chloroplast Genomes of Three Medicinal Plants of Schisandraceae

“…There are currently about 80 species of Schisandraceae, including 34 medicinal plants [ 20 , 21 ]. The commonly used agents in Schisandraceae are found in S. chinensis and S. sphenanthera , and many methods can be used for identification, like chemistry and molecular pharmacy [ 22 – 24 ].…”