Chemical and Genetic Discrimination of Cistanches Herba Based on UPLC-QTOF/MS and DNA Barcoding

Zheng, Shu-Jian; Jiang, Xue; Wu, Labin; Wang, Zenghui; Huang, Linfang

doi:10.1371/journal.pone.0098061

Cited by 41 publications

(28 citation statements)

References 20 publications

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“…Two plastid barcodes (the coding genes matK and rbcL ) and the nuclear ribosomal internal transcribed spacer ( ITS ) were amplified according to PCR reaction conditions following guidelines from the Consortium of Barcode of Life (CBOL) plant-working group, and sequenced using universal primers. The psbA - trnH intergenic spacer region was tested with two different primer sets: fwdPA and revTH [ 39 ] and psbA and trnH [ 23 ] corresponding to species range, but unsuccessful amplification led to its omission from the present study ( Table 1 ). The selected DNA regions were amplified by using a standard PCR, whereby amplification for each primer set was carried out in a 50-μl volume solution containing 1X Taq DNA polymerase buffer, 200 μM each dNTP (dATP:dTTP:dCTP:dGTP in 1:1:1:1 parts), 10 pmol of each primer (forward and reverse), 1 unit of Taq DNA polymerase and 25 ng of template DNA.…”

Section: Methodsmentioning

confidence: 99%

Evaluation of single and multilocus DNA barcodes towards species delineation in complex tree genus Terminalia

et al. 2017

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DNA barcoding is used as a universal tool for delimiting species boundaries in taxonomically challenging groups, with different plastid and nuclear regions (rbcL, matK, ITS and psbA-trnH) being recommended as primary DNA barcodes for plants. We evaluated the feasibility of using these regions in the species-rich genus Terminalia, which exhibits various overlapping morphotypes with pantropical distribution, owing to its complex taxonomy. Terminalia bellerica and T. chebula are ingredients of the famous Ayurvedic Rasayana formulation Triphala, used for detoxification and rejuvenation. High demand for extracted phytochemicals as well as the high trade value of several species renders mandatory the need for the correct identification of traded plant material. Three different analytical methods with single and multilocus barcoding regions were tested to develop a DNA barcode reference library from 222 individuals representing 41 Terminalia species. All the single barcodes tested had a lower discriminatory power than the multilocus regions, and the combination of matK+ITS had the highest resolution rate (94.44%). The average intra-specific variations (0.0188±0.0019) were less than the distance to the nearest neighbour (0.106±0.009) with matK and ITS. Distance-based Neighbour Joining analysis outperformed the character-based Maximum Parsimony method in the identification of traded species such as T. arjuna, T. chebula and T. tomentosa, which are prone to adulteration. rbcL was shown to be a highly conservative region with only 3.45% variability between all of the sequences. The recommended barcode combination, rbcL+matK, failed to perform in the genus Terminalia. Considering the complexity of resolution observed with single regions, the present study proposes the combination of matK+ITS as the most successful barcode in Terminalia.

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Section: Methodsmentioning

confidence: 99%

Evaluation of single and multilocus DNA barcodes towards species delineation in complex tree genus Terminalia

et al. 2017

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“…Noncoding psbA‐trnH region of cpDNA was amplified using universal primers fwd‐PA: 5′‐GTT ATG CAT GAA CGT AAT GCT C‐3′ and rev‐TH: 5′‐CGC GCA TGG TGG ATT CAC AAT CC‐3′ (Newmaster & Ragupathy, ; Zheng, Jiang, Wu, Wang, & Huang, ). PCR was run in a manner similar to that of ISSR‐PCR with the following cycling parameters: an initial denaturation at 94°C for 5 min; 30 cycles of denaturation at 94°C for 1 min, annealing at 55°C for 1 min, extension at 72°C for 90 s; final extension at 72°C for 7 min.…”

Section: Methodsmentioning

confidence: 99%

Higher efficiency of ISSR markers over plastid psbA‐trnH region in resolving taxonomical status of genus Ocimum L.

Kumar

Mishra

Baskaran

et al. 2016

Ecology and Evolution

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High level of morphological as well as chemical variability exists within the genus Ocimum, and its taxonomy and phylogenetic relationships are still doubtful. For evaluating interspecific genetic relationships among the Ocimum species, genotyping with intersimple sequence repeat (ISSR) markers and sequence analyses of noncoding psbA‐trnH intergenic region belonging to chloroplast DNA were carried out. Although ISSR markers are highly efficient and reproducible, they have not been used extensively in phylogenetic studies. The use of the plastidial barcode candidate was expected to provide more variable and informative insight into evolutionary rates, and was thus employed as a phylogenetic marker to assess interspecific relationships. This study revealed that the ISSR markers were more efficient than psbA‐trnH sequences in resolving the current status of Ocimum L. genus. Distance‐ and character‐based methodological approaches applied on the molecular data with biparental and maternal inheritance were used for deducing the phylogenetic relationships among Ocimum species. Average polymorphic information content (0.344) and resolving power (6.285) depicted through ISSR markers proved to be efficient in discriminating the studied species of Ocimum. The primers used in this study revealed 99.585% polymorphism across the species demonstrating the polymorphic nature of ISSR markers.

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“…The dried roots of B. malconense were collected from Sichuan Province, China. The plant material was identified and authenticated by Dr. Linfang Huang based on sequences of the plastid psbA-trnH intergenic region [ 13 , 14 ]. The sequences of plant material (ZSH1 and ZSH2) highly matched the sequence of B. malconense (GenBank accession: JN788921) (Additional file 2 ).…”

Section: Methodsmentioning

confidence: 99%

A novel coumarin, (+)-3′-angeloxyloxy-4′-keto-3′,4′-dihydroseselin, isolated from Bupleurum malconense (Chaihu) inhibited NF-κB activity

Lin

Huang

et al. 2016

Chin Med

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BackgroundThis study aims to identify the major anti-inflammatory components in the petroleum ether extract of Bupleurummalconense (Chaihu), by bioassay-guided fractionation, and to investigate the anti-inflammatory mechanisms of active components in lipopolysaccharide (LPS)-stimulated murine macrophage RAW-Blue cells.MethodsA QUANTI-Blue assay was used to guide fractionation of B.malconense root extract. The petroleum ether extract which exerted significant secreted embryonic alkaline phosphatase (SEAP) inhibition effect was purified by silica gel column chromatography and assisted with reverse phase HPLC. The major bioactive compound which significantly inhibited SEAP activity was obtained and its anti-inflammatory effects in LPS-induced RAW-Blue cells were measured by the overproduction of NO (Griess method), gene expression of Il-1β, Tnf-α and iNos (real-time PCR). In parallel, protein expressions of COX-2, iNOS and IκB-α were determined by western blot.ResultsIn bioassay-guided fractionation using LPS-stimulated mouse macrophage RAW-Blue cells, (+)-3′-angeloxyloxy-4′-keto-3′,4′-dihydroseselin (Pd-Ib) was identified by MS and NMR spectral analyses. Pd-Ib (5, 10, 20 μg/mL) suppressed the gene expression of Il-1β (P < 0.0001, P < 0.0001, P < 0.0001 for three respective concentrations), Tnf-α (P = 0.006, P = 0.001, P < 0.0001 for three respective concentrations) and iNos (P = 0.009, P < 0.0001, P < 0.0001 for three respective concentrations) in LPS-stimulated macrophages. The production of cyclooxygenase-2 (P = 0.019, P = 0.002, P < 0.0001), iNOS (P < 0.0001, P < 0.0001, P < 0.0001 for three respective concentrations) and NO (P < 0.0001, P < 0.0001, P < 0.0001 for three respective concentrations) significantly decreased when macrophages were treated with Pd-Ib (5, 10, 20 μg/mL) in the presence of LPS. Pd-Ib (5, 10, 20 μg/mL) suppressed the nuclear activation of NF-κB while it up-regulated the IκB-α level (P = 0.028, P = 0.013, P = 0.005 for three respective concentrations) in LPS-stimulated macrophages.ConclusionsPd-Ib isolated from B.malconense suppressed LPS-induced inflammatory responses in macrophages by inhibiting NF-κB activity and reducing the expression of iNOS, COX-2 as well as pro-inflammatory cytokines.Electronic supplementary materialThe online version of this article (doi:10.1186/s13020-016-0077-x) contains supplementary material, which is available to authorized users.

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Chemical and Genetic Discrimination of Cistanches Herba Based on UPLC-QTOF/MS and DNA Barcoding

Cited by 41 publications

References 20 publications

Evaluation of single and multilocus DNA barcodes towards species delineation in complex tree genus Terminalia

Evaluation of single and multilocus DNA barcodes towards species delineation in complex tree genus Terminalia

Higher efficiency of ISSR markers over plastid psbA‐trnH region in resolving taxonomical status of genus Ocimum L.

A novel coumarin, (+)-3′-angeloxyloxy-4′-keto-3′,4′-dihydroseselin, isolated from Bupleurum malconense (Chaihu) inhibited NF-κB activity

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