[Show abstract][Hide abstract] ABSTRACT: Rhizosphere and root-associated microbial communities are known to be related to soil-borne disease and plant health. In the present study, the microbial communities in rhizosphere soils and roots of both healthy and diseased Panax notoginseng were analyzed by high-throughput sequencing of 16S rRNA for bacteria and 18S rRNA internal transcribed spacer for fungi, to reveal the relationship of microbial community structure with plant health status. In total, 5593 bacterial operational taxonomic units (OTUs) and 963 fungal OTUs were identified in rhizosphere soils, while 1794 bacterial and 314 fungal OTUs were identified from root samples respectively. Principal coordinate analysis separated the microbial communities both in the rhizosphere soils and roots of diseased P. notoginseng from healthy plants. Compared to those of healthy P. notoginseng, microbial communities in rhizosphere soils and roots of diseased plants showed a decrease in alpha diversity. By contrast, bacterial community dissimilarity increased and fungal community dissimilarity decreased in rhizosphere soils of diseased plants, while both bacterial and fungal community dissimilarity in roots showed no significant difference between healthy and diseased plants. Redundancy analysis at the phylum level showed that mycorrhizal colonization and soil texture significantly affected microbial community composition in rhizosphere soils, whereas shoot nutrition status had a significant effect on microbial community composition in root samples. Our study provided strong evidence for the hypothesis that microbial diversity could potentially serve as an indicator for disease outbreak of medicinal plants, and supported the ecological significance of microbial communities in maintaining plant healthy and soil fertility.
Antonie van Leeuwenhoek 08/2015; DOI:10.1007/s10482-015-0560-x · 2.14 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Tripterygium wilfordii Hook.F. is one of the most valuable medicinal plants because it contains a large variety of active terpenoid compounds, including triptolide, celastrol and wilforlide. All of the pharmacologically active secondary metabolites are synthesized from the 2-C-methyl-D-erythritol 4-phosphateand mevalonate pathway in the isoprenoid biosynthetic system. The key step in this pathway is the isomerization of dimethylallyl diphosphate and isopentenyl diphosphate, which is catalyzed by isopentenyl-diphosphate-isomerase (IPP). In the present study, a full-length cDNA encoding IPI (designate as TwIPI, GenBank accession no.KT279355) was cloned from a suspension of cultured cells from T. wilfordii. The full-length cDNA of TwIPI was 1564 bp and encoded a polypeptide of 288 amino acids. The bioinformatics analysis showed that the deduced TwIPI sequence contained the TNTCCSHPL and WGEHELDY motif. The transcription level of the TwIPI in the suspension cells increased almost five-fold after treatment with methyl jasmonate as an elicitor. A functional color assay in Escherichia coli indicated that TwIPI could promote the accumulation of lycopene and encoded a functional protein. This article is protected by copyright. All rights reserved.
This article is protected by copyright. All rights reserved.
Biotechnology and Applied Biochemistry 08/2015; DOI:10.1002/bab.1427 · 1.32 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Angelica sinensis (Apiaceae) is an endangered alpine herb that is widely used as a medicinal plant in traditional Chinese medicine (TCM). Wild populations of A. sinensis have become quite rare in China. Thus, population genetics studies of this species are urgently needed for its effective conservation and sustainable use. However, to date, no microsatellite loci have been isolated in A. sinensis. To address this issue, we isolated 18 polymorphic loci and genotyped 120 individuals collected from 6 populations. The number of alleles per locus ranged from 1.2 to 5.5, and the average was 2.4. The observed and expected heterozygosity per locus for a population varied, respectively, from 0.000 to 0.983 (averaged at 0.198) and from 0.066 to 0.661 (averaged at 0.333). Deviation from the HardyeWeinberg equilibrium (p < 0.01) was observed for 4 to 14 loci in various populations. These microsatellite markers were cross-amplified in 10 species affinis, and 7 loci were successfully amplified in all species. These microsatellite markers are useful for genetic studies, the conservation management of A. sinensis, and identification of A. sinensis.
[Show abstract][Hide abstract] ABSTRACT: DNA barcoding is a promising species identification method, but it has proved difficult to find a standardized DNA marker in plant. Although the ITS/ITS2 RNA transcript has been proposed as the core barcode for seed plants, it has been criticized for being too conserved in some species to provide enough information or too variable in some species to align it within the different taxa ranks. We selected 30 individuals, representing 16 species and four families, to explore whether ITS2 can successfully resolve species in terms of secondary structure. Secondary structure was predicted using Mfold software and sequence-structure was aligned by MARNA. RNAstat software transformed the secondary structures into 28 symbol code data for maximum parsimony (MP) analysis. The results showed that the ITS2 structures in our samples had a common four-helix folding type with some shared motifs. This conserved structure facilitated the alignment of ambiguous sequences from divergent families. The structure alignment yielded a MP tree, in which most topological relationships were congruent with the tree constructed using nucleotide sequence data. When the data was combined, we obtained a well-resolved and highly supported phylogeny, in which individuals of a same species were clustered together into a monophyletic group. As a result, the different species that are often referred to as the herb "Mu tong" were successfully identified using short fragments of 250 bp ITS2 sequences, together with their secondary structure. Thus our analysis strengthens the potential of ITS2 as a promising DNA barcode because it incorporates valuable secondary structure information that will help improve discrimination between species.
PLoS ONE 07/2015; 10(7):e0131185. DOI:10.1371/journal.pone.0131185 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Tetrabutylphosphonium hydroxide (TBPH) aqueous solution, a novel ionic liquid that could dissolve cellulose rapidly at ambient temperature (25 °C), was used for the first time to develop an extraction method for salidroside from Rhodiola crenulata, used as the model sample, with infrared-assisted extraction (IRAE) in this paper. IRAE-TBPH procedures were optimized using a series of single-factor experiments and under the optimal conditions, the IRAE-TBPH technique not only took a shorter time (from 1.0 h to 8 min) but also afforded a higher extraction rate of salidroside from the herbs (increased by 15.41-38.65%) compared with other extraction techniques, such as TBPH-based heat reflux extraction (HRE-TBPH), ultrasound-assisted extraction (UAE-TBPH) and conventional solvent (methanol, ethanol and pure water) based IRAE. The results indicated IRAE-TBPH to be a fast and efficient extraction technique. Furthermore, the mechanism of IRAE-TBPH was preliminarily studied by means of surface structures and chemical compositions of samples before and after different extraction techniques. On the basis of the destruction of herb surface microstructures, cellulose dissolving property of TBPH and high efficiency heating of infrared irradiation in IRAE-TBPH process, the IRAE-TBPH technique eventually got the maximum yield value. Therefore, TBPH solution as a novel, effective and alternative solvent with higher extraction efficiency in the IRAE of active compounds from medicinal plants showed a great promising prospect.
[Show abstract][Hide abstract] ABSTRACT: Color variation in sea cucumber is one of the most crucial traits affecting price and taste in East Asian countries. However, the relationship and taxonomic status of the three color variants are still unclear. We used 14 samples that covered all three color variants and their geographic distributions, to construct the first phylogeny for the color variants based on the complete mitochondrial genome sequence and a number of tree-building methods (maximum parsimony (MP), maximum likelihood (ML), and Bayesian inference (BI)). The divergence times within color variants were estimated by the Bayesian molecular clock approach using the BEAST program. Our results showed that the color variants were not monophyletic in the well-resolved phylogenetic tree, which strongly refuted their separate species status. The molecular dating estimate revealed that the sea cucumber was a young group, which originated in the early Miocene period (22.03 mya) and rapidly diverged after the late Miocene period. It is interesting that individuals within each variant or geographic distribution were not always closely related and thus did not share a common origin. We propose that although they differ in body color, the three color morphs all belong to a single species of Apostichopus japonicus and the historical marine climate and the hydrographic complexity of the ocean currents could be responsible for their present distribution patterns.
Mitochondrial DNA 05/2015; DOI:10.3109/19401736.2015.1022765 · 1.70 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Ginseng, which is the root of Panax ginseng (Araliaceae), has been used in Oriental medicine as a stimulant and dietary supplement for more than 7,000 years. Older ginseng plants are substantially more medically potent, but ginseng age can be simulated using unscrupulous cultivation practices. Telomeres progressively shorten with each cell division until they reach a critical length, at which point cells enter replicative senescence. However, in some cells, telomerase maintains telomere length. In this study, to determine whether telomere length reflects ginseng age and which tissue is best for such an analysis, we examined telomerase activity in the main roots, leaves, stems, secondary roots and seeds of ginseng plants of known age. Telomere length in the main root (approximately 1 cm below the rhizome) was found to be the best indicator of age. Telomeric terminal restriction fragment (TRF) lengths, which are indicators of telomere length, were determined for the main roots of plants of different ages through Southern hybridization analysis. Telomere length was shown to be positively correlated with plant age, and a simple mathematical model was formulated to describe the relationship between telomere length and age for P. ginseng.
[Show abstract][Hide abstract] ABSTRACT: DNA barcoding has been proposed to be one of the most promising tools for accurate and rapid identification of taxa. However, few publications have evaluated the efficiency of DNA barcoding for the large genera of flowering plants. Dendrobium, one of the largest genera of flowering plants, contains many species that are important in horticulture, medicine and biodiversity conservation. Besides, Dendrobium is a notoriously difficult group to identify. DNA barcoding was expected to be a supplementary means for species identification, conservation and future studies in Dendrobium. We assessed the power of 11 candidate barcodes on the basis of 1,698 accessions of 184 Dendrobium species obtained primarily from mainland Asia. Our results indicated that five single barcodes, i.e., ITS, ITS2, matK, rbcL and trnH-psbA, can be easily amplified and sequenced with the currently established primers. Four barcodes, ITS, ITS2, ITS+matK, and ITS2+matK, have distinct barcoding gaps. ITS+matK was the optimal barcode based on all evaluation methods. Furthermore, the efficiency of ITS+matK was verified in four other large genera including Ficus, Lysimachia, Paphiopedilum, and Pedicularis in this study. Therefore, we tentatively recommend the combination of ITS+matK as a core DNA barcode for large flowering plant genera.
PLoS ONE 01/2015; 10(1):e0115168. DOI:10.1371/journal.pone.0115168 · 3.23 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Deer antler is a precious animal-sourced traditional Chinese medicine. We aimed to rapidly assess the quality of deer antler slices by electronic nose so that we can ensure medical safety. In this study, response intensity of the electronic nose was favorably optimized, and samples were well assessed by using an electronic nose based on LDA model. The results obtained herein suggested that electronic nose could be an effective method to rapidly as- sess the quality of deer antler slices, and could also be an important tool for categorization of complex aroma mixtures for the control of quality of drugs or food.
Revista Brasileira de Farmacognosia 12/2014; 22(6). DOI:10.1016/j.bjp.2014.10.011 · 0.80 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Abstract Antelope horn is a valuable Chinese traditional medicine and widely used in clinic. However, with the deterioration of antelope's living environment and a lot of killing, the saiga population begins falling and in some places plummet. Since the increasing demand of this expensive and good bioactive medicine, the horn of artiodactyla animals is often used as the antelope horn. The adulterated or impostor not only cause damage to clinical medicine but also affect the antelope resources protection and sustainable development. Here, in order to establish a melting curve analysis (MCA) method to distinguish the antelope horn from other animal horns and identify the decoction pieces and Chinese patent medicine in a fast and easy way, animal horns and its decoction pieces, Chinese patent medicines were collected from the market and the DNA of all the collected samples were extracted. The melting curve of two universal fragments (COI and Cyt b) was scanned and Cyt b was selected as feasibility fragment for identifying authentic antelope horn from eight adulterant animal horns. After optimizing the condition for MCA, inspecting the precision and the replication of the method, a reference melting curve modern was established and we performed MCA on the antelope horns, fakes, and adulterants on a 1:1 mix, decoction pieces, and Chinese patent medicine. Thus, this study provides fast and easy methods so that MCA can detect the truth, fakes, and adulterations of antelope horns.
Mitochondrial DNA 12/2014; DOI:10.3109/19401736.2014.989500 · 1.70 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: R2R3-MYB proteins are involved in the primary and secondary metabolism, developmental processes and the responses to biotic and abiotic stresses. Little is known about the functions of R2R3-MYB proteins in Scutellaria baicalensis Georgi which is a traditional Chinese medicinal plants. In this study, the function of a S. baicalensis R2R3-MYB protein, SbMYB8, was investigated. SbMYB8 had similar expression pattern with SbC4H and SbCHS in ABA-treated S. baicalensis, indicating that SbMYB8 might be involved in the flavonoid metabolism. SbMYB8 protein could bind to the GmMYB92 BS3 sequence of SbCHS promoter region, regulating the expression of SbCHS. The SbMYB8 protein was localized to the nucleus where it activated transcription. The transgenic tobacco plants over-expressing SbMYB8 had higher caffeoylquinic acid contents, compared to that in wild type plants. Overexpression of SbMYB8 also changed the expression level of some flavonoid biosynthesis-related genes. It was found that overexpression of SbMYB8 can improve stress tolerance of transgenic plants, and can alter the activity and expression levels of some antioxidant enzymes. These results indicate that SbMYB8 plays important roles in flavonoid biosynthesis and stress tolerance of plant.
Plant Cell Tissue and Organ Culture 12/2014; 120(3). DOI:10.1007/s11240-014-0686-y · 2.61 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Dummy molecularly imprinted polymers (DMIPs) for simultaneously selective removal and enrichment of ginkgolic acids (GAs) during the processing of Ginkgo biloba leaves have been prepared. Two dummy template molecule with similar structural skeleton to GAs, 6-methoxysalicylic acid (MOSA, DT-1) and 6-hexadecyloxysalicylic acid (HOSA, DT-2), have been designed and synthesized. The performance of the DMIPs and NIPs were evaluated including selective recognition capacity, adsorption isotherm, and adsorption kinetics. The selective recognition capacity of the three GAs with four analogues on the sorbents illustrated that the DMIPs sorbents have high specificity for GAs. An efficient method based on DMIP-HOSA coupled with solid-phase extraction (SPE) was developed for simultaneously selective removal and enrichment of ginkgolic acids (GAs) during the processing of Ginkgo biloba leaves. The method showed excellent recoveries (82.5-88.7%) and precision (RSD 0.5-2.6%, n=5) for licorice extracts, Gastrodia elata extracts and pepper extracts spiked at three concentration levels each (50, 100, 200 mu g mL(-1)). The results indicated that GAs and standardized Ginkgo biloba leaves extracts could be obtained simultaneously through the DMIP-SPE.
Journal of Chromatography A 11/2014; 1368. DOI:10.1016/j.chroma.2014.09.070 · 4.26 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Evaluating the safety of traditional medicinal herbs and their major active constituents is critical for their widespread usage. Geniposide, a major active constituent with a defined structure from the traditional medicinal herb Gardenia jasminoides ELLIS fruit, exhibits remarkable anti-inflammatory, antiapoptotic, and antifibrotic properties and has been used in a variety of medical fields, mainly for the treatment of liver diseases. However, geniposide-induced hepatotoxicity and methods for the early detection of hepatotoxicity have yet to be reported. In this study, geniposide-induced hepatotoxicity was investigated. In addition, candidate biomarkers for the earlier detection of geniposide-induced hepatotoxicity were identified using a label-free quantitative proteomics approach on a geniposide overdose-induced liver injury in a rat model. Using an accurate intensity-based, absolute quantification (iBAQ)-based, one-step discovery and verification approach, a candidate biomarker panel was easily obtained from individual samples in response to different conditions. To determine the biomarkers' early detection abilities, five candidate biomarkers were selected and tested using enzyme-linked immunosorbent assays (ELISAs). Two biomarkers, glycine N-methyltransferase (GNMT) and glycogen phosphorylase (PYGL), were found to indicate hepatic injuries significantly earlier than the current gold standard liver biomarker. This study provides a first insight into geniposide-induced hepatotoxicity in a rat model and describes a method for the earlier detection of this hepatotoxicity, facilitating the efficient monitoring of drug-induced hepatotoxicity.
Journal of Proteome Research 10/2014; 13(12). DOI:10.1021/pr5007119 · 5.00 Impact Factor
[Show abstract][Hide abstract] ABSTRACT: Scutellaria baicalensis Georgi is an herbaceous perennial plant used as one of the staple Chinese herbal medicines in China with a long officinal history. However, research on S. baicalensis is currently limited due to the lack of genome and gene expression information. A full-length cDNA library from leaves and roots of S. baicalensis subjected to water deficit and heat, conditions that have been shown to affect baicalein accumulation, was constructed. There were 6491 expressed sequence tags (ESTs) obtained. UniGenes were assembled by BLAST similarity searches and annotated with Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG). A total of 78 simple sequence repeats (SSRs) were identified and SSR markers associated with the active ingredients of S. baicalensis were selected. EST-SSR transferability was determined from 5 populations from different areas. This study is the first to produce a large volume of gene expression data from S. baicalensis to facilitate gene discovery in S. baicalensis and provide an important resource for molecular genetic and functional genomic studies in this species.
[Show abstract][Hide abstract] ABSTRACT: Saffron (Crocus sativus L.) is one of the most important and expensive medicinal spice products in the world. Because of its high market value and premium price, saffron is often adulterated through the incorporation of other materials, such as Carthamus tinctorius L. and Calendula officinalis L. flowers, Hemerocallis L. petals, Daucus carota L. fleshy root, Curcuma longa L. rhizomes, Zea may L., and Nelumbo nucifera Gaertn. stigmas. To develop a straightforward, nonsequencing method for rapid, sensitive, and discriminating detection of these adulterants in traded saffron, we report here the application of a barcoding melting curve analysis method (Bar-MCA) that uses the universal chloroplast plant DNA barcoding region trnH-psbA to identify adulterants. When amplified at DNA concentrations and annealing temperatures optimized for the curve analysis, peaks were formed at specific locations for saffron (81.92°C) and the adulterants: D. carota (81.60°C), C. tinctorius (80.10°C), C. officinalis (79.92°C), Dendranthema morifolium (Ramat.) Tzvel. (79.62°C), N. nucifera (80.58°C), Hemerocallis fulva (L.) L. (84.78°C), and Z. mays (84.33°C). The constructed melting curves for saffron and its adulterants have significantly different peak locations or shapes. In conclusion, Bar-MCA could be a faster and more cost-effective method to authenticate saffron and detect its adulterants.
BioMed Research International 10/2014; 2014:809037. DOI:10.1155/2014/809037 · 2.71 Impact Factor