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Liang J.,Huaihua University | Liang J.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Li S.-H.,Huaihua University | Li S.-H.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Chen C.-Q.,Huaihua University
Chinese Traditional and Herbal Drugs | Year: 2014

Objective: To study the chemical constituents from the whole plant of Swertia davidi. Methods: The chemical constituents were isolated and purified with silica gel column chromatography, silica gel chromatography, etc. The structures of the compounds were identified by physicochemical properties and various spectroscopic methods. Results: Sixteen compounds were isolated as uvaol (1), oleanolic acid (2), 5-carboxyl-3, 4-dihydrogen-1H - 2-benzopyran-1-one (3), 3β, 12β-pregnane-16-12-ene-20 (4), swertisin (5), 6-C-β-glucopyranosyl-7-O-methylluteolinidin (6), isoscoparin (7), 6-demethoxy-7-methylcapillarisin (8), 3-acetoxy-28-hydro-xyl-12-eneurs (9), gentiopicroside (10), N-pentacosy-2-carboxy-benzoylamide (11), 4-O-glycosyloxy-2-hydroxy-6-methoxy-acetophenone (12), quercetin-3-O-β-D-xylopyranosyl-(1→2) - β-D-galactopyranoside (13), 8-epikingiside (14), 3, 5-dicaffeoylquinicacid (15), and 2-methylphenyl-1-O-β-D-glucopyranoside (16). Conclusion: All the compounds are isolated from S. davidi for the first time.


Zeng J.-Y.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Zeng J.-Y.,Huaihua University | Li S.-H.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Li S.-H.,Huaihua University | And 6 more authors.
Chinese Traditional and Herbal Drugs | Year: 2014

Objective: To observe the effect of monocrotaline (MCT) extract from Crotalaria sessiliflora on inducing human pancreatic cancer cell BxPC-3 into polyploid giant cells in vitro. Methods: BxPC-3 cells (1 × 104/mL) were inoculated with MCT (0, 5, 10, and 20 μg/mL) in RPMI-1640 for 72 h, respectively, then the cell morphology was observed by Giemsa staining, and the DNA content was measured by flow cytometry; BxPC-3 cells-induced apoptosis was checked by AnnexinV-FITC/PI double staining using flowcytometry, BxPC-3 polyploid giant cell genome was checked by chromosome spreading assay, and Cyclin B1 expression also was analysed by Western blotting. Results: Giemsa staining and DNA content by flow cytometry showed that MCT induced BxPC-3 cell into polyploid giant cells in vitro. MCT treatment for 72 h appeared 4N, and 8N polyploid giant of BxPC-3 cells, induction of apoptosis, decreased the expression of cyclin B1 in a dose dependent manner. Chromosome analysis demonstrated once again that polyploid giant cell was mainly in 8N. Conclusion: Monocrotaline might exert its antitumor effect by blocking the cell cycle, inducing apoptosis, and down-regulating Cyclin B1 protein.


Wei L.,Huaihua University | Wei L.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Wei L.,Key Laboratory of Hunan Higher Education for Hunan Western Medicinal Plant and Ethnobotany | Li S.,Huaihua University | And 17 more authors.
PLoS ONE | Year: 2014

Background: Houttuynia cordata Thunb. is an important traditional medical herb in China and other Asian countries, with high medicinal and economic value. However, a lack of available genomic information has become a limitation for research on this species. Thus, we carried out high-throughput transcriptomic sequencing of H. cordata to generate an enormous transcriptome sequence dataset for gene discovery and molecular marker development. Principal Findings: Illumina paired-end sequencing technology produced over 56 million sequencing reads from H. cordata mRNA. Subsequent de novo assembly yielded 63,954 unigenes, 39,982 (62.52%) and 26,122 (40.84%) of which had significant similarity to proteins in the NCBI nonredundant protein and Swiss-Prot databases (E-value <10-5), respectively. Of these annotated unigenes, 30,131 and 15,363 unigenes were assigned to gene ontology categories and clusters of orthologous groups, respectively. In addition, 24,434 (38.21%) unigenes were mapped onto 128 pathways using the KEGG pathway database and 17,964 (44.93%) unigenes showed homology to Vitis vinifera (Vitaceae) genes in BLASTx analysis. Furthermore, 4,800 cDNA SSRs were identified as potential molecular markers. Fifty primer pairs were randomly selected to detect polymorphism among 30 samples of H. cordata; 43 (86%) produced fragments of expected size, suggesting that the unigenes were suitable for specific primer design and of high quality, and the SSR marker could be widely used in markerassisted selection and molecular breeding of H. cordata in the future. Conclusions: This is the first application of Illumina paired-end sequencing technology to investigate the whole transcriptome of H. cordata and to assemble RNA-seq reads without a reference genome. These data should help researchers investigating the evolution and biological processes of this species. The SSR markers developed can be used for construction of high-resolution genetic linkage maps and for gene-based association analyses in H. cordata. This work will enable future functional genomic research and research into the distinctive active constituents of this genus. © 2014 Wei et al.


Ou L.,Huaihua University | Ou L.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Ou L.,Vegetable Institution of Hunan Academy of Agricultural Science | Ou L.,Key Laboratory of Hunan Higher Education for Hunan western Medicinal Plant and Ethnobotany | And 2 more authors.
Shengtai Xuebao/ Acta Ecologica Sinica | Year: 2012

Increased human activities have had a significant impact on the global climate. Abnormal changes, such as drought stress caused by long-term un-rain, can affect plant growth and development. Pepper (Solanaceae), originally planted in tropical areas of South America, not only has medicinal value it has the highest vitamin C content of all vegetables. The International Board for Plant Cenetic Resources (IBPCR) has classified pepper species into five species; Capsicum annuum L. (CA), Capsicum baccatum L. (CB), Capsicum chínense Jacquin. (CC), Capsicum frutescens L. (CF), Capsicum pubescens Ruiz and Pavon (CP). CA is the most differentiated, widely cultivated species being the focus of pepper breeding worldwide. Screening of resistant germplasms is one of the most important measures against abnormal climate changes. Pot experiments were employed to determine the photosynthetic response and associated physiological characters, such as superoxide dismutase (SOD) and glutathione reductase (CR) activities, root activity and adventitious root number, of five domesticated and one wild pepper species (C baccatum var. baccatum (CBY)) to drought stress. The photosynthetic pigment content decreased the net photosynthetic rate (PN) of all tested peppers continued to decline on the first day of regular irrigation, then started to increase on the second day, and almost recovered to normal levels by the seventh day. Non-photochemical quenching (q N), water use efficiency (WUE), and root activity had increased dramatically, the SOD and CR activities of all the pepper species first increased, being highest on day 4, and subsequently decreased. The number of adventitious roots slightly increased in all peppers, suggesting that pepper has a stronger resistance to drought stress. The results also show that changes in the above parameters vary in different species. CB had smaller decreases in photosynthetic pigment content, PN and stomatal conductance (gs,), and greater increases in qN, WUE, SOD and CR activities and root activity, indicating that CB has the strongest resistance to drought stress. CC and CF showed bigger decreases in photosynthetic pigment content, PN and gs with smaller increases in qN, WUE, SOD, and CR activities and root activity, indicating they have weaker drought stress resistance. In addition, the wild pepper, C. baccatum var. baccatum (CBY), had the smallest decrease in photosynthetic pigment content, PN and gs with the greatest increases in qN, WUE, SOD, and CR activities, and root activity. This indicates that CBY has better resistance to adverse environments than all tested species, which is perhaps caused by the loss of stress resistance genes during the process of domestication. Our study found that, of the species tested, the ability to resist drought stress is strongest in wild pepper CBY. Of the five species tested, CB had the highest photosynthetic rate and strongest drought resistance capability. In conclusion, we suggest that crossbreeding of domestic and wild pepper species be considered for commercial breeding. Such a measure will not only expand the genetic pool, increasing genetic distance and eventually increasing production; it will also to introduce elite stress tolerance genes improving each species' ability to resist adverse environments.


Ou L.-J.,Key Laboratory of Hunan Higher Education for Hunan western Medicinal Plant and Ethnobotany | Ou L.-J.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Ou L.-J.,Huaihua University | Zhang R.-W.,Huaihua University | And 5 more authors.
Chinese Traditional and Herbal Drugs | Year: 2011

Objective: The nuclear DNA ITS sequence variation and its correlation to geographical distribution of Asparagus cochinchinensis from different populations in China were studied, the aims were to reveal the difference among different populations and determine the genuineness of A. cochinchinensis. Methods: The ITS fragments of 75 samples from 25 populations were amplified and bidirectional sequenced. And they were analyzed using the software of DNAMAN and MEGA 4. Results: ITS sequences of three samples from the same areas were almost the same and ITS sequence of 25 populations were different from each other. The variation of ITS2 sequences was bigger than that of ITS1sequences. ITS sequences of Guizhou Province had bigger genetic differentiation and more variable sites and information sites than other provinces had. Clustering results based ITS sequences showed three samples from the same area firstly clustered and then populations from a province clustered. All populations were divided into two branches based on North latitude of 24.6°-36.6°and 22.2°-24.4°, the first branch included the populations of Qinghai, Hunan, and Guizhou Provinces, the second branch included the populations of Zhejiang, Guangdong, Yunnan and Guangxi Province. Conclusion: ITS sequences could identify the different populations of A. cochinchinensis. Guizhou Province is the place of genuineness of A. cochinchinensis. Clustering the different populations of A. cochinchinensis primarily relates to the latitude while has little relationship with the longitude.


Ou L.-J.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Ou L.-J.,Key Laboratory of Hunan Higher Education for Hunan western Medicinal Plant and Ethnobotany | Ou L.-J.,Huaihua University | Yan W.,Huaihua University | And 5 more authors.
Chinese Traditional and Herbal Drugs | Year: 2011

Objective: To establish and optimize ISSR-PCR reaction system for Asparagus cochinchinensis and lay foundation for its genetic diversity research. Methods: The single-factor test and orthogonal design were applied for optimizing seven factors in the ISSR-PCR reaction system including Mg2+, dNTP, primers, Taq DNA polymerase, the template DNA, extension time, and cycle times. Results: The suitable PCR reaction system contained 1.25 mmol/L Mg2+, 320 μmol/L dN TP, 1.2 μmol/L primer, 1.5 U Taq DNA polymerase, and 40 ng template DNA in total 25 μL reaction solution. On this basis, 13 primers were screened with stable amplification and rich polymorphism from 50 ISSR primers. The optimal annealing temperature for ISSR-PCR reaction was proposed by gradient PCR. Conclusion: It is proved that the established and optimized ISSR reaction system would be stable and credible by the germplasm testing result of 17 A. cochinchinensis populations. This would provide the basis for the genetic analysis of A. cochinchinensis.


Ou L.,Huaihua University | Ou L.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Ou L.,Key Laboratory of Hunan Higher Education for Hunan Western Medicinal Plant and Ethnobotany | Hu A.,Symbiosis Institute of Agricultural Engineering of Huaihua | And 2 more authors.
Nongye Gongcheng Xuebao/Transactions of the Chinese Society of Agricultural Engineering | Year: 2012

The photosynthetic characteristics, root characteristics and part of the agronomic traits of rice with the floating culture method (CW) and conventional tillage (CT) were studied. The results showed that photosynthetic rate (PN) and stomatal conductance (gs) in CW were higher than those in CT; compared with CT, rice cultivated in CW significantly enhanced root numbers, root activities and enzyme activities, and increased tillers and effective panicle number by 17.91% and 24.07%, respectively. No significant differences were observed in the water use efficiency (WUE), plant height, seed setting rate and the thousand seed mass between CW and CT. Rice with CW exhibited more root absorption, tillers and effective panicle number, and higher PN, thus achieve the purpose of the increase in production. This research suggests the floating culture method is worth popularizing.


Quan M.-H.,Huaihua College | Quan M.-H.,Key Laboratory of Hunan Higher Education for Hunan Western Medicinal Plant and Ethnobotany | Quan M.-H.,Key Laboratory of Hunan Province for Study and Utilization of Ethnic Medicinal Plant Resources | Ou L.-J.,Huaihua College | And 7 more authors.
Chinese Traditional and Herbal Drugs | Year: 2012

Objective: Establishing the rapid propagation for plant tissue culture of Asparagus cochinchinensis in order to rationally protect and utilize the medicinal plant resources of A. cochinchinensis. Methods: Different media and plant growth regulators at different concentration were used to A. cochinchinensis explants for callus induction, cluster buds induction, and rooting. Results: The best medium for callus induction was 1/2 MS + 1.0 mg/L 6-BA + 0.5 mg/L NAA with pH 5.8; The optimal medium for cluster buds induction was MS + 1.5 mg/L 6-BA + 0.5 mg/L IAA with pH 5.8; The optimal medium for rooting was 1/2 MS + 0.5 mg/L IBA + 0.5 mg/L IAA with pH 5.8 (iron salt in MS medium reduced to 6.95 mg/L and sucrose decreased to 20 g/L). Conclusion: Different explants could be induced for tissue culture and tender bud is the optimal tissue culture explants with the highest induction rate. An effective approach for rapid propagation has been provided, which could promote the artificial cultivation of A. cochinchinensis.

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