Abortifacient Agents, Nonsteroidal ; pharmacology ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Female ; Pinellia ; chemistry ; Plant Proteins ; pharmacology ; Pregnancy

In order to express PebC1 in Pichia pastoris, the pebC1 sequence was amplified from genome Botrytis cinerea BC-4-2-2-1 by PCR and subcloned into the Pichua pastoris expression vector pPIC9K to generate pPIC9K-pebC1. The recombinant plasmid was linearized by Bgl II and transformed into Pichia pastoris GS115 by electroporation. Recombinant Pichia pastoris GS115/pPIC9K-pebC1 was screened by MD and G418-YPD plates and further confirmed by PCR. The protein expression was induced by methanol and analyzed by SDS-PAGE. SDS-PAGE analysis showed a special band about 39 kDa and western blotting indicated a good antigenicity of the expressed protein. Bioassay results showed that the recombinant protein PebC1 can induce resistance to gray mould disease of cucumber and Arabidopsi thaliana.
Botrytis ; chemistry ; Fungal Proteins ; biosynthesis ; genetics ; pharmacology ; Pichia ; genetics ; metabolism ; Plant Diseases ; prevention & control ; Recombinant Proteins ; biosynthesis ; genetics ; pharmacology

As one of the important active components of traditional Chinese medicine (TCM), plant origin active proteins have many significant pharmacological functions. According to researches on the plant origin active proteins reported in recent years, pharmacological effects include anti-tumor, immune regulation, anti-oxidant, anti-pathogeny microorganism, anti-thrombus, as well as hypolipidemic and hypoglycemic activities of plant origin were reviewed, respectively. On the other hand, the analytical methods including chromatography, spectroscopy, electrophoresis and mass spectrometry for plant origin proteins analysis were also summarized. The main purpose of this paper is providing a reference for future development and application of plant active proteins.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Antioxidants ; pharmacology ; Fibrinolytic Agents ; pharmacology ; Humans ; Hypoglycemic Agents ; pharmacology ; Immunologic Factors ; pharmacology ; Plant Proteins ; analysis ; pharmacology ; Research

The NPRI (nonexpressor of pathogenesis-related genes (1) gene, firstly cloned in Arabidopsis thaliana, is a key gene involved in regulation of plant disease resistance. It plays a pivotal role not only in systemic acquired resistance (SAR) and induced systemic resistance (ISR), but also in basic resistance and resistance (R) gene-dependent resistance. NPR1 monomerization induced by enhanced reducing condition after oxidative burst, and the accumulation of NPR1 monomers in the nuclei, are required and enough for expression of PR (pathogenesis-related) genes and SAR. NPR1 regulates PR gene expression through interaction with TGA transcription factors (TF). As a cross-talk point of a variety of defense signaling pathways, probably through direct or indirect interacting with some WRKY TFs and a NPR1-like protein NPR4, NPR1 is essential in balancing salicylic acid- and jasmonic acid- dependent signal transduction pathways, which is achieved through an unknown mechanism in the cytosol. The possible application of NPR1 in plant protection is also discussed in this review.
Arabidopsis Proteins ; genetics ; Cyclopentanes ; metabolism ; pharmacology ; Gene Expression Regulation, Plant ; genetics ; Oxylipins ; metabolism ; pharmacology ; Plant Diseases ; genetics ; Plants, Genetically Modified ; Salicylic Acid ; metabolism ; pharmacology ; Signal Transduction

OBJECTIVETo study the proteomics of Pinellia pedatisecta Schott total protein on human ovarian cancer SKOV3 cells.

METHODSSKOV3 cells were in vitro cultured. The growth inhibition of SKOV3 cells in the logarithmic phase with different concentrations of Pinellia pedatisecta Schott (0, 0.05, 0.10, 0.20, 0.30, 0.40, and 0.50 mg/mL) at different time points (24, 48, 72, and 96 h) was analyzed by CCK-8 colorimetry. The total protein was extracted after adding 0.296 mg/mL Pinellia pedatisecta Schott protein for 48 h. The protein with differential expressions was screened out using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS).

RESULTSThe protein of Pinellia pedatisecta Schott (at 0.10 - 0.50 mg/mL) could significantly inhibit the growth of SKOV3 cells in a time- and dose-dependent manner (P < 0.05, P < 0.01). After analyzed by 2-DE and MALDI-TOF-MS, 43 differential protein dots were successfully identified (21 up-regulated and 22 down-regulated), including alpha-enolase 1, eukaryotic initiation factor 3alpha, cyclophilin B, and so on.

CONCLUSIONSProtein of Pinellia pedatisecta Schott could significantly inhibit the growth of SKOV3 cells, and lead to the proteomics changes of SKOV3 cell strain. They might be correlated with its anti-tumor mechanisms.

Cell Line, Tumor ; Female ; Humans ; Ovarian Neoplasms ; metabolism ; Pinellia ; Plant Proteins ; pharmacology ; Proteomics ; methods

Plant adaptation to adverse environment depends on transmitting the external stress signals into internal signaling pathways, and thus forming a variety of stress response mechanisms during evolution. Brassinosteroids (BRs) is a steroid hormone and widely involved in plant growth, development and stress response. BR is perceived by cell surface receptors, including the receptor brassinosteroid-insensitive 1 (BRI1) and the co-receptor BRI1-associated-kinase 1 (BAK1), which in turn trigger a signaling cascade that leads to the inhibition of BIN2 and activation of BES1/BZR1 transcription factors. BES1/BZR1 can directly regulate the expression of thousands of downstream responsive genes. Studies in the model plant Arabidopsis thaliana have shown that members of BR biosynthesis and signal transduction pathways, particularly protein kinase BIN2 and its downstream transcription factors BES1/BZR1, can be extensively regulated by a variety of environmental factors. In this paper, we summarize recent progresses on how BR biosynthesis and signal transduction are regulated by complex environmental factors, as well as how BR and environmental factors co-regulate crop agronomic traits, cold and salt stress responses.
Arabidopsis/metabolism* ; Brassinosteroids/pharmacology* ; DNA-Binding Proteins/metabolism* ; Gene Expression Regulation, Plant ; Stress, Physiological

OBJECTIVETo investigate anti-attachment effect of ethyl acetate extract of Huanglian Jiedu decoction (EAHD) on Candida glabrata.

METHODSerial 2-fold dilution assay was used to determine the minimum inhibitory concentrations MICs of EAHD to C. glabrata. XTT assay was used to evaluate the effect of EAHD against adhesion of C. glabrata. Inverted microscope, scanning electron microscope (SEM) and fluorescein diacetate (FDA) staining were applied to observe the morphological changes of C. glabrata in adhesion. PCR was adopted to inspect the expression of attachment-related genes such as EPA1, EPA6 and EPA7.

RESULTThe MIC of EAHD and fluconazole to C. glabrata were 320 mg · L(-1) and 1 mg · L(-1) respectively. The total cells including budding cells decreased in a dose-dependent manner following EAHD treatment. The expressions of EPA1, EPA6 and EPA7 were downregulated dramatically after EAHD treatment.

CONCLUSIONEAHD could effectively inhibit adherence of C. glabrata.

Acetates ; Candida glabrata ; drug effects ; physiology ; Drugs, Chinese Herbal ; pharmacology ; Fungal Proteins ; genetics ; Lectins ; genetics ; Plant Extracts ; pharmacology

In order to investigate the effects of fungal elicitor and macroporous adsorption resin on shikonin accumulation in hairy roots of arnebia euchroma (Royle) Johnst, we used spectrophotometry to determine the total naphthoquinone content of the hairy roots, by adding different volume ratio of Aspergillus niger elicitor, Aspergillus oryzae elicitor, and the macroporous resin into the M-9 liquid medium at different culture time. The results show that the total naphthoquinone content was 2.28 times higher than the control when we added mixed elicitors of Aspergillus niger and Aspergillus oryzae at the ratio of 2.5:50 in the 10th day of hairy roots cultivating. The total naphthoquinone content was 3.71 times higher than that of the control, when we added macroporous adsorption resin NKA-9. Aspergillus niger elicitor exhibited synergistic effect with Aspergillus oryzae elicitor to enhance the naphthoquinone. Also, the total naphthoquinone level was 4.17 times higher than that of the control by adding mixed fungal elicitor and macroporous adsorption resin NKA-9 in the bioreactor. Aspergillus oryzae and mixed elicitor could promote the hairy roots proliferation, and macroporous adsorption resin NKA-9 and mixed elicitor increased the total naphthoquinone content. In summary, the measure developed for Arnebia euchroma (Royle) Johnst hairy roots cultivating in bioreactors may potential for large-scale production of naphthoquinone.
Aspergillus niger ; metabolism ; Boraginaceae ; metabolism ; Fungal Proteins ; pharmacology ; Naphthoquinones ; analysis ; metabolism ; Plant Roots ; chemistry ; metabolism ; Porosity ; Resins, Synthetic ; pharmacology

OBJECTIVE: This study explored the rejuvenation mechanisms of Thai polyherbal medicines using different approaches, including in vitro methods, as well as a well-defined nematode model, Caenorhabditis elegans. METHODS: THP-R-SR012 decoction was selected from 23 polyherbal medicines, based on metal-chelating and chain-breaking antioxidant capacities. The influences of this extract on the survival and some stress biomarkers of C. elegans under paraquat-induced oxidative stress were evaluated. Furthermore, lifespan analysis and levels of lipofuscin accumulation were examined in senescent nematodes. The phytochemical profile of THP-R-SR012 was analyzed. RESULTS: Supplementation with THP-R-SR012 decoction significantly increased the mean lifespan and reduced the oxidative damage to C. elegans under oxidative stress conditions. Further, THP-R-SR012 supplementation slightly influenced the lifespan and the level of lipofuscin accumulation during adulthood. Antioxidant-related phytochemical constituents of THP-R-SR012 decoction were rutin, naringenin, 3,4-dihydroxybenzoic acid, gallic acid, glycyrrhizic acid, demethoxycurcumin and 18α-glycyrrhetinic acid. CONCLUSION The antioxidant potential of THP-R-SR012 was due to its scavenging properties, its enhancement of antioxidant-related enzyme activities, and the presence of the antioxidant-related compound. These results support the traditional use of THP-R-SR012 decoction as a tonic for nourishing and strengthening the whole body.
Animals ; Antioxidants/pharmacology* ; Caenorhabditis elegans/metabolism* ; Caenorhabditis elegans Proteins/metabolism* ; Oxidative Stress ; Plant Extracts/pharmacology* ; Reactive Oxygen Species ; Rejuvenation ; Thailand

14-3-3 proteins are important proteins in plants, as they regulate plant growth and development and the response to biotic or abiotic stresses. In this study, a 14-3-3 gene(GenBank accession: OM683281) was screened from the cDNA library of the medicinal species Salvia miltiorrhiza by yeast two-hybrid and cloned. The open reading frame(ORF) was 780 bp, encoding 259 amino a cids. Bioinformatics analysis predicted that the protein was a non-transmembrane protein with the molecular formula of C_(1287)H_(2046)N_(346)O_(422)S_9, relative molecular weight of 29.4 kDa, and no signal peptide. Homologous sequence alignment and phylogenetic tree analysis proved that the protein belonged to 14-3-3 family and had close genetic relationship with the 14-3-3 proteins from Arabidopsis thaliana, Oryza sativa, and Nicotiana tabacum. The 14-3-3 gene was ligated to the prokaryotic expression vector pGEX-4 T-1 and then transformed into Escherichia coli BL21 for the expression of recombinant protein. Real-time fluorescent quantitative PCR showed that the expression of this gene was different among roots, stems, leaves, and flowers of S. miltiorrhiza. To be specific, the highest expression was found in leaves, followed by stems, and the lowest expression was detected in flowers. S. miltiorrhiza plants were treated with 15% PEG(simulation of drought), and hormones salicylic acid, methyl jasmonate, and ethephon, respectively, and the expression of 14-3-3 gene peaked at the early stage of induction. Therefore, the gene can quickly respond to abiotic stresses such as drought and plant hormone treatments such as salicylic acid, jasmonic acid, and ethylene. This study lays the foundation for revealing the molecular mechanism of 14-3-3 protein regulating tanshinone biosynthesis and responding to biotic and abiotic stresses.
14-3-3 Proteins/metabolism* ; Amino Acid Sequence ; Cloning, Molecular ; Ethylenes/metabolism* ; Gene Expression Regulation, Plant ; Hormones/metabolism* ; Phylogeny ; Plant Growth Regulators/pharmacology* ; Plant Proteins/metabolism* ; Recombinant Proteins/genetics* ; Salicylic Acid/metabolism* ; Salvia miltiorrhiza/metabolism*