ABSTRACT
Decaying bamboo in freshwater is a unique eco-environment for fungi. Three new Distoseptispora (Distoseptisporaceae) species, D.meilingensis, D.yongxiuensis and D.yunjushanensis from submerged decaying bamboo culms in Jiangxi Province, China, were discovered, based on phylogenetic analyses and morphological characters. The combined data of ITS-LSU-SSU-Tef1 sequences were used to infer the phylogenetic relationship between D.meilingensis, D.yongxiuensis, D.yunjushanensis and related species. Both molecular analyses and morphological data supported D.meilingensis, D.yongxiuensis and D.yunjushanensis as three independent taxa.
ABSTRACT
Rhei Radix et Rhizoma is one of the most used medicinal materials in China. Its original species are Rheum palmatum, Rh. tanguticum, and Rh. officinale. Rhei Radix et Rhizoma derived from different original species are significantly different in their active ingredients and pharmacological effects. To develop an accurate, rapid, and specific identification method, we obtained the chloroplast genomes of the three original species by Illumina Novaseq sequencing. We designed specific DNA barcodes from the hypervariable regions, which can accurately identify the three original species. The experimental results showed that the total length of the chloroplast genomes of Rh. tanguticum, Rh. officinale and Rh. palmatum were 161 039 bp, 161 093 bp, and 161 136 bp, respectively. All the three genomes were represented as typical quadripartite structures. A total of 131 genes, including 86 protein-coding genes, 37 transfer RNA (tRNA) genes, and eight ribosomal RNA (rRNA) genes were identified from each chloroplast genome. Five pairs of primers based on the hypervariable regions were designed to efficiently amplify 42 samples. Results confirmed that five hypervariable regions, rps16-trnQ, psaA-ycf3, psbE-petL, ndhF-rpl32, and trnT-trnL, can be used as specific DNA barcodes for the identification of Rh. tanguticum, Rh. officinale, and Rh. palmatum. These results provided genetic information for further species identification of Rhei Radix et Rhizoma, and improve the safety of this clinical medication as well as standardize the market for Rhei Radix et Rhizoma.
ABSTRACT
Licorice, one of the most commonly used medicinal materials in China, grows mainly in arid and semi-arid regions and has important economic and ecological values. Basic leucine zipper (bZIP) transcription factors in plants play an important role in regulating biological or abiotic stress responses, growth, and secondary metabolite synthesis. bZIP transcription factors in the published whole genome database of Glycyrrhiza uralensis were identified using bZIP sequences found in Arabidopsis thaliana genome as reference, and ABA-dependent bZIP genes were identified by using Illumina high-throughput sequencing. The physical and chemical properties, structure of the encoded proteins, and the gene expression patterns with exogenous ABA stress were analyzed. A total of 69 bZIP transcription factor genes were identified in G. uralensis, named Gubzip1-69, and they were divided into 10 subfamilies (A-I and S) according to their similarity to bZIPs of A. thaliana. By calculating the relative expression levels of the 69 GubZIPs genes under different concentrations of exogenous ABA stress, genes that may be involved in the regulation of ABA signaling pathways were identified, namely GubZIP1, GubZIP5, GubZIP8, GubZIP30, GubZIP33 and GubZIP56. The results of expression pattern analysis of these GubZIPs genes under exogenous ABA stress showed that the expression pattern of GubZIPs genes changed significantly with 50 mg·L-1 ABA. The relative expression levels of these genes decreased 3 h after treatment, and gradually increased 6 h after treatment. Except for GubZIP8, the relative expression levels of these genes were significantly increased after 12 h. Further research on the function of bZIP transcription factors of G. uralensis and elucidating their regulatory mechanisms should be of interest and will provide a scientific basis for cultivating high-quality cultivars of G. uralensis through molecular breeding methods.
ABSTRACT
italic>Dendrobium moniliforme is an important source of Dendrobii Caulis and one of the main sources of authentic Fengdou. The complete chloroplast genome of D. moniliforme was sequenced using Illumina Hiseq technology and its gene map and genomic structure were analyzed. Then comparative and phylogenetic analysis of the complete chloroplast genomes of D. moniliforme and its related species were conducted. The chloroplast genome of D. moniliforme was 150 754 bp in length and had a typical quadripartite structure with a large single copy (LSC, 84 818 bp), a small single copy (SSC, 14 124 bp) and two inverted repeats (IRs, 25 906 bp each). A total of 123 chloroplast genes were annotated, including 77 protein-coding genes, 38 tRNA genes and 8 rRNA genes, of which 17 genes contained introns. Bioinformatics analysis identified 53 SSR sites, most of which had A-T base preference. A phylogenetic tree was constructed using the chloroplast genome sequences of 33 Dendrobium species. The results showed that Dendrobium complex species were clustered in a single large branch, indicating that they were closely related. This study provides a scientific basis for the identification of D. moniliforme and the phylogenetic relationship of D. moniliforme complex species necessary for Herbgenomics research.
ABSTRACT
As abscisic acid (ABA) receptor, the pyrabactin resistance 1-like (PYR/PYL) protein (named PYL for simplicity) plays an important part to unveil the signal transduction of ABA and its regulatory mechanisms. Glycyrrhiza uralensis, a drought-tolerant medicinal plant, is a good model for the mechanism analysis of ABA response and active compound biosynthesis. However, knowledge about PYL family in G. uralensis remains largely unknown. Here, 10 PYLs were identified in G. uralensis genome. Characterization analysis indicated that PYLs in G. uralensis (GuPYLs) are relatively conserved. Phylogenetic analysis showed that GuPYL1-3 belongs to subfamily I, GuPYL4-6 and GuPYL10 belong to subfamily II and GuPYL7-9 belongs to subfamily III. In addition, transcriptome data presented various expression levels of GuPYLs under different exogenous ABA stresses. The expression pattern of GuPYLs was verified by Quantitative real-time polymerase chain reaction (qRT-PCR). The study proved that GuPYL4, GuPYL5, GuPYL8 and GuPYL9 genes are significantly up-regulated by ABA stress and the response process is dynamic. This study paves the way for elucidating the regulation mechanism of ABA signal to secondary metabolites and improving the cultivation and quality of G. uralensis using agricultural strategies.
ABSTRACT
To investigate the effects of the expression of coumaroylquinate 3’-monooxygenase (C3’H) and shikimate O-hydroxycinnamoyltransferase (HCT) in the chlorogenic acid-producing pathway and active ingredients in Chrysanthemum morifolium under flooding stress, we cloned two C3’H genes which were CmC3’H1 and CmC3’H2 and two HCT genes which were CmHCT1 and CmHCT2 by the RT-PCR from Hangju and conducted bioinformatics analysis. During the flower bud differentiation stage, we flooded the C. morifolium and then used β-actin as the reference gene to detect the relative expression of the four genes by the qRT-PCR. Finally, the content of downstream products and other indicators of these four genes in C. morifolium were measured by HPLC. We obtained the four genes’ complete open reading frame and predicted the relative molecular mass of the amino acid sequence and the theoretical isoelectric point (pI). And the protein tertiary structure models were constructed. The qRT-PCR results showed that flooding the C. morifolium for 3 days during the flower bud differentiation stage resulted in significant expression changes of the four genes at different growth stages. The results of HPLC showed that chlorogenic acid, the downstream product catalyzed by the C3’H and the HCT, was significantly higher than that in the control group. It was also found that the content of luteoloside and 3,5-O-di-caffeoylquinic acid was also significantly higher than those of the control group. Therefore, C. morifolium regulates the synthesis of downstream products by regulating the expression of the four genes under flooding stress, thereby responding to flooding stress. And the flooding stress during flower bud differentiation can significantly enhance the accumulation of active ingredients of C. morifolium.
ABSTRACT
To investigate the effects of the expression of flavonoid 3' hydroxylase gene ( and active ingredients in under flooding stress, we cloned F3'H from Hangju (temporarily named ) and conducted bioinformatics analysis. During the flower bud differentiation stage, we flooded the and then used the Real-time PCR to detect the relative expression of ; Finally, active ingredients of the inflorescence were measured by HPLC.The sequencing results showed that 1 562 bp sequence was acquired with the largest open reading frame of 1 527 bp, which encoded 508 amino acids. The phylogenetic tree found that was highly homologous to other species of Compositae. Real-time PCR results showed that had a significant response to flooding stress and had the highest expression level after flooding for 24 h, which was about 9 times as that of the control group. The results of HPLC showed that luteolin and luteoloside, the downstream products catalyzed by the F3'H, were significantly higher than those in the control group. It was also found that the contents of chlorogenic acid and 3,5- acid were also significantly higher than those of the control group. Therefore, regulates the synthesis of downstream products by regulating the expression of in the flavonoid synthesis pathway under flooding stress, thereby responding to flooding stress. The flooding stress during flower bud differentiation can significantly enhance the accumulation of active ingredients.
Subject(s)
Chrysanthemum , Genetics , Cloning, Molecular , Cytochrome P-450 Enzyme System , Genetics , Floods , Gene Expression Regulation, Plant , Glucosides , Luteolin , Phylogeny , Plant Proteins , Genetics , Stress, PhysiologicalABSTRACT
BACKGROUND: Antioxidants have been shown to enhance the proliferation of adipose-derived mesenchymal stem cells (ADMSCs) in vitro, although the detailed mechanism(s) and potential side effects are not fully understood. RESULTS: During log-phase growth, exposure to ImF-A resulted in a higher percentage of ADMSCs in the S phase of the cell cycle and a smaller percentage in G0/G1 phase. This resulted in a significantly reduced cell-doubling time and increased number of cells in the antioxidant-supplemented cultures compared with those supplemented with FGF-2 alone, an approximately 225% higher cell density after 7 days. Western blotting showed that the levels of the CDK inhibitors p21 and p27 decreased after ImF-A treatment, whereas CDK2, CDK4, and CDC2 levels clearly increased. In addition, ImF-A resulted in significant reduction in the expression of CD29, CD90, and CD105, whereas relative telomere length, osteogenesis, adipogenesis, and chondrogenesis were enhanced. The results were similar for ADMSCs treated with antioxidants and those under hypoxic conditions. CONCLUSION: Antioxidant treatment promotes entry of ADMSCs into the S phase by suppressing cyclin-dependent kinase inhibitors and results in rapid cell proliferation similar to that observed under hypoxic conditions.
Subject(s)
Adipose Tissue/cytology , Antioxidants/administration & dosage , Cell Proliferation/drug effects , Mesenchymal Stem Cells/drug effects , CDC2 Protein Kinase , Cell Division/drug effects , Cell Hypoxia/drug effects , Cell Line , Cyclin-Dependent Kinase 4/antagonists & inhibitors , Cyclin-Dependent Kinase 4/biosynthesis , Cyclin-Dependent Kinases/antagonists & inhibitors , Cyclin-Dependent Kinases/biosynthesis , Fibroblast Growth Factor 2/biosynthesis , Gene Expression Regulation/drug effects , Humans , Mesenchymal Stem Cells/cytologyABSTRACT
OBJECTIVE: To study the roles of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA in peripheral blood as well as serum total immunoglobulin E (IgE) in the pathogenesis of bronchiolitis caused by respiratory syncytial virus (RSV). METHODS: The proportion of CD4(+)CD25(+) regulatory T cells and expression of Foxp3 mRNA in peripheral blood, and total serum IgE level were tested by flow cytometry, RT-PCR and ELISA respectively in 57 children with RSV bronchiolitis (26 atopic patients and 31 nonatopic patients). Twenty five healthy children were used as the control group. RESULTS: The proportion of CD4(+)CD25(+) regulatory T cells in peripheral blood in children with bronchiolitis, either in the atopic (7.7+/- 1.6%)or the nonatopic group (8.8+/- 2.1%), was significantly lower than that in the control group (10.5+/- 1.6%) (P< 0.01). Foxp3 mRNA expression in peripheral blood was significantly lower in both atopic and nonatopic children with bronchiolitis than that in the control group (P< 0.01). Significantly increased total serum IgE level was noted in both atopic (241.2+/- 102.5 IU/mL) and nonatopic children (125.5+/- 63.2 IU/mL) with bronchiolitis compared with that in the control group (27.2+/- 10.5 IU/ml) (P< 0.01). There were significant differences in the proportion of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA expression in peripheral blood (P< 0.05) as well as total serum IgE level (P< 0.01) between the atopic and the nonatopic group. The proportion of CD4(+)CD25(+) regulatory T cells (r=-0.70, P< 0.01) and Foxp3 mRNA expression in peripheral blood (r=-0.79, P< 0.01) were closely negatively correlated to total serum IgE level. CONCLUSIONS: Both the proportion of CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA expression in peripheral blood were reduced, in contrast, the total serum IgE level increased in children with RSV bronchiolitis. This suggested that CD4(+)CD25(+) regulatory T cells and Foxp3 mRNA together with IgE participated in the pathogenesis of RSV bronchiolitis.
Subject(s)
Bronchiolitis/immunology , Forkhead Transcription Factors/genetics , Immunoglobulin E/blood , RNA, Messenger/blood , Respiratory Syncytial Virus Infections/immunology , T-Lymphocytes, Regulatory/physiology , Bronchiolitis/etiology , CD4-Positive T-Lymphocytes/immunology , Female , Humans , Infant , Male , Respiratory Syncytial Virus Infections/etiologyABSTRACT
<p><b>OBJECTIVE</b>To study the ITS sequence variation of Pinellia ternata from different population in China, and it correlation to geographical distribution and morpha of the plant.</p><p><b>METHOD</b>The rDNA ITS regions of various P. ternata were amplified and sequenced. And they were analyzed by means of the software of CLUSTRAL and MEGA.</p><p><b>RESULT</b>Complete sequence of ITS and 5.8S rDNA of 16 different P. ternata population were obtained. The sequences of ITS1, 5.8S and ITS2 are 276,162 and 246 bp, respectively. ITS1 was more conservative than ITS2. Phylogenetic tree based on ITS1 and ITS2 sequences data was conducted by Neighbor-joining method.</p><p><b>CONCLUSION</b>Ribosomal DNA ITS sequence analyses can be applied to the resource research of P. ternata.</p>
