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1.
Article in Chinese | WPRIM | ID: wpr-888069

ABSTRACT

Sanguinarine is the main active component of the Papaver plants, and protopine-6-hydroxylase(P6 H), involved in the sanguinarine biosynthetic pathway, can oxidize protopine to 6-hydroxyprotopine. The investigation on the diversity of P6 H genes in the medicinal Papaver plants contributes to the acquirement of P6 H with high activity to increase the biosynthesis of sanguinarine. Five P6 H genes in P. somniferum, P. orientale, and P. rhoeas were discovered based on the re-sequencing data of the Papaver species, followed by bioinformatics analysis. With the elongation factor 1α(EF-1α), which exhibits stable expression in the root and stem, as the internal reference gene, the transcription levels of P6H genes in roots and stems of the Papaver plants were detected by real-time fluorescent quantitative PCR. As indicated by the re-sequencing results, there were two genotypes of P6H in P. somniferum and P. orientale, respectively, and only one in P. rhoeas. The bioinformatics analysis showed that the P6 H proteins of the three Papaver plants contained the conserved domain cl12078, which is the characteristic of p450 supergene family, and transmembrane regions. The existence of signal peptide remained verification. Real-time fluorescent quantitative PCR results revealed that the transcription level of P6 H in roots of P. somniferum was about 1.44 times of that in stems(α=0.05). The present study confirmed genetic diversity of P6 H in the three medicinal Papaver plants, which lays a basis for the research on the biosynthesis pathway and mechanism of sanguinarine in Papaver species.


Subject(s)
Benzophenanthridines , Berberine Alkaloids , Cytochrome P-450 Enzyme System/genetics , Genetic Variation , Papaver/genetics
2.
Article in Chinese | WPRIM | ID: wpr-906404

ABSTRACT

Objective:To screen out the suitable nonpolar molecular cosolvent and concentration with adventitious root phenotype and ginsenoside content in the controlled experiment as the evaluation indexes, so as to lay a solid foundation for exploring the causes for good shape and high quality of <italic>Panax quinquefolium</italic>. Method:After being treated with different concentrations of dimethyl sulfoxide (DMSO) and ethanol, the adventitious roots were scanned using a panoramic scanner, and the resulting images were used for measuring the branch number and average diameter by WinRHIZO Pro 2016, Synbiosis ProtoCol 3 colony counter, Image J, and SmartRoot. The contents of ginsenosides Rg<sub>1</sub>, Rb<sub>1</sub>, and Re were determined by ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Result:Compared with the blank control, the 0.1% DMSO and 75% ethanol made the adventitious root phenotype and ginsenoside contents significantly changed. Specifically, the branch number and average diameter were significantly reduced. The ginsenoside Rg<sub>1</sub> in the adventitious roots decreased after 0.1% DMSO treatment, whereas the ginsenosides Rg<sub>1</sub> and Re increased after 75% ethanol treatment. The adventitious root phenotype and ginsenoside contents in the 0.1% DMSO treatment group were not significantly different from those in the control group. Conclusion:The 0.01% DMSO does not affect the adventitious root growth of <italic>P. quinquefolium </italic>and is insoluble in water, enabling it to be considered as a suitable nonpolar molecular cosolvent for future research on the genetic causes for the good shape and high quality of <italic>P. quinquefolium</italic>.

3.
Article in Chinese | WPRIM | ID: wpr-906369

ABSTRACT

Objective:To explore the feasibility of replacing wood (or wood chips) with crop residues for culturing <italic>Armillaria gallica</italic> targeting the problems of forest resource destruction and increased cultivation cost caused by the extensive use of wood in <italic>Gastrodia elata</italic> cultivation, so as to reduce the cultivation cost of <italic>G. elata</italic>, promote the effective use of crop residues, and protect forest resources. Method:The growth situation of <italic>A. gallica</italic> in different media was observed, followed by the measurement of its growth rate using streaking method and the determination of total polysaccharide content of <italic>A. gallica</italic> by phenol-concentrated sulfuric acid colorimetric method. In order to further optimize the soybean straw cultivation medium, we carried out a four-factor three-level L<sub>9</sub>(3<sup>4</sup>) orthogonal assay on the ratio of main ingredients, sucrose content, inorganic salt content, and water content. Result:The comparison of growing states of <italic>A. gallica</italic> cultured in different media revealed that <italic>A. gallica</italic> in soybean straw medium began to grow since the fourth day of inoculation, and the mycelium grew well, with the growth rate being 0.352 cm·d<sup>-1</sup>, which was 1.48 times that in birch wood medium. The total polysaccharide content of <italic>A. gallica</italic> cultured in soybean straw medium was the highest, which was 39.260 mg·g<sup>-1</sup>, much higher than 17.028 mg·g<sup>-1</sup> of that cultured in birch wood medium. This demonstrated the obvious advantage of soybean straw medium, whose main ingredients were soybean straw and wheat bran at the ratio of 8:2, with the sucrose and inorganic salt content accounting for 1% and 0.5% of the main ingredients, respectively. When the water content reached 50%, the growth rate of <italic>A. gallica</italic> was maintained at 0.392 cm·d<sup>-1</sup>. Conclusion:This study has provided a basis for utilizing soybean straw instead of wood (or wood chips) as cultivation medium for <italic>A. gallica</italic>, thus better reducing the waste of forest resources and protecting the natural environment in the cultivation of <italic>G. elata</italic>.

4.
Article in Chinese | WPRIM | ID: wpr-906340

ABSTRACT

Objective:To explore the effects of diverse exogenous substances at different concentrations on the growth of<italic> Polyporus umbellatus</italic> mycelium and polysaccharide content and screen out the optimal growth condition for <italic>P. umbellatus</italic> mycelium, so as to provide a reference for its large-scale artificial cultivation. Method:<italic>P. umbellatus</italic> mycelium was cultured in media containing different exogenous substances using the method for fungal culturing in plate. The growth rate of the mycelium was judged by the colony diameter and the polysaccharide content was determined by the phenol-sulfuric acid method. Result:The high-dose cyclic adenosine monophosphate, 6-benzyl aminopurine (6-BA), gibberellic acid (GA), 2,4-dichlorophenoxyacetic acid (2,4-D), vitamin (V) B<sub>1</sub>, VB<sub>3</sub>, VB<sub>6</sub>, VB<sub>9</sub>, and VB<sub>12</sub> all promoted the growth of <italic>P. umbellatus</italic> mycelium and elevated polysaccharides content. By contrast, indole acetic acid (IAA), VC, and VB<sub>2</sub> inhibited its growth, with the most obvious inhibition detected in the high-dose VC group. IAA and VB<sub>2</sub> both reduced the polysaccharide content, whereas the high-dose VC significantly increased the polysaccharide content. Cyclic adenosine monophosphate, 6-BA, GA, 2,4-D, VB<sub>1</sub>, VB<sub>3</sub>, VB<sub>6</sub>, VB<sub>9</sub>, and VB<sub>12</sub> at the concentrations of 2 mmol·L<sup>-1</sup>, 6 mg·L<sup>-1</sup>, 15 mg·L<sup>-1</sup>, 2 mg·L<sup>-1</sup>, 4 mg·L<sup>-1</sup>, 2 mg·L<sup>-1</sup>, 4 mg·L<sup>-1</sup>, 6 mg·L<sup>-1</sup>, and 10 mg·L<sup>-1</sup>, respectively, contributed to the growth of <italic>P. umbellatus</italic> mycelium<italic> </italic>and polysaccharide accumulation. Conclusion:The growth of <italic>P. umbellatus </italic>mycelium and polysaccharide accumulation can be regulated by adding exogenous substances to the culture medium.

5.
Article in Chinese | WPRIM | ID: wpr-905971

ABSTRACT

Objective:To carry out germplasm resource evaluation and new variety breeding of <italic>Murraya paniculata</italic> and improve the germplasm quality, so as to ensure the demand, yield and quality of medicinal materials. Method:Following resource investigation and collection, 17 traits of 107 <italic>M. paniculata</italic> germplasm samples, like plant type, basal diameter, leaf shape, leaf length, and leaf width were determined and then subjected to principal component analysis and factor analysis for screening the principal component factors. Nine primary traits were selected as variables for further cluster analysis using Ward's method and Euclidean distance. According to the characteristics of medicinal parts, the core germplasms were screened out. Then the contents of auxin, zeatin, zeatin nucleoside, isopentenyl adenine, isopentenyl adenine riboside, dihydrozeatin, and dihydrozeatinriboside in the leaves were measured by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), followed by their correlation analysis with agronomic trait. Result:The variation coefficients of petiole length, branching number, and basal diameter were large. The nine main factors could be classified into four categories, with a contribution rate of 72.822%. The cluster analysis with Ward's method and Euclidean distance showed that 107 germplasm samples were clustered into six clusters and 61 core germplasms were identified. Such traits as leaf length, leaf width, petiole length, leaf surface, and petiole color were found to play an important role in the classification of <italic>M. paniculata</italic> germplasms. The content of zeatin nucleoside exhibited significant positive correlations with leaf length (<italic>P</italic><0.01), petiole length (<italic>P</italic><0.01), and leaf width (<italic>P</italic><0.05). Conclusion:These results have laid the foundation for further selection and breeding of <italic>M. paniculata</italic> new varieties.

6.
Article in Chinese | WPRIM | ID: wpr-921698

ABSTRACT

Schisandra sphenanthera is dioecious and only the fruits of female plants can be used as medicine and food. It is of great significance for the cultivation and production of S. sphenanthera to explore the differences between male and female plants at the non-flowering stage and develop the identification markers at non-flowering or seedling stage. In this study, the transcriptome of male and female leaves of S. sphenanthera at the non-flowering stage was sequenced by Illumina high-throughput sequencing technology and analyzed based on bioinformatics. A total of 236 682 transcripts were assembled by Trinity software and 171 588 were chosen as unigenes. Finally, 1 525 differentially expressed genes(DEGs) were identified, with 458 up-regulated and 1 067 down-regulated in female lea-ves. The down-regulated genes mainly involve photosynthesis, photosynthesis-antenna protein, carbon fixation in photosynthetic or-ganisms, and other pathways. Real-time quantitative PCR(qPCR) identified two genes between male and female leaves and one of them was a HVA22-like gene related to floral organ development and abscisic acid(ABA). Enzyme linked immunosorbent assay(ELISA) was applied to determine the content of ABA, auxin, gibberellin, and zeatin riboside(ZR) in leaves of S. sphenanthera. The results showed that the content of ABA and ZR in male leaves was significantly higher than that in female leaves. The involvement of down-regulated genes in female leaves in the photosynthesis pathway and the significant differences in the content of endogenous hormones between male and female leaves lay a scientific basis for analyzing the factors affecting sex differentiation of S. sphenanthera.


Subject(s)
Abscisic Acid , Gene Expression Profiling , Gene Expression Regulation, Plant , Plant Leaves/genetics , RNA-Seq , Schisandra , Transcriptome
7.
Article in Chinese | WPRIM | ID: wpr-888106

ABSTRACT

The longevity mechanism of ginseng(Panax ginseng) is related to its strong meristematic ability. In this paper, this study used bioinformatic methods to identify the members of the ginseng TCP gene family in the whole genome and analyzed their sequence characteristics. Then, quantitative real-time fluorescent PCR was performed to analyze the TCP genes containing elements rela-ted to meristem expression in the taproots, fibrous roots, stems, and leaves. According to the data, this study further explored the expression specificity of TCP genes in ginseng tissues, which facilitated the dissection of the longevity mechanism of ginseng. The ginseng TCP members were identified and analyzed using PlantTFDB, ExPASy, MEME, PLANTCARE, TBtools, MEGA and DNAMAN. The results demonstrated that there were 60 TCP gene family members in ginseng, and they could be divided into two classes: Class Ⅰ and Class Ⅱ, in which the Class Ⅱ possessed two subclasses: CYC-TCP and CIN-TCP. The deduced TCP proteins in ginseng had the length of 128-793 aa, the isoelectric point of 4.49-9.84 and the relative molecular mass of 14.2-89.3 kDa. They all contained the basic helix-loop-helix(bHLH) domain. There are a variety of stress response-related cis-acting elements in the promoter regions of ginseng TCP genes, and PgTCP20-PgTCP24 contained the elements associated with meristematic expression. The transcription levels of PgTCP20-PgTCP24 were high in fibrous roots and leaves, but low in stems, indicating the tissue-specific expression of ginseng TCP genes. The Class Ⅰ TCP members which contained PgTCP20-PgTCP23, may be important regulators for the growth and development of ginseng roots.


Subject(s)
Computational Biology , Gene Expression Regulation, Plant , Multigene Family , Panax/metabolism , Phylogeny , Plant Proteins/metabolism , Transcription Factors/metabolism
8.
Article in Chinese | WPRIM | ID: wpr-872854

ABSTRACT

Objiective: In the process of microRNA expression analysis by quantitative Real-time polymerase chain reaction(Real-time PCR),the selection of miRNA plays an important role in data standardization. Method:In this paper,13 Armillaria gallica.Candidate miRNAs were selected for bioinformatics analysis of their precursors,and the PMRD was used to predict similar sequences of their precursors,and the RNAfold was used to predict the secondary structure of the candidate miRNAs and their similar sequences. Real-time PCR was used to detect miRNAs expression in two genotypes of Armillaria gallica(genotype A,genotype B) before and after salt stress,and geNorm,NormFinder and BestKeeper were used to analyze the stability of miRNAs expression. Result:Secondary structure prediction and characterization of 9 candidate miRNA precursors showed that the miRNA predicted belonged to the miR family with typical stem-loop structure and the mature miRNAs were at the 5' or 3' end of the miRNA precursors.geNorm analysis showed that genotype A Armillaria gallica could select Novel-4* and Novel-9 as reference gene,genotype B could select Novel-9 and Novel-16 as its reference gene.NormFinder analysis showed that Novel-9 was stable in both genotype A and B Armillaria gallica.BestKeeper analysis showed that Novel-12* was stable in genotype A Armillaria gallica and Novel-2* was stable in genotype B Armillaria gallica. Conclusion:miRNA Novel-9 is the best stable reference gene,which lays a foundation for further research on the regulation mechanism of miRNA in Armillaria gallica.

9.
Article in Chinese | WPRIM | ID: wpr-872849

ABSTRACT

Endophytic bacteria exist in the host and do not cause obvious damage to the host,which is an important part of the host ecosystem. Endophytic bacteria are a kind of important microbial resources with diverse species and wide distribution. In the process of long-term coevolution between endophytic bacteria and host,many symbiotic relationships,such as antagonism and reciprocity,have been gradually formed,which can directly or indirectly regulate the growth and development of host,assist host to resist diseases and insect pests,environmental stress and other biological processes. Endophytic bacteria can produce a variety of secondary metabolites in the process of symbiosis with the host,which plays an important role in the development of new natural products. In recent years,endophytic bacteria and their secondary metabolites have been widely used in the research and development of new drugs,biological control and other fields. In this paper,the diversity,species identification and secondary metabolites of endophytic bacterial resources are reviewed,and the future development direction is discussed,hoping to provide reference for the further development and utilization of endophytic bacterial resources.

10.
Article in Chinese | WPRIM | ID: wpr-774577

ABSTRACT

Armillaria gallica is a facultative parasitic fungus which is the only nutrient source of Gastrodia elata during its cultivation.Chitinase,as a glycosidic hydrolytic enzyme,plays an important role in the growth,development,stress tolerance and symbiotic signal transduction of A. gallica. There were 22 chitinase genes in A. gallica. Bioinformatics analysis of amino acid sequence of these chitinase genes revealed that 12 chitinase genes contained glycosidase 18 family( GH18) domain. Chitinase amino acid sequences of A. gallica,A. ostoyae,G. elata,Saccharomyces cerevisiae and Trichoderma harzianum were analyzed byclustering trees,so as to further predict the gene function of chitinase in A. gallica. Induction of A. gallica branching with strigolactone analogue GR24,high-throughput sequencing technology based on the induction of branch group( MHJ1),uninduced branch group( MHJ2) and blank control group( MHJ3) is used to detect the expression quantity,the transcription level data of 22 chitinase genes were obtained and the heat map was generated for expression pattern analysis. It was found that 8 genes may be involved in physiological processes such as A. gallica branching,cell wall degradation and remodeling. In this paper,the function of chitinase gene in A. gallica was just preliminarily analyzed and predicted.


Subject(s)
Amino Acid Sequence , Armillaria , Chitinases , Computational Biology , Trichoderma
11.
Article in Chinese | WPRIM | ID: wpr-773674

ABSTRACT

The type and frequency of simple sequence repeats( SSRs) in the genomes was investigated using the DNA sequence data of Pueraria lobata and P. thomsonii. Based on these SSRs,20 pairs of SSR primers were designed and 5 high polymorphism primer pairs were selected to analyze genetic diversity of 9 cultivars of P. thomsonii in Jiangxi province. The results showed that the 5 pairs of primers could generate 16 polymorphic alleles bands. The average polymorphism information content( PIC) of each SSR primer pair was 0. 600 7.According to the genetic similarity coefficients,the 9 cultivars of P. thomsonii can be classified into 6 germplasms. This study established DNA identity cards with 5 pairs of SSR primers for different germplasm resources of P. thomsonii in Jiangxi province,which provided reference information for the selection of fine germplasms of P. thomsonii and the theoretical basis for the study of Dao-di herbs.


Subject(s)
China , DNA, Plant , Genetics , Genomics , Microsatellite Repeats , Polymorphism, Genetic , Pueraria , Genetics
12.
Article in Chinese | WPRIM | ID: wpr-773673

ABSTRACT

Armillaria gallica is a symbiotic fungus in the cultivation process of Gastrodia elata and Polyporus.The rhizomorph of A. gallica invades the stalk of the G. elata or the Sclerotium of the Polyporus,and is digested and utilized by the latter,becoming their important source of nutrition. Different nature of A. gallica affects the growth of G. elata and Polyporus. The authors collected A. gallica from 13 commercially available regions and screened two A. gallica,A and B,at the genetic and metabolic levels,in order to distinguish between the two A. gallica market. We have established convenient and effective DNA molecular identification method.By comparing the sequence differences between the A. gallica type A and type B invertase genes,PCR-RFLP primers were designed based on differential fragment. Primer ZTM.F/ZTM.R can amplified A. gallica type A and B,producing a band of about 304 bp in length. The restriction endonuclease EcoR V could recognize the difference sequence of A and B types of A. gallica. The type B was digested to form two fragments,thereby specifically identifying the A. gallica as type B. The established methods of PCR-RFLP is an accurate identification method for A. gallica. Therefore,in the cultivation process of G. elata and Polyporus,suitable strains can be selected according to different needs of variety,growth stage and ecological environment,and the yield and quality can be improved according to local conditions.


Subject(s)
Armillaria , Classification , Gastrodia , Microbiology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Polyporus
13.
Article in Chinese | WPRIM | ID: wpr-690707

ABSTRACT

A total of 58 varieties in Lonicera japonica from 20 producing areas were amplified by 22 pairs of SSR primers. Seven pairs of polymorphic primers were screened and their primers were used to establish DNA identity card and analyze genetic similarity.All the 58 varieties could be distinguished each other by the DNA identity card constituted by 7 pairs of core SSR primers.The genetic similarity coefficients of 58 varieties ranged from 0.366 7 to 0.916 7 by using PopGene32(vesion1.32). Furthermore, all the varieties consistency were classified into 4 groups and constructed an evaluation table according to cluster analysis by an un-weighted pair-group average method with arithmetic mean. As expected, the results of cluster and evaluation table reflected 58 varieties relatives, which provide reference information for the selection of fine germplasm of L. japonica and the theoretical basis for the study of Dao-di herbs.

14.
Article in Chinese | WPRIM | ID: wpr-775319

ABSTRACT

This study is to establish a pre-column derivatization procedure with 1-phenyl-3-methyl-5-pyrazolone (PMP) UPLC-MS/MS method for the determination of the monosaccharide composition of 12 polysaccharides. At the same time, the monosaccharide components of polysaccharides in Armillaria gallica were analyzed. The separation was performed on a ACQUITY ZORBAX RRHD Eclipse Plus C₁₈ column(2.1 mm×100 mm, 1.8 μm),using 95% acetonitrile (A) and ammonium acetate-5% acetonitrile-water (B) as mobile phase with gradient elution. The target components were detected in multiple-reaction monitoring (MRM) mode by mass spectrometry with electrospray ionization (ESI) source operated in ionization mode. The results showed that based on the monosaccharides detection method established by UPLC-MS/MS, the linearity of the 12 monosaccharides components were linear in their linear range (R²>0.990), and the recovery rate were 92.30%-105.6%. 11 monosaccharides such as fructose, mannose, and glucose were detected in A. gallica samples. The method established in this experiment is robust, highly reproducible and accurate, and is suitable for the determination of monosaccharide components such as A. gallica.


Subject(s)
Chromatography, High Pressure Liquid , Chromatography, Liquid , Monosaccharides , Polysaccharides , Tandem Mass Spectrometry
15.
Acta Pharmaceutica Sinica ; (12): 1202-1208, 2018.
Article in Chinese | WPRIM | ID: wpr-779989

ABSTRACT

Plant flowering regulation is an important mechanism to response to environmental stress. Heat shock protein 70 family is one of the main molecular chaperones to resist stress; miRNA can be used as a negative regulator to participate in post-transcriptional gene in flowering network. In this paper, we obtained an Hsp70 gene from Lonicera japonica transcriptome and combined with Lonicera japonica miRNA library to obtain a novel miRNA that may target Hsp70 gene through bioinformatics method. Bioinformatics and expression during different flowering stages of the obtained Hsp70 gene and miRNA were analyzed. Phylogenetic tree showed that the obtained Hsp70 gene was clustered with Hsp110 subfamily in Oryza sativa and Arabidopis thaliana. The prediction of miRNA secondary structure showed its stable structure and high reliability. The binding site map showed that there were two base mismatches between sequences of miRNA and Hsp70 gene. The expression analysis showed that the expression of Hsp70 and miRNA in different flowering stages had opposite trends, indicating that miRNA might regulate Hsp70 to participate in the flowering stages of Lonicera japonica. This study provided new ideas for Lonicera japonica flowering regulation and response to environmental stress mechanisms.

16.
Acta Pharmaceutica Sinica ; (12): 1770-1776, 2017.
Article in Chinese | WPRIM | ID: wpr-779788

ABSTRACT

Auxin response factor (ARF) is an important transcription factor for auxin signal transduction pathway, which regulates virtually every aspect of plant growth and development from embryogenesis to senescence. Nine full-length genes of ARF transcription factors were obtained from transcriptome dataset of Scutellaria baicalensis Georgi using the bioinformatics methods. The nucleotide and protein characteristics, subcellular localization, senior structural domains and conservative forecasts of those ARF transcription factors were analyzed. The phylogenetic tree showed that the nine ARFs in S. baicalensis were clustered together with ARF transcription factors in Arabidopsis thaliana, Oryza sativa subsp. Japonica and Nicotiana attenuate. The results of gene expression showed that:① The expression levels of ARF1, ARF3, ARF4, ARF8, ARF20 and ARF24 were upregulated after 100 μmol·L-1 GA3 treatment. However, the expression levels of ARF6 and ARF18 were downregulated; ② Those ARF genes were mainly expressed in the flowers of S. baicalensis; ③ There was a significant correlation between ARF genes and the genes involved in flavonoid biosynthesis. Our results provide a basis for further understanding the molecular regulation mechanism of flavonoid biosynthesis in S. baicalensis.

17.
Article in Chinese | WPRIM | ID: wpr-256015

ABSTRACT

Medicinal Polyporus umbellatus is the dry sclerotia of P. umbellatus, with the effect of diuresis; Armillaria mellea is a parasitic fungus which can infect plants up to 300 genera, with sedative, anticonvulsant and some other biological activities. As the medicinal value of P. umbellatus and A. mellea is increasingly wide concerned, the market quantity demanded of them is gradually increased and the demand outstrips the supply. The symbiotic A. mellea and P. umbellatus are both the medicinal and edible fungi with diverse activities, including hypoglycemic action, improve immunity and antitumor and so on. The growth of the sclerotia forming from the mycelium of P. umbellatus is related to the infection of the symbiotic A. mellea and their secondary products. In this study, by comparing the chemical constituents of the mycelium and sclerotia of P. umbellatus and A. mellea, we found that they all produced steroids and nitrogen-containing heterocycles. The sclerotia of P. umbellatus and A. mellea also produced triterpenes secondary metabolites. In addition, the mycelium and infected sclerotia of P. umbellatus mainly produced different steroids, and the sclerotia produced some other special secondary metabolites, such as long-chain fatty acids, ceramides, phenol and so on. By analyzing above all kinds of differences, speculated that these may be caused by the infection of the symbiotic A. mellea which mainly produced sesquiterpenes, diterpenes and other secondary metabolites. The contents and types of compounds of P. umbellatus and A. mellea are closely related to their symbiosis and reproduction, therefore, many symbiosis mechanisms should be found by utilizing more molecular biology technology to elucidate this complex symbiotic infection and provide scientific basis for improving the yield and quality of P. umbellatus and A. mellea.

18.
Article in Chinese | WPRIM | ID: wpr-304880

ABSTRACT

The stress effect is a characteristic of Dao-di herbs caused by environmental gene expression of medical plants is influenced by environmental changes and finally affects the formation and accumulation of metabolites. Using Scutellaria baicalensis as material, active component of wild type of S. baicalensis from 19 production areas were analysed; It was found that climate change can influence the accumulation of active components. Then, S. baicalensis suspension cells was exposed to various environments, and enzyme activity and gene expression were measured, indicating the molecular mechanism of stress effect on S. baicalensis. Hence, we found the prerequisite and method to study the stress effect on Dao-di herbs, and we hope this research can provides some references for another studies of Dao-di herbs.

19.
Article in Chinese | WPRIM | ID: wpr-231018

ABSTRACT

To establish a new high resolution melting analytical method for identification of Lonicera japonica germplasm, the screening of 7 pairs of SSR (simple sequence repeats) primers, determining the suitable diagnostic primers by the differences of peak pattern and Tm was conducted. Then into the DNA template concentration, annealing temperature and the suitable range of cycle number were investigated. Combined with SIMCA-P software for data processing analysis, the results show that three main germplasm honeysuckle could be divided by four sets of primers. It provides methodology for improving L. japonica germplasm identification.

20.
Article in Chinese | WPRIM | ID: wpr-294336

ABSTRACT

<p><b>OBJECTIVE</b>To explore the effect of curcumin (CUR) on cycteinyl aspirate specific protease-12 (Caspase-12) and pneumocyte apoptosis in pulmonary ischemia/reperfusion (I/R) injury mice.</p><p><b>METHODS</b>The in vivo unilateral in situ pulmonary I/R injury mouse model was established in C57BL/6J mice. Sixty experimental mice were randomly divided into six groups by random digit table, i. e., the sham-operation group (Sham), the I/R group, the I/R + dimethyl sulfoxide group (I/R + DMSO), the I/R + low dose CUR pre-treated group (I/R + CUR-100), the I/R + middle dose CUR pre-treated group (I/R + CUR-150), the I/R + high dose CUR pre-treated group (I/R + CUR-200), 10 in each group. Mice were euthanized and their left lungs were excised. Wet lung weight to dry lung weight (W/D) and the total lung water content (TLW) were tested. The morphological changes of the lung tissue were observed and index of quantitative evaluation for alveolar damage (IQA) detected under light microscope. The ultra-microstructure of the lung tissue was observed under electron microscope. The mRNA and protein expression levels of Caspase-12 and glucose regulated protein (GRP78) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. Apoptosis index (AI) of the lung tissue was determined by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) method.</p><p><b>RESULTS</b>Compared with the Sham group, expression levels of Caspase-12, GRP78 mRNA and protein all significantly increased in the I/R group (P < 0.05); W/D, TLW, IQA, and AI were all notably higher (P < 0.05, P < 0.01); the morphological and ultrastructural injury of the lung tissue were notably observed in I/R group. Compared with the I/R + DMSO group, expression levels of GRP78 mRNA and protein were increasingly higher in the I/R + CUR-100 group, the I/R + CUR-150 group, and the I/R +CUR-200 group (P < 0.05), expression levels of Caspase-12 mRNA and protein were lower (P < 0.05); W/D, TLW, IQA, and AI also decreased (P < 0.05, P < 0.01); the morphological and ultrastructural injury of the lung tissue were gradually alleviated in the I/R + CUR groups.</p><p><b>CONCLUSION</b>CUR had better effect on the lung protection against I/R injury, which might be related to inhibition for pneumocyte apoptosis associated with Caspase-12 in excessive unfolded protein response (UPR).</p>


Subject(s)
Animals , Apoptosis , Caspase 12 , Metabolism , Curcumin , Pharmacology , Heat-Shock Proteins , Metabolism , Lung , Male , Mice , Mice, Inbred C57BL , Reperfusion Injury , Metabolism , Pathology
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