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1.
Acta Pharmaceutica Sinica ; (12): 525-532, 2022.
Article in Chinese | WPRIM | ID: wpr-922932

ABSTRACT

In this study, nrDNA ITS sequences of Lycium cultivars were sequenced and used to test the existence of incomplete concerted evolution and pseudogenes. Together with 44 ITS sequences retrieved from GenBank, the pattern of base substitutions, GC content, 5.8S conserved motifs, the minimum free energy of secondary structures, nucleotide diversity and phylogenetic relationship of the samples were analyzed. While 83 of the 144 sequences were identified as pseudogenes, the results suggested a high degree of polymorphism and putative pseudogenes in Lycium, suggesting an incomplete concerted evolution of the ITS region. ITS polymorphism and pseudogene of Lycium were systematically tested for the first time. This research provides a references for ITS sequence to be used in the study of Lycium germplasm resources and DNA barcode identification.

2.
Article in Chinese | WPRIM | ID: wpr-909592

ABSTRACT

OBJECTIVE Programmed death ligand-1 (PD-L1) and indoleamine 2, 3-dioxygenase 1 (IDO1) are immune checkpoints which can be induced by interferon-γ(IFN-γ) in the tumor microenvironment, leading to immune escape of tumors. Myricetin (MY) is a flavonoid distributed in many edible and medicinal plants. The aim of this study is to clarify the effect and the mechanism of MY on inhibiting IFN-γ-induced PD-L1 and IDO1 in lung cancer cells. METHODS Expressions of PD-L1 and major histocompatibility complex-I (MHC-I) were evaluated by flow cytometry and Western blotting, and the expression of IDO1 was measured by Western blotting. qRT-PCR was used to detect their mRNA levels. The function of T cells was evaluated using a co-culture system consist of lung cancer cells and the Jurkat-PD-1 T cell line that overexpressing PD-1. Molecular docking analysis, Western blotting and immunofluorescence were used for mechanism study. RESULTS MY potently inhibited IFN-γ-induced PD-L1 and IDO1 expression in human lung cancer cells, while didn't show obvious effect on the expression of MHC-I. In addition, MY restored the survival, proliferation, CD69 expression and interleukin-2 (IL-2) secretion of Jurkat-PD-1 T cells suppressed by IFN-γ-treated lung cancer cells in the co-culture system. Mechanistically, IFN-γ up-regulated PD-L1 and IDO1 at the transcriptional level through the JAK-STAT-IRF1 axis, which was targeted and inhibited by MY. CONCLUSION Our research revealed a new insight into the anti-tumor effects of MY which inhibited IFN-γ-induced PD-L1 and IDO1 expression, supporting the potential of MY in anti-tumor immunotherapy.

3.
Acta Pharmaceutica Sinica ; (12): 3285-3299, 2021.
Article in Chinese | WPRIM | ID: wpr-906851

ABSTRACT

Plant natural products (PNPs) are important sources of innovative drugs. They are mainly obtained by isolation or extraction from plants. Low content and with structural analogues in plants result in high production cost, which restricts the research and application of PNPs. While biopathway construction by synthetic biology provides an alternative for production of PNPs. By biosynthetic pathway analysis of PNPs and reconstructing the biopathway in microorganisms, we can produce PNPs in cell factories efficiently. Recently, several predominantly international reports about biosynthesis of PNPs and its synthetic biology production, triggered the researches of PNPs. Abundant traditional Chinese medicine resources and profound cultural heritage of Chinese medicine make biosynthesis pathway analysis of PNPs to be a research hotspot. And some of the studies have achieved significant progress. Here, recent progress in the biosynthesis of plant natural products and its synthetic biology was reviewed. In particular, the application of new methods and technologies in recent years were summarized and discussed. This will provide reference for the biopathway construction of plant natural products.

4.
Article in Chinese | WPRIM | ID: wpr-906444

ABSTRACT

The historical evolution, fermentation technology and key links of Sojae Semen Praeparatum (SSP) were sorted out by consulting ancient books and modern literature, and the influencing factors and control methods of quality were analyzed and summarized in order to provide reference for the quality control of SSP. After analysis, it was found that in the fermentation process of SSP, fermentation strains, miscellaneous bacteria, temperature and humidity were all important factors affecting the quality of SSP. The condition control of "post fermentation" process has been paid more attention to in the past dynasties. In addition, the delicious SSP recognized in ancient times should be made from mold fermentation, and the breeding and application of fermented mold may be the key point to solve the quality problem of SSP. Therefore, based on the evaluation indexes of SSP in the past dynasties, it is of great significance to study and optimize the technological conditions such as strain, temperature and humidity in depth to improve the quality of SSP.

5.
Article in Chinese | WPRIM | ID: wpr-906401

ABSTRACT

Objective:To compare the contents of adenosine, gastrodin, <italic>p</italic>-hydroxybenzyl alcohol, <italic>p</italic>-hydroxybenzaldehyde, parisinin B and parisinin A in Chijian (the aerial part of <italic>Gastrodia elata</italic>) and Gastrodiae Rhizoma, and compare their effects on immune function and intestinal microflora, evaluating whether it is necessary to study and develop Chijian. Method:The contents of these six constituents were determined by ultra performance liquid chromatography (UPLC), the mobile phase was 0.1% formic acid aqueous solution (A)-acetonitrile (B) for gradient elution (0-4 min, 0.5%B; 4-5 min, 0.5%-2%B; 5-10 min, 2%-15%B; 10-12 min, 15%-20%B; 12-15 min, 20%-95%B; 15-17 min, 95%B; 17-17.5 min, 95%-0.5%B; 17.5-20 min, 0.5%B), the flow rate was 0.5 mL·min<sup>-1</sup>, the detection wavelength was 270 nm. The difference of pharmacological activity of water extracts of Chijian and Gastrodiae Rhizoma was compared, the clearance index, corrected clearance index and peripheral blood were measured in mice model with low immune function induced by cyclophosphamide, B lymphocyte proliferation was determined by lymphocyte transformation test <italic>in vitro</italic>, intestinal microflora was analyzed by 16S rDNA technology and bioinformatics was conducted. Result:The total contents of these six components in powder and ethanol extract of Chijian were higher than that of Gastrodiae Rhizoma, but the total contents of these six components in their water extract were similar, and the total contents of gastrodin and <italic>p</italic>-hydroxybenzyl alcohol met the requirements of the 2020 edition of <italic>Chinese Pharmacopoeia</italic>. Compared with the blank group, the clearance index of immunocompromised mice was significantly increased in the middle-dose (10 g·kg<sup>-1</sup>) group of Chijian water extract, middle- and low-dose (10, 5 g·kg<sup>-1</sup>) groups of Gastrodiae Rhizoma water extract (<italic>P</italic><0.05), the levels of erythrocyte and hematocrit in peripheral blood were significantly increased in the high-dose (20 g·kg<sup>-1</sup>) groups of water extracts of Chijian and Gastrodiae Rhizoma (<italic>P</italic><0.05, <italic>P</italic><0.01), water extract of Gastrodiae Rhizoma with concentration of 400 g·L<sup>-1</sup> and the water extract of Chijian with the concentration of 100 g·L<sup>-1</sup> could promote the proliferation of B lymphocytes induced by lipopolysaccharide. Studies on intestinal microflora showed that compared with the blank group, at the phylum level, the water extracts of Chijian and Gastrodiae Rhizoma increased the relative abundance of Bacteroidetes and decreased the relative abundance of Firmicutes, at the genus level, they increased the relative abundance of <italic>Prevotellaceae</italic>_UCG-001 and <italic>Ruminococcaceae</italic>_UCG-005, and decreased the relative abundance of <italic>Anaerotruncus</italic>, unclassified_<italic>f</italic>_<italic>Erysipelotrichaceae</italic> and<italic> Candidatus</italic>_<italic>Stoquefichus</italic>.<italic> </italic>These intestinal bacteria were related to the immune system, cell proliferation, and metabolism regulation. Conclusion:The total contents of 6 components in the powder, the ethanol and the water extracts of Chijian are higher than or close to those of the corresponding samples of Gastrodiae Rhizoma, the pharmacological activity of Chijian water extract is similar to that of Gastrodiae Rhizoma water extract, indicating that Chijian is worthy of further research and development.

6.
Article in Chinese | WPRIM | ID: wpr-906365

ABSTRACT

Objective:The chemical constituents in guarana (<italic>Paullinia cupana</italic> dried seeds) were systematically analyzed to provide a basis for further research, development and utilization of this plant. Method:The contents of crude protein, crude fat, crude polysaccharide and crude fiber in guarana were determined according to national standards and related documents, and the chemical constituents of guarana was qualitatively analyzed by ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS), ACQUITY UPLC-HSS-T3 column (2.1 mm×100 mm, 1.8 μm) was used with 0.1% formic acid aqueous solution (A)-0.1% formic acid acetonitrile solution (B) as mobile phase for gradient elution (0-5 min, 2%-10%B; 5-6 min, 10%-20%B; 6-9 min, 20%-30%B; 9-9.5 min, 30%-35%B; 9.5-10.5 min, 35%-45%B; 10.5~13 min, 45%-55%B; 13-15 min, 55%-80%B; 15-19 min, 80%-98%B; 19-20 min, 98%B; 20-20.3 min, 98%-2%B; 20.3-23 min, 2%B), the electrospray ionization (ESI) was used for detection in positive and negative ion modes, the scanning range was <italic>m</italic>/<italic>z</italic> 50-1 500, and the structure was identified according to the relative molecular weight and fragment information combined with database matching and comparison of reference substances. Result:The contents crude protein, crude fat, crude polysaccharide and crude fiber in guarana were (0.63±0.03)%, (2.73±0.09)%, (3.23±0.12)% and (8.89±0.59)%, respectively. A total of 42 chemical constituents in guarana were identified by UPLC-Q-TOF-MS, including 3 methylxanthines, 2 nucleosides, 1 amino acid, 3 organic acids, 33 flavonoids, 3 (<italic>L</italic>-tryptophan, epigallocatechin gallate, daidzein) of which were first discovered in guarana. Conclusion:Guarana is rich in nutrients and has good potential to be developed as a functional food. UPLC-Q-TOF-MS technique provides a simple, rapid and accurate method for the identification of chemical constituents in guarana. Methylxanthines and proanthocyanidins are the main chemical constituents of guarana, which is meaningful for quality evaluation and material basis of guarana.

7.
Article in Chinese | WPRIM | ID: wpr-905914

ABSTRACT

Literature research on ancient books of traditional Chinese medicine (TCM) is an important carrier for inheriting the academic achievements and thoughts of TCM, and a key step for continuing the Chinese civilization and realizing the great rejuvenation of the Chinese nation. Based on this, the paper puts forward the purpose of sorting out TCM ancient books:to explore the treasure of traditional culture, reveal its significance, carry forward its spirit, learn from its experiences, so as to make a contribution to the development of TCM. And this paper expounds several major problems in the literature research, that is, paying attention to the phenomenon of "stubborn bass", avoiding the trend of "latecomers turning inferior", attaching importance to the hidden trouble of "making comments on behalf of the ancients". Then, this paper discusses the methodology of carrying out accurate research and revealing the true nature and true value of scholarship with the idea of confucian orthodoxy, the rules of not forgetting the original intention and the scientific method. Taking the materia medica archaeology as an example, the author shared the practical exploration of how to crack the historical code with scientific and technological means, so as to provide useful reference for the literature research on TCM ancient books.

8.
Article in Chinese | WPRIM | ID: wpr-921788

ABSTRACT

Lancang-Mekong Cooperation is a new type of subregional cooperation mechanism initiated and built by China and other five countries of the Lancang-Mekong subregion, namely Laos, Myanmar, Thailand, Cambodia, and Vietnam. Countries in the Lancang-Mekong subregion are geographically and culturally connected, and they have nurtured their unique traditional medicine. By combing the history of traditional medicine exchanges between China and other Lancang-Mekong countries and their progress of modern research, this paper summarized the challenges and opportunities of traditional medicine cooperation in the Lancang-Mekong subregion. It has been found that many regional cooperation mechanisms coexist for a long time in the Lancang-Mekong subregion and the medicinal resources are abundant. However, the degree of their development and utilization varies, and modern scientific research is insufficient. Lancang-Mekong Cooperation has provided a strong support for integrating the advantageous resources in Lancang-Mekong subregion countries and making progress together. Focusing on the development and protection of medicinal resources, this paper puts forward a new path of cooperation in the intellectual property rights and characteristic seed resource protection, the compilation of universal herbal pharmacopoeia in various countries, the research and development of public health products, and the construction of traditional herbal industry bases, thus enabling the traditional medicine to better protect the public health and building a human health community.


Subject(s)
China , Humans , Materia Medica , Medicine, Traditional , Rivers , Thailand
9.
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
10.
Article in English | WPRIM | ID: wpr-921356

ABSTRACT

Objective@#Early triage of patients with coronavirus disease 2019 (COVID-19) is pivotal in managing the disease. However, studies on the clinical risk score system of the risk factors for the development of severe disease are limited. Hence, we conducted a clinical risk score system for severe illness, which might optimize appropriate treatment strategies.@*Methods@#We conducted a retrospective, single-center study at the JinYinTan Hospital from January 24, 2020 to March 31, 2020. We evaluated the demographic, clinical, and laboratory data and performed a 10-fold cross-validation to split the data into a training set and validation set. We then screened the prognostic factors for severe illness using the least absolute shrinkage and selection operator (LASSO) and logistic regression, and finally conducted a risk score to estimate the probability of severe illness in the training set. Data from the validation set were used to validate the score.@*Results@#A total of 295 patients were included. From 49 potential risk factors, 3 variables were measured as the risk score: neutrophil to lymphocyte ratio ( @*Conclusion@#This report may help define the potential of developing severe illness in patients with COVID-19 at an early stage, which might be related to the neutrophil to lymphocyte ratio, albumin, and chest computed tomography abnormalities.


Subject(s)
Aged , COVID-19/diagnosis , Female , Humans , Male , Middle Aged , Nomograms , Retrospective Studies , Risk Assessment , Severity of Illness Index
11.
Article in Chinese | WPRIM | ID: wpr-888136

ABSTRACT

Safflower(Carthamus tinctorius), a valuable traditional Chinese medicinal plant, has attracted much attention in recent years. This study established a stable tissue culture system of safflower and analyzed the chromatogram of its secondary metabolites, providing high-quality experimental materials for further research on natural products in safflower. The calluses were established from the safflower seeds germinated in a sterile environment, and then they were differentiated into the aseptic seedlings, or cultured to obtain suspension cells in liquid medium. The ultra-performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF-MS), Progenesis QI, and principal component analysis(PCA) were used to detect and analyze the secondary metabolites in the suspension cells before and after induction with different elicitors(methyl jasmonate, silver nitrate, salicylic acid and yeast extract). A total of 23 secondary metabolites including flavonoids, phenylpropanoids, alkaloids, fatty acids and aromatic glycosides were detected in safflower suspension cells. In response to the four elicitors, 11 compounds showed increased or decreased relative content. The results indicate that different elicitors have various effects on the accumulation of secondary metabolites in safflower suspension cells, and yeast extract shows more obvious positive induction. Therefore, different elicitors may play a role in the expression of related genes in the biosynthetic pathway of specific secondary metabolites. The results facilitate the discovery of targeted elicitors and the large-scale production of valuable secondary metabolites in the future.


Subject(s)
Carthamus tinctorius , Chromatography, High Pressure Liquid , Chromatography, Liquid , Flavonoids , Glycosides , Mass Spectrometry
12.
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
13.
Article in Chinese | WPRIM | ID: wpr-888103

ABSTRACT

Volatile oil is the main effective component and an important quality indicator of Artemisia argyi leaves. In this study, 100 germplasm resources of A. argyi were collected from all the related habitats in China. The total volatile oils in A. argyi leaves were extracted by steam distillation and the content was determined by GC-MS. The result demonstrated that the content of total volatile oils was in the range of 0.53%-2.55%, with the average of 1.43%. A total of 39 chemical constituents were identified from the volatile oils, including 13 shared by the 100 germplasm resources. Clustering analysis of the 39 constituents showed that the 100 A. argyi samples were categorized into groups Ⅰ(9), Ⅱ(2), Ⅲ(66) and Ⅳ(23), and group Ⅲ had the most volatile medicinal components, with the highest content. Five principal components(PCs) were extracted from 13 shared constituents, which explained 73.454% of the total variance. PC1, PC2, and PC3 mainly reflected the pharmacological activity of volatile oils and the rest two the aroma information. The volatile oils identified in this study lay a foundation for variety breeding of and rational utilization of volatile oils in A. argyi leaves.


Subject(s)
Artemisia , Distillation , Oils, Volatile , Plant Breeding , Plant Leaves
14.
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
15.
Article in Chinese | WPRIM | ID: wpr-887996

ABSTRACT

Chinese materia medica( CMM) serves as an important cornerstone for the development of traditional Chinese medicine( TCM) culture and industry due to its unique ecological,medical,economic,scientific and technological,and cultural values. The supply shortage and unstable quality of some CMM resources have hindered the development of TCM. Ensuring the sustainable use of CMM resources has become essential for the development of TCM in China. Enriching CMM resources is the key to ensuring the sustainable utilization of TCM resources in China,which can be achieved via expanding the medicinal parts,developing the substitutes,seeking for analogues,exploring the ethnic and folk medicines,or introducing foreign medicinal materials. CMM efficacy or function positioning plays a very important role in the transformation of new CMM resources. The strategies and methods for efficacy or function positioning of new CMM resources,including analogy,plant genetic relationship exploration,medicinal property deduction,ethnobotanical investigation,text mining,network pharmacology,and structure-activity relationship exploration,were systematically proposed in this study based on CMM theory,textual research,and modern methodologies. This paper is expected to provide a theoretical reference for the continuous enrichment and development of CMM resources and the high-quality development of TCM culture and industry.


Subject(s)
China , Data Mining , Drugs, Chinese Herbal/pharmacology , Humans , Materia Medica , Medicine, Chinese Traditional
16.
Article in Chinese | WPRIM | ID: wpr-887981

ABSTRACT

The basic features of glandular and non-glandular trichomes on leaves of Artemisia argyi( germplasms from Qichun,Ningbo,Tangyin,and Anguo,respectively) and related species A. stolonifera were observed by scanning electron microscopy( SEM)and compared. There were significant differences in trichome characteristics of leaves at all parts of A. argyi and A. stolonifera,which were closely related to the difference in chemical components. The length of non-glandular trichomes and size of glandular trichomes on middle leaves were the stablest. A. argyi and A. stolonifera can be distinguished by the density of glandular trichome. Additionally,the four germplasms of A. argyi can be discriminated via the density and curvature of non-glandular trichome. The density of non-glandular trichomes was the highest in A. stolonifera. For A. argyi,the germplasm from Qichun had the highest density of non-glandular trichomes on the abaxial surfaces of upper leaves and that from Ningbo had the largest non-glandular trichome curvature. With regard to the germplasm from Anguo,the T-shaped non-glandular trichomes of long stalks on the adaxial surfaces of the middle leaves were lodging-susceptible,and those with slender heads were wave-like. Statistics results of A. argyi and A. stolonifera are as follows: largest glandular trichomes on the adaxial and abaxial surfaces and highest glandular trichome density on the abaxial surfaces of the lower leaves in A. argyi germplasm from Ningbo,highest density of non-glandular trichomes on the abaxial surfaces of upper leaves in A. stolonifera,and highest density of glandular trichomes and non-glandular trichomes on the adaxial surfaces of the upper leaves in A. argyi germplasm from Qichun. According to the observation result under fluorescence microscope( FM),flavonoids were closely related to the size and density of non-glandular trichomes and size of glandular trichomes. The fluorescence intensity was the strongest and fluorescence area was the largest for flavonoids in A. argyi germplasms from Qichun and Tangyin,while the fluorescence for flavonoids was the weakest in A. stolonifera. It was the first time to observe and analyze the trichome ultrastructure of A. argyi leaves at different positions by SEM and FM. This study clarifies the differences between A. stolonifera and four famous A. argyi germplasms,which provides new evidence for the microscopic identification of A. argyi and its related species and serves as a reference for the study of the relationship of A. argyi structure with its components and functions.


Subject(s)
Artemisia , Flavonoids , Microscopy, Electron, Scanning , Plant Leaves , Trichomes
17.
Article in Chinese | WPRIM | ID: wpr-887953

ABSTRACT

The plant root-associated microbiomes include root microbiome and rhizosphere microbiome, which are closely related to plant life activities. Nearly 30% of photosynthesis products of plants are used to synthesize root compounds, there is evidence that root compounds regulate and significantly affect the root microbiome Tanshinones are the main hydrophobic components in Salvia miltiorrhiza. In order to study whether these compounds can regulate the root-associated microbiomes of S. miltiorrhiza, our study first identified a white root S. miltiorrhiza(BG) which contains little tanshinones. Retain of the fifth intron of tanshinones synthesis key enzyme gene SmCPS1 leading to the early termination of the SmCPS1 gene, and a stable white root phenotype. Further, wild type(WT) and BG were planted in greenhouse with nutrient soil(Pindstrup, Denmark) and Shandong soil(collected from the S. miltiorrhiza base in Weifang, Shandong), then high-throughput sequencing was used to analyze the root-associated microbiomes. The results showed that the tanshinones significantly affected the root-associated microbiomes of S. miltiorrhiza, and the impact on root microbiomes was more significant. There are significant differences between WT and BG root microbiomes in species richness, dominant strains and co-occurrence network. Tanshinones have a certain repelling effect on Bacilli which belongs to Gram-positive, while specifically attract some Gram-negative bacteria such as Betaproteobacteria and some specific genus of Alphaproteobacteria. This study determined the important role of tanshinones in regulating the structure of root-associated microbiomes from multiple angles, and shed a light for further improving the quality and yield of S. miltiorrhiza through microenvironment regulation.


Subject(s)
Abietanes , Microbiota , Plant Roots , Salvia miltiorrhiza
18.
Article in Chinese | WPRIM | ID: wpr-887944

ABSTRACT

Drynariae Rhizoma is warm in nature and bitter in taste, mainly acting on liver and kidney systems. It is a common Chinese herbal medicine for the treatment of fracture and bone injury. The chemical compositions of Drynariae Rhizoma mainly include flavonoids, triterpenoids, phenylpropanoids and lignans. At present, modern pharmacological and clinical studies have shown that Drynariae Rhizoma has the effects of anti osteoporosis, promoting fracture healing, kidney protection, anti-inflammatory, promoting tooth growth, preventing and treating aminoglycoside ototoxicity and lowering blood lipid. In addition, the toxicity evaluation experiment of Drynariae Rhizoma has also shown that it has no obvious toxic and side effects. Naringin is a kind of dihydroflavone in Drynariae Rhizoma. Many studies have shown that naringin and other total flavonoids play an important role in anti-osteoporosis, promoting fracture healing, anti-inflammation, promoting tooth growth and lowering blood lipid. In this study, the research progresses on chemical consti-tuents and pharmacological activities of Drynariae Rhizoma in recent years were reviewed, and some mechanisms of action were summarized, to provide references for the further research and development of Drynariae Rhizoma.


Subject(s)
Drugs, Chinese Herbal/pharmacology , Flavonoids , Humans , Osteoporosis/drug therapy , Polypodiaceae , Rhizome
19.
Acta Pharmaceutica Sinica ; (12): 1478-1485, 2021.
Article in Chinese | WPRIM | ID: wpr-887073

ABSTRACT

In this study, the fatty acid desaturase gene FAD2 was cloned from Coix lacryma-jobi L. and its molecular structure and function were studied. The results showed that the full-length cDNA sequence of FAD2 gene was 936 bp encoding 311 amino acid residues. Bioinformatics prediction results showed that the protein encoded by the FAD2 gene was an alkaline hydrophilic unstable protein with a molecular weight of 34.87 kDa. It contained three transmembrane helix domain, and did not contain the signal peptide splicing site, and was most likely to be located in plasmid membrane. Compared with other similar genes in plants, it has only a histidine conserved site, His Box Ⅲ histidine site (HXXHH), suggesting its activity may be reduced. Phylogenetic tree analysis showed that FAD2 was closely related to monocotyledonous plants, especially Maize and Oryza sativa japonica Group, but farther from dicotyledonous plants. Therefore, it was inferred that FAD2 might have similar functions with similar genes in Maize and Oryza sativa japonica Group. In addition, the expression of FAD2 gene could be detected in Coix lacryma-jobi L. with high oil content, but not in low oil content of Coix lacryma-jobi L. In order to clarify the function of FAD2, the gene was heterologously expressed in sporomyces cerevisiae. The results showed that the protein encoded by FAD2 gene did not catalyze the formation of C18∶1 unsaturated fatty acid into C18∶2 unsaturated fatty acid. Therefore, it was speculated that the deletion of histidinine conserved site of FAD2 gene might lead to the decrease of protein activity or even inactivation. This study provides reference value for further understanding the molecular structure characteristics of fatty acid desaturase. At the same time, it laid a foundation for elucidating the biosynthetic pathway of Coix lacryma-jobi L.

20.
Article in Chinese | WPRIM | ID: wpr-879176

ABSTRACT

Carboxyl CoA ligases(CCLs) is an important branch of adenylate synthetase gene family, which mainly has two-step catalytic reactions. Firstly, in the presence of adenosine triphosphate, it can catalyze the pyrophosphorylation of carboxylateswith diffe-rent structures to form corresponding acyl adenosine monophosphate intermediates. Secondly, adenosine monophosphate was replaced by free electrons in the mercaptan group of enzyme A or other acyl receptors by nucleophilic attack to form thioesters. In this study, on the basis of the transcriptome database of Arnebia euchroma, two genes were selected, named AeCCL5(XP_019237476.1) and AeCCL7(XP_019237476.1). Bioinformatics analysis showed that their relative molecular weights were 60.569 kDa and 60.928 kDa, theoretical PI were 8.59 and 8.92, respectively. They both have transmembrane domains but without signal peptide. By multiple sequence alignment and phylogenetic tree analysis, we found that the similarity between AeCCLs and other plant homologous proteins was not high, and the substrate binding sites of AeCCLs were not highly conserved. The reasons might be that the sequence and structure need to adapt to the changes of new substrates in the process of evolution. In this study, the full-length of AeCCL5 and AecCCL7 were cloned into the expression vector pCDFDuet-1. The proteins of AeCCL5 and AeCCL7 with His-tag were expressed in Escherichia coli. The proteins of AeCCL5 and AeCCL7 were purified by nickel column. In vitro enzymatic reactions proved that both AeCCL5 and AeCCL7 can participate in the upstream phenylpropane pathway of shikonin biosynthesisby catalyzing 4-coumaric acid to produce 4-coumarin-CoA, and then to synthesis p-hydroxybenzoic acid, which is an important precursor of shikonin biosynthesis in A. euchroma.


Subject(s)
Boraginaceae/genetics , Cloning, Molecular , Coenzyme A , Coenzyme A Ligases/genetics , Ligases , Phylogeny
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