1.Pueraria lobate advances in molecular pharmacognosy.
Bi-Sui YANG ; Qiu-Lian HUANG ; Lu-Xin XIE ; Bo WU ; Ke-Zhong DENG ; Zhi-Gui WU ; Wei-Feng ZHU ; Shao-Lang HE ; Qi HUANG ; Yu-Ye ZHU ; Fei GE
China Journal of Chinese Materia Medica 2021;46(9):2149-2157
Molecular pharmacognosy is a science of classification and identification, cultivation and protection, and production of active ingredients of graduated drugs at the molecular level. The proposal of molecular pharmacognosy allows the research of crude drugs to advance from the microscopic level to the genetic level. Pueraria lobata root, as a medicinal and edible plant, has high application value and economic value. There are many varieties that are easy to cause confusion, and it is not easy to distinguish and identify according to traditional identification methods. Moreover, the research of P. lobate root at the genetic level is still relatively shallow. the study received extensive attention of scholars. This article reviews recent research on molecular identification of P. lobate, transcriptome sequencing, cloning and synthesis of functional genes of P. lobate root in recent years in order to provide references for further promoting the development and utilization of P. lobate root and its active ingredients.
Pharmacognosy
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Plant Roots/genetics*
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Pueraria
2.Karyotypes analysis of Ferula fukanensis.
Xin ZHAO ; Xiao-Jun MA ; Sulaiman KAISAR ; Chang-Liang FU ; Rui-Yang CHEN
China Journal of Chinese Materia Medica 2006;31(2):114-116
OBJECTIVEStudy on the karyotypes analysis of Ferula fukanensis.
METHODThe young roots were treated with 0.000 2 mol x L(1) 8-Hydroxyquinoline for 3 h, carnoy's for 3 h, 1 mol x L(-1) HCl in 5 min,carbol fuchsin coloration for 2 min and the treated roots were utilizied to make the plate for observation.
RESULTThe fukanensis is diploid. The chromosome number of somatic cells was 2n = 22. The karyotype formula is 2n = 2x = 20 = 16m + 4sm. The 4th and 10th are submetacentric, and the others are metacentric.
CONCLUSIONThe karyotype of F. fukanensis belongs to "2A" type of stebbins', and it is a primitive species.
Centromere ; Chromosomes, Plant ; genetics ; Diploidy ; Ferula ; genetics ; Karyotyping ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; Seeds ; genetics
3.Screening and verification of proteins of Salvia miltiorrhiza polyphenol oxidase interaction.
Hai-Xing ZHANG ; Wang-Ke SHI ; Rong GUO ; Yue-Jin ZHANG ; Hong-Bo GUO
China Journal of Chinese Materia Medica 2020;45(11):2523-2532
Polyphenol oxidase(PPO) is an important antioxidant enzyme in plants. It has the functions of scavenging active oxygen and synthesizing phenols, lignin, and plant protection factors, and can enhance the plant's resistance to stress and resistance to pests and diseases. Our previous research found that Salvia miltiorrhiza PPO gene can positively regulate salvianolic acid B synthesis. In order to further explore the mechanism, a pGBKT7-PPO bait vector was constructed using the cloned S. miltiorrhiza polyphenol oxidase gene(SmPPO, GenBank accession number: KF712274.1), and verified that it had no self-activation and no toxicity. The titer of S. miltiorrhiza cDNA library constructed by our laboratory was 4.75 × 107 cfu·mL~(-1), which met the requirements for library construction. Through yeast two-hybrid test, 22 proteins that could interact with SmPPO were screened. Only yeast PAL1 and TAT interacted with SmPPO through yeast co-transformation verification. Further verification was performed by bimolecular fluorescence complementary detection(BiFC). Only TAT and SmPPO interacted, so it meant that TAT and SmPPO interacted. TAT and SmPPO were truncated according to the domain, respectively. The first 126 amino acids of SmPPO and tyrosine amino transferase(TAT) were obtained to interact on the cell membrane and chloroplast. SmPPO was obtained by subcellular localization test, which was mainly loca-lized on the nucleus and cell membrane; TAT was localized on the cell membrane. Real-time quantitative PCR results showed that the SmPPO gene was mainly expressed in roots and stems; the TAT gene was expressed in roots, and the expression level in stems and flowers was low. This article lays a solid foundation for the in-depth study of the molecular mechanism of the interaction of S. miltiorrhiza SmPPO and TAT to regulate the synthesis of phenolic substances.
Catechol Oxidase
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Gene Expression Regulation, Plant
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Gene Library
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Plant Proteins
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genetics
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Plant Roots
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Salvia miltiorrhiza
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genetics
4.Effects of density on growth and gene transcription characteristics of Rehmannia glutinosa.
Feng-Qing WANG ; Chao-Fei YANG ; Ming-Ming LI ; Xin ZUO ; Xu YANG ; Xin-Rong LI ; Li GU ; Jia-Fang DU ; Ming-Jie LI ; Hao WANG ; Zhong-Yi ZHANG
China Journal of Chinese Materia Medica 2021;46(17):4367-4379
The present study analyzed the effects of planting density on the development, quality, and gene transcription characte-ristics of Rehmannia glutinosa using 85-5 and J9 as materials with three planting densities of 5 000, 25 000, and 50 000 plants/Mu(1 Mu≈667 m~2). The agronomic characteristics of leaves and tuberous roots, the content of catalpol and acteoside, and the changes of gene expression were determined. The results showed that the leaf size, the diameter of tuberous root, leaf biomass, tuberous root number, and tuberous root biomass per plant at low density were significantly higher than those of medium and high densities. The content of catalpol and acteoside in leaves was higher at high density. The content of catalpol in tuberous roots was higher at low density, and the change trend was similar to that in leaves, while the content of acteoside in tuberous roots was higher at high density. Transcriptome analysis found that about 1/2 of the expansin genes could change regularly in response to density treatment, which was rela-ted to the development of tuberous roots. The change trend of the gene expression of multiple catalytic enzymes involved in the biosynthesis of catalpol and acteoside was consistent with that of their content, which was presumedly involved in the accumulation and regulation of density-responsive medicinal components. Based on the analysis of the development, medicinal components, and gene expression characteristics of R. glutinosa at different densities, this study is expected to provide an important basis for regulating the quality and yield of medicinal materials of R. glutinosa by managing the planting density.
Gene Expression Profiling
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Plant Leaves/genetics*
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Plant Roots/genetics*
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Rehmannia/genetics*
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Transcription, Genetic
5.Diversity of arbuscular mycorrhizal fungi of Panax quinquefolius cultivated in Shandong province.
Zhi-Fang RAN ; Xiao-Tong YANG ; Rui LI ; Jie ZHOU ; Yong-Qing ZHANG ; Lan-Ping GUO
China Journal of Chinese Materia Medica 2021;46(16):4103-4110
In this study, the colonization, diversity and relative abundance of arbuscular mycorrhizal fungi(AMF) in the roots of Panax quinquefolius in different habitats of Shandong province were analyzed by staining-microscopy and high-throughput sequencing. The data were analyzed by bioinformatics tools and statistical software. The results showed that the roots of P. quinquefolius in different habitats were colonized by AMF with different rates and intensities. The AMF in roots of P. quinquefolius belong to three genera, three families, three orders, one class and one phylum. At the level of order, the AMF mainly included Paraglomerales(52.48%), Glomerales(25.60%) and Archaeosporales(3.08%). At the level of family, the AMF were dominated by Paraglomeraceae(52.48%), Glomeraceae(18.94%) and Claroideoglomeraceae(3.05%). At the level of genus, Paraglomus(51.46%), Glomus(20.01%) and Claroideoglomus(3.52%) accounted for a large proportion, of which Paraglomus and Glomus were dominant. Cluster analysis showed that the AMF in roots of P. quinquefolius with close geographical locations could be clustered together. In this study, the diversity and dominant germplasm resources of AMF in roots of P. quinquefolius cultivated in the main producing areas were identified, which provi-ded basic data for revealing the quality formation mechanism of P. quinquefolius medicinal materials from the perspective of environment.
Fungi
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Glomeromycota
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Humans
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Mycorrhizae/genetics*
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Panax
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Plant Roots
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Soil Microbiology
6.Comparative analysis of active components and transcriptome between autotetraploid and diploid of Dendrobium huoshanense.
Cheng-Cai ZHANG ; Zhen GAO ; Li-Na LUO ; Hui-Hui LIANG ; Zeng-Xu XIANG
China Journal of Chinese Materia Medica 2020;45(23):5669-5676
In this study, the roots, stems and leaves of diploid and autotetraploid Dendrobium huoshanense were used as materials to compare their contents of polysaccharides and alkaloids, and the transcriptome sequencing analysis was carried out. The results showed that the contents of polysaccharides and alkaloids in the roots, stems and leaves of tetraploid were 7.6%, 34.5%, 17.2%, 0.01%, 0.024% and 0.035% higher than those of diploid D. huoshanense, respectively. The contents of active components in different tissues were significantly different. There were 3 687 differentially expressed genes in diploid and tetraploid D. huoshanense, of which 2 346 genes were up-regulated and 1 341 down regulated. Go functional analysis showed that these genes were mainly involved in growth and development, stress resistance and other related functions. KEGG pathway analysis showed that most of the differential genes were concentrated in the processes of carbon metabolism, signal transduction, carbohydrate metabolism, amino acid metabolism and energy metabolism. The differential expression of key genes involved in the metabolism of polysaccharides, terpenes and polyketones, amino acid metabolism, hormone synthesis and signal transduction in diploid and tetraploid plants may be the main reason for the high energy content, the increase of active components and the growth potential of tetraploid plants.
Alkaloids
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Dendrobium/genetics*
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Diploidy
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Plant Roots
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Polysaccharides
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Transcriptome
7.Transcriptome analysis of Platycodon grandiflorum at different growth years and discovery of key genes for platycodin biosynthesis.
Jian ZHANG ; Yuan PAN ; Yu WANG ; Da-Xia CHEN
China Journal of Chinese Materia Medica 2021;46(6):1386-1392
Platycodon grandiflorum is a medicinal and edible medicinal material. Our study is aimed to explore the differences in the gene expression of P. grandiflorum in different growth years, and the expression rules of key genes in the biosynthesis of the main active substances of P. grandiflorum. Illumina Hiseq 4000 sequencing platform was used to sequence the transcriptome of P. grandiflorum in different years. Then, 59 654 unigenes were obtained through filtering, assembly, splicing and bioinformatics analysis of the sequencing data, of which 1 671 unigenes were differentially expressed between at least two samples. The results of cluster analysis showed that there was a great difference in the gene expression of P. grandiflorum from one-year-old to two/three-year-old. There were 1 128 different genes between one-and three-year old P. grandiflorum, and only 57 different genes between two-and three-year-old P. grandiflorum. KEGG enrichment results showed that the differential genes of P. grandiflorum in different years were mainly concentra-ted in the biosynthesis of sesquiterpenes and triterpenes, and the biosynthesis of terpenoid skeletons. In the triterpenoid biosynthesis-related pathways, a total of 15 unigenes were identified, involving 5 enzymes. The expression levels of ACAT, HMGR, FDFT1, SQLE decreased with the increase of the growth year of P. grandiflorum. The expression of HMGS was the highest in the one-year-old P. grandiflorum, followed by the three-year-old sample. This study provides useful data for the development of P. grandiflorum, and also provides a basis for the study of related genes in the biosynthetic pathway of platycodin.
Gene Expression Profiling
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Plant Roots
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Platycodon/genetics*
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Saponins
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Transcriptome
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Triterpenes
8.Genome-wide identification of BvHAK gene family in sugar beet (Beta vulgaris) and their expression analysis under salt treatments.
Xiaohan YANG ; Guoqiang WU ; Ming WEI ; Beichen WANG
Chinese Journal of Biotechnology 2022;38(10):3773-3789
High-affinity K+ transporter (HAK) is one of the most important K+ transporter families in plants and plays an important role in plant K+ uptake and transport. To explore the biological functions and gene expression patterns of the HAK gene family members in sugar beet (Beta vulgaris), physicochemical properties, the gene structure, chromosomal location, phylogenetic evolution, conserved motifs, three-dimensional structure, interaction network, cis-acting elements of promoter of BvHAKs were predicted by bioinformatic analysis, and their expression levels in different tissues of sugar beet under salt stress were analyzed by qRT-PCR. A total of 10 BvHAK genes were identified in the sugar beet genome. They contained 8-10 exons and 7-9 introns. The average number of amino acids was 778.30, the average molecular weight was 88.31 kDa, and the isoelectric point was 5.38-9.41. The BvHAK proteins contained 11-14 transmembrane regions. BvHAK4, -5, -7 and -13 were localized on plasma membrane, while others were localized on tonoplast. Phylogenetic analysis showed that HAK in higher plants can be divided into five clusters, namely cluster Ⅰ, Ⅱ, Ⅲ, Ⅳ, and Ⅴ, among which the members of cluster Ⅱ can be divided into three subclusters, including Ⅱa, Ⅱb, and Ⅱc. The BvHAK gene family members were distributed in cluster Ⅰ-Ⅳ with 1, 6, 1, and 2 members, respectively. The promoter of BvHAK gene family mainly contained stress responsive elements, hormone responsive elements, and growth and development responsive elements. The expression pattern of the BvHAK genes were further analyzed in different tissues of sugar beet upon salt treatment, and found that 50 and 100 mmol/L NaCl significantly induced the expression of the BvHAK genes in both shoots and roots. High salt (150 mmol/L) treatment clearly down-regulated their expression levels in shoots, but not in roots. These results suggested that the BvHAK gene family plays important roles in the response of sugar beet to salt stress.
Beta vulgaris/genetics*
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Gene Expression Regulation, Plant
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Phylogeny
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Plant Roots
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Sugars/metabolism*
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Plant Proteins/metabolism*
9.Studies on chromosome numbers of Salvia miltiorrhiza, S. flava and S. evansiana.
Hong-xia ZHAO ; Li ZHANG ; Xing FAN ; Rui-wu YANG ; Chun-bang DING ; Yong-hong ZHOU
China Journal of Chinese Materia Medica 2006;31(22):1847-1849
OBJECTIVETo study the numbers of chromosome in Chinese herb Salvia miltiorrhiza from 7 provinces in China, and S. flava as well as S. evansiana from Yunnan province in China.
METHODThe young root was treated with the mixture of ice and water for 24 h, fixed with Carony's fixative for 6-12 h. After differentiating for 10-12 min with 1 mol x L(-1) hydrochloric acid at 60 'C and staining with carbol fuchsin,the section was observed under microscope.
RESULTChromosome numbers of S. miltiorrhiza and S.flava were 2n = 2x = 16. The numbers of S. evansiana were 2n = 4x = 32. The basic numbers of the chromosomes were x = 8. And tetraploids were observed in S. miltiorrhiza from Sichuan provices and Shandong provices.
CONCLUSIONThe basic number of the chromosomes are x = 8. The chromosome numbers of S. miltiorrhiza, S.flava and S. evansiana are 16,16 and 32 respectively. As the chromosomes are the small or micro-small ones, it is difficult to use them for karyotype.
Chromosomes, Plant ; genetics ; Diploidy ; Plant Roots ; genetics ; Plants, Medicinal ; genetics ; Polyploidy ; Salvia miltiorrhiza ; classification ; genetics ; Species Specificity
10.De novo sequencing and analysis of root transcriptome to reveal regulation of gene expression by moderate drought stress in Glycyrrhiza uralensis.
Chun-rong ZHANG ; Xue-yu SANG ; Meng QU ; Xiao-min TANG ; Xuan-xuan CHENG ; Li-ming PAN ; Quan YANG
China Journal of Chinese Materia Medica 2015;40(24):4817-4823
Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts
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Gene Expression Regulation, Plant
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Glycyrrhiza uralensis
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genetics
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Plant Roots
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Sequence Analysis, DNA
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Stress, Physiological
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Transcriptome