1.Effect of short-day treatment on expression of CO gene in Chrysanthemum indicum.
Dan-Dan CHEN ; Qing-Jun ZOU ; Qiao-Sheng GUO ; Tao WANG
China Journal of Chinese Materia Medica 2019;44(4):648-653
This experiment studied the expression pattern of key gene CO in the photoperiod of Chrysanthemum indicum. The CDS sequence of the Ch. indicum CO gene was cloned by RT-PCR. The open reading frame was 1 380 bp in length and encoded 459 amino acids. The bioinformatics analysis results showed that the Ch. indicum CO had higher homology with Ch. lavandulifolium and Artemisia annua,and the CO was more conservative in the same family. The molecular weight of the predicted protein encode by CO is 52. 04 k Da,the p I is 4. 81,the α-helix structure accounted for 17. 65%,the random coil accounted for 76. 69%,the extension chain accounted for 5. 66%,there are no β-fold and signal peptide. The experimental results showed that short-day treatment could increase the expression level of CO gene in Ch. indicum and induce its flowering. The results of qRT-PCR showed that the relative expression of CO gene in different tissues and different treatment periods of Ch. indicum was significantly different. In this paper,we studied the effect of short-day treatment on the expression of key genes in the flowering cycle of Ch. indicum,providing a basis for photoperiod regulation and harvesting period of Ch.indicum.
Chrysanthemum
;
Gene Expression Regulation, Plant
2.Transcriptome analysis of seed embryo in dormancy and dormancy release state of Thesium chinense.
Cheng-Cai ZHANG ; Zhen GAO ; Li-Na LUO ; Zi-Xuan ZHANG ; Jiang WANG ; Fu-Hua LU ; Zeng-Xu XIANG
China Journal of Chinese Materia Medica 2020;45(16):3837-3843
We used exogenous GA_3 to break the seed dormancy of Thesium chinense. We used high-throughput sequencing technology was used to sequence the transcriptome of dormant seed embryos and dormancy breaking seed embryos of Th. chinense, and the data was analyzed bioinformatically and systematically. The results showed that exogenous GA_3 could effectively break the seed dormancy of Th. chinense; 73 794 up-regulated genes and 42 776 down regulated genes were obtained by transcriptome sequencing; 116 570 diffe-rential genes were annotated by GO function to GO items such as metabolism process, cell process, cell, cell component, binding and catalytic activity. A total of 133 metabolic pathways were found by Pathway analysis of 26 508 differentially expressed genes. In the process of dormancy release, DEGs were mainly enriched in translation, carbohydrate metabolism, folding, classification, degradation and amino acid metabolism. Based on the annotation results in KEGG database, 20 metabolic pathways related to dormancy release were found. Dormancy release of Th. chinense seeds is a complex biological process, including cell morphology construction, secondary metabolite synthesis, sugar metabolism and plant signal transduction, among which plant hormone signal transduction is one of the key factors to regulate dormancy release. The results of qRT-PCR showed that the sequencing results were consistent with the actual results.
Gene Expression Profiling
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Gene Expression Regulation, Plant
;
Plant Dormancy
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Plant Growth Regulators
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Santalaceae
;
Seeds
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Transcriptome
3.Ectopic expression of archaeal TRAM-encoding genes in rice improves its drought-tolerance.
Wei CHEN ; Huali LI ; Jinlong QIU
Chinese Journal of Biotechnology 2019;35(9):1676-1685
Drought stress affects the growth and development of rice, resulting in severe loss in yield and quality. Ectopic expression of the bacterial RNA chaperone, cold shock protein (Csp), can improve rice drought tolerance. Archaeal TRAM (TRM2 and MiaB) proteins have similar structure and biochemical functions as bacterial Csp. Moreover, DNA replication, transcription and translation of archaea are more similar to those in eukaryotes. To test if archaeal RNA chaperones could confer plant drought tolerance, we selected two TRAM proteins, Mpsy_3066 and Mpsy_0643, from a cold-adaptive methanogenic archaea Methanolobus psychrophilus R15 to study. We overexpressed the TRAM proteins in rice and performed drought treatment at seedling and adult stage. The results showed that overexpression both TRAM proteins could significantly improve the tolerance of rice to drought stress. We further demonstrated in rice protoplasts that the TRAMs could abolish misfolded RNA secondary structure and improve translation efficiency, which might explain how TRAMs improve drought tolerance transgenic rice. Our work supports that ectopic expression of archaeal TRAMs effectively improve drought tolerance in rice.
Droughts
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Ectopic Gene Expression
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Gene Expression Regulation, Plant
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Oryza
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Plant Proteins
;
Plants, Genetically Modified
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Stress, Physiological
4.Differences between male and female leaves of Schisandra sphenanthera: based on RNA-Seq.
Jie LI ; Jia-Qi GAO ; Tie-Gui NAN ; Jun-Hui ZHOU ; Zhong-Yi HUA ; Jian YANG ; Chao JIANG ; Lu-Qi HUANG
China Journal of Chinese Materia Medica 2021;46(22):5797-5803
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.
Abscisic Acid
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Gene Expression Profiling
;
Gene Expression Regulation, Plant
;
Plant Leaves/genetics*
;
RNA-Seq
;
Schisandra
;
Transcriptome
5.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
;
Gene Library
;
Plant Proteins
;
genetics
;
Plant Roots
;
Salvia miltiorrhiza
;
genetics
6.Application of virus-induced gene silencing technology to investigate the phytochrome metabolism mechanism: a review.
Duo PAN ; Songyue ZHANG ; Fangyi LIU ; Qingyin TIAN ; Xiulian YANG ; Lianggui WANG ; Yuanzheng YUE
Chinese Journal of Biotechnology 2023;39(7):2579-2599
Color is an important indicator for evaluating the ornamental traits of horticultural plants, and plant pigments is a key factor affecting the color phenotype of plants. Plant pigments and their metabolites play important roles in color formation of ornamental organs, regulation of plant growth and development, and response to adversity stress. It has therefore became a hot topic in the field of plant research. Virus-induced gene silencing (VIGS) is a vital genomics tool that specifically reduces host endogenous gene expression utilizing plant homology-dependent defense mechanisms. In addition, VIGS enables characterization of gene function by rapidly inducing the gene-silencing phenotypes in plants. It provides an efficient and feasible alternative for verifying gene function in plant species lacking genetic transformation systems. This paper reviews the current status of the application of VIGS technology in the biosynthesis, degradation and regulatory mechanisms of plant pigments. Moreover, this review discusses the potential and future prospects of VIGS technology in exploring the regulatory mechanisms of plant pigments, with the aim to further our understandings of the metabolic processes and regulatory mechanisms of different plant pigments as well as improving plant color traits.
Plant Viruses/genetics*
;
Plants/genetics*
;
Gene Silencing
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Plant Development
;
Gene Expression Regulation, Plant
;
Genetic Vectors
7.Identification of terpene synthase gene family members in Rhododendron and its relationship with terpenoid metabolism.
Guoxia YANG ; Baoxin JIANG ; Fan HE ; Sijia LÜ ; Dongbin LI ; Yonghong JIA ; Ping ZHU ; Xiaohong XIE ; Yueyan WU
Chinese Journal of Biotechnology 2022;38(10):3740-3756
Terpene synthase (TPS) plays important roles in the synthesis of terpenoids which are the main fragrances in Rhododendron flowers. To understand the function of TPS genes in terpenoid metabolism in relation to flower aroma formation, we identified all TPS gene family members in Rhododendron by analyzing its genome database. We then used a transcriptomic approach to analyze the differential gene expression patterns of TPS gene family members in the scented flower Rhododendron fortunei compared to the non-scented flower Rhododendron 'Nova Zembla'. The contents of terpenoid compounds in petals of the above two Rhododendron species at different developmental stages were also measured by using qRT-PCR and head space-solid phase micro-extraction combined with gas chromatography-mass spectrometry. Our results showed that a total of 47 RsTPS members, with individual lengths ranged from 591 to 2 634 bp, were identified in the Rhododendron genome. The number of exons in RsTPS gene ranged from 3 to 12, while the length of each protein encoded ranged from 196 to 877 amino acids. Members of the RsTPS family are mainly distributed in the chloroplast and cytoplasm. Phylogenetic analysis showed that RsTPS genes can be clustered into 5 subgroups. Seven gene family members can be functionally annotated as TPS gene family since they were temporally and spatially expressed as shown in the transcriptome data. Notably, TPS1, TPS10, TPS12 and TPS13 in Rhododendron fortunei were expressed highly in flower buds reached the peak in the full blossoming. Correlation analysis between gene expression levels and terpenoid content indicates that the expression levels of TPS1, TPS4, TPS9, TPS10, TPS12 and TPS13 were positively correlated with the content of terpenoids in the petals of R. fortunei at all flower developmental stages, suggesting that these six genes might be involved in the aroma formation in R. fortunei.
Rhododendron/metabolism*
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Phylogeny
;
Terpenes/metabolism*
;
Family
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Gene Expression Regulation, Plant
8.Advances in the physiological functions of plant lipids in response to stresses.
Junyu LIU ; Fan YANG ; Shuang MAO ; Shuxin LI ; Haijiao LIN ; Xiufeng YAN ; Jixiang LIN
Chinese Journal of Biotechnology 2021;37(8):2658-2667
Lipids are important components of living organisms that participate in and regulate a variety of life activities. Lipids in plants also play important physiological functions in response to a variety of abiotic stresses (e.g. salt stress, drought stress, temperature stress). However, most research on lipids focused on animal cells and medical fields, while the functions of lipids in plants were overlooked. With the rapid development of "omics" technologies and biotechnology, the lipidomics has received much attention in recent years because it can reveal the composition and function of lipids in a deep and comprehensive way. This review summarizes the recent advances in the functions and classification of lipids, the development of lipidomics technology, and the responses of plant lipids against drought stress, salt stress and temperature stress. In addition, challenges and prospects were proposed for future lipidomics research and further exploration of the physiological functions of lipids in plant stress resistance.
Droughts
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Gene Expression Regulation, Plant
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Lipids
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Plants
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Stress, Physiological
9.Studies on functional genes of Chinese herbal medicine.
Xiao-yong WEI ; Hua FANG ; Jie-feng LI
Chinese Journal of Integrated Traditional and Western Medicine 2005;25(2):181-185
In order to establish the foundation for modernization of Chinese herbal medicine, discovery of the functional genes related with the bioactive components of Chinese herbs and the rules of their expression was progressed for determining the regulatory mechanism, obtaining the core metabolic pathway and key regulatory factors of the medicinal components synthesis. In this article, the studies about plant's functional genes at home and abroad was introduced in three aspects: the pattern plants (rice, thaliana, etc.), the main technologic strategy (gene expression differentia, sequential tag of gene expression, DNA microarray technique and gene expression sequential analysis) and bio-informative method (comparative genomics). The current status of researches involving biosynthesis genes of flavonoids compounds and taxol, and the plant P450 genes were introduced. And the study of dendrobium functional genes conducted in authors' lab was also reviewed.
Drugs, Chinese Herbal
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Gene Expression Profiling
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Gene Expression Regulation, Plant
;
Genome, Plant
;
Plant Proteins
;
genetics
;
Plants, Medicinal
;
genetics
10.Transcriptome-wide identification and expression pattern analysis for Dof gene family in Panax ginseng from Jilin.
Kang-Yu WANG ; Yan-Xiu GUO ; Chong MA ; Ming-Zhu ZHAO ; Feng-Hua LI ; Mei-Ping ZHANG ; Yi WANG
China Journal of Chinese Materia Medica 2022;47(1):62-71
Dof(DNA binding with one finger), a unique class of transcription factors in plants, play an important role in seed development, tissue differentiation, and metabolic regulation. To identify the number and function of Dof gene family members in Panax ginseng, this study identified the members of Dof gene family in P. ginseng and systematically analyzed their structures, evolution, functional differentiation, expression patterns, and interactions using bioinformatics methods at the transcriptome level. At the same time, the association analysis of Dof genes from P. ginseng with key enzyme genes for ginsenoside synthesis was carried out to screen the candidate PgDof genes involved in the regulation of ginsenoside biosynthesis. The results showed that there were 54 genes belonging to the Dof gene family in P. ginseng from Jilin. All PgDof genes had Zf-Dof conserved motifs, implying that they were evolutionarily conserved and could be divided into five groups. Expression pattern analysis confirmed that the expression of PgDof gene family members in different tissues, different year-old P. ginseng, and different farm varieties varied significantly. Simultaneously, as revealed by "gene-saponin content" and "gene-gene" linkage analysis, an important candidate PgDof14-1 gene involved in the regulation of ginsenoside biosynthesis was obtained. From the established genetic transformation system of this gene in the hairy roots of P. ginseng, a positive hairy root clone was determined. This study has laid a theoretical foundation for the study of Dof gene family in P. ginseng.
Gene Expression Profiling/methods*
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Gene Expression Regulation, Plant
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Ginsenosides
;
Panax
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Plant Proteins/metabolism*
;
Plant Roots/metabolism*
;
Transcriptome