1.Mechanisms of alternative splicing in regulating plant flowering: a review.
Huanhuan LU ; Qinlin DENG ; Mengdan WU ; Zhimin WANG ; Dayong WEI ; Hebing WANG ; Huafeng XIANG ; Hongcheng ZHANG ; Qinglin TANG
Chinese Journal of Biotechnology 2021;37(9):2991-3004
Flowering is a critical transitional stage during plant growth and development, and is closely related to seed production and crop yield. The flowering transition is regulated by complex genetic networks, whereas many flowering-related genes generate multiple transcripts through alternative splicing to regulate flowering time. This paper summarizes the molecular mechanisms of alternative splicing in regulating plant flowering from several perspectives, future research directions are also envisioned.
Alternative Splicing/genetics*
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Arabidopsis/metabolism*
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Arabidopsis Proteins/genetics*
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Flowers/genetics*
2.Expression and function analysis of FaCO gene in Festuca arundinacea.
Xi CHEN ; Ying CHEN ; Xiaoxia LIU ; Jianhong SHU ; Xiaoli WANG ; Degang ZHAO
Chinese Journal of Biotechnology 2021;37(4):1324-1333
Photoperiod plays an important role in transformation from vegetative growth to reproductive growth in plants. CONSTANS (CO), as a unique gene in the photoperiod pathway, responds to changes of day length to initiate flowering in the plant. In this study, the expression level of FaCONSTANS (FaCO) gene under long-day, short-day, continuous light and continuous darkness conditions was analyzed by real-time quantitative PCR. We constructed the over-expression vector p1300-FaCO and infected into Arabidopsis thaliana by Agrobacterium-mediated method. We constructed the silencing vector p1300-FaCO-RNAi and infected into Festuca arundinacea by Agrobacterium-mediated method. The expression of FaCO gene was regulated by photoperiod. The over-expression of FaCO promoted flowering in wild type of Arabidopsis thaliana under long day condition and rescued the late flowering phenotype in co-2 mutant of Arabidopsis thaliana. Silencing FaCO gene in Festuca arundinacea by RNAi showed late-flowering phenotype or always kept in the vegetative growth stage. Our understanding the function of FaCO in flowering regulation will help further understand biological function of this gene in Festuca arundinacea.
Arabidopsis/metabolism*
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Arabidopsis Proteins/genetics*
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Festuca/metabolism*
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Flowers/genetics*
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Gene Expression Regulation, Plant
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Photoperiod
3.CLE42 binding induces PXL2 interaction with SERK2.
Shulin MOU ; Xiaoxiao ZHANG ; Zhifu HAN ; Jiawei WANG ; Xinqi GONG ; Jijie CHAI
Protein & Cell 2017;8(8):612-617
Arabidopsis
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chemistry
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genetics
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metabolism
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Arabidopsis Proteins
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chemistry
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genetics
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metabolism
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Crystallography, X-Ray
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Intercellular Signaling Peptides and Proteins
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chemistry
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genetics
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metabolism
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Protein Conformation
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Protein-Serine-Threonine Kinases
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chemistry
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genetics
;
metabolism
4.Identification of glucose-responsive elements in the promoter of UDP-L-rhamnose biosynthesis gene RHM1 in Arabidopsis thaliana.
Qinmei JI ; Jinfeng WANG ; Ling JIANG ; Songdong SHEN ; Yunliu FAN ; Chunyi ZHANG
Chinese Journal of Biotechnology 2008;24(9):1531-1537
In plants, UDP-L-rhamnose is one of the major components of cell wall skeleton. Rhamnose synthase plays a key role in rhamnose synthesis which converts UDP-D-glucose into UDP-L-rhamnose in plants. In this study, we isolated the 1058 bp promoter region of the rhamnose synthase gene AtRHM1 from Arabidopsis genome by PCR, and created a series of deletions of AtRHM1 promoter ranging from -931 bp to +127 bp. The full length of the promoter and its deletion derivatives fused with GUS reporter gene were introduced into wild-type Arabidopsis by Agrobacterium-mediated transformation respectively. The GUS staining and GUS enzymatic activity assay showed that the expression of AtRHM1 is induced at transcriptional level by glucose and the regulatory elements involved in the glucose response are located in the region of -931 bp - -752 bp which contains three G-box motifs.
Arabidopsis
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genetics
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Arabidopsis Proteins
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genetics
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Glucosyltransferases
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genetics
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Plants, Genetically Modified
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genetics
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Promoter Regions, Genetic
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Uridine Diphosphate Glucose
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genetics
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metabolism
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Uridine Diphosphate Sugars
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genetics
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metabolism
5.Prokaryotic expression, purification and functional identification of epidermal pattern factors in Arabidopsis thaliana.
Zhuping JIN ; Cheng LI ; Lei WANG ; Yanxi PEI
Chinese Journal of Biotechnology 2020;36(4):792-800
Stomatal density is important for crop yield. In this paper, we studied the epidermal pattern factors (EPFs) related to stomatal development. Prokaryotic expression vectors were constructed to obtain EPFs. Then the relationship between EPFs and hydrogen sulfide (H2S) was established. First, AtEPF1, AtEPF2 and AtEPFL9 were cloned and constructed to pET28a vectors. Then recombinant plasmids pET28a-AtEPF1, pET28a-AtEPF2 and pET28a-AtEPFL9 were digested and sequenced, showing successful construction. Finally, they were transformed into E. coli BL21(DE3) separately and induced to express by isopropyl β-D-galactoside (IPTG). The optimized expression conditions including IPTG concentration (0.5, 0.3 and 0.05 mmol/L), temperature (28 °C, 28 °C and 16 °C) and induction time (16 h, 16 h and 20 h) were obtained. The bands of purified proteins were about 18 kDa, 19 kDa and 14.5 kDa, respectively. In order to identify their function, the purified AtEPF2 and AtEPFL9 were presented to Arabidopsis thaliana seedlings. Interestingly, the H2S production rate decreased or increased compared with the control, showing significant differences. That is, EPFs affected the production of endogenous H2S in plants. These results provide a foundation for further study of the relationship between H2S and EPFs on stomatal development, but also a possible way to increase the yield or enhance the stress resistance.
Arabidopsis
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genetics
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metabolism
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Arabidopsis Proteins
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genetics
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isolation & purification
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metabolism
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Escherichia coli
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genetics
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Genetic Vectors
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genetics
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Hydrogen Sulfide
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metabolism
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Plasmids
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genetics
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Seedlings
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metabolism
6.Subcellular localization and resistance to Gibberella fujikuroi of AtELHYPRP2 in transgenic tobacco.
Qiuxia CHAI ; Benchang LI ; Ziqin XU
Chinese Journal of Biotechnology 2014;30(3):472-484
The subcellular localization and the resistance to fungal pathogen Gibberella fujikuroi of the protein encoded by Arabidopsis AtELHYPRP2 (EARLI1-LIKE HYBRID PROLINE-RICH PROTEIN 2, AT4G12500) were investigated using transgenic tobacco plants. The coding sequence of AtELHYPRP2 was amplified from genomic DNA of Col-0 ecotype. After restriction digestion, the PCR fragment was ligated into pCAMBIA1302 to produce a fusion expression vector, pCAMBIA1302-AtELHYPRP2-GFP. Then the recombinant plasmid was introduced into Agrobacterium tumefaciens strain LBA4404 and transgenic tobacco plants were regenerated and selected via leaf disc transformation method. RT-PCR and Western blotting analyses showed that AtELHYPRP2 expressed effectively in transgenic tobacco plants. Observation under laser confocal microscopy revealed that the green fluorescence of AtELHYPRP2-GFP fusion protein could overlap with the red fluorescence came from propidium iodide staining, indicating AtELHYPRP2 is localized to cell surface. Antimicrobial experiments exhibited that the constitutive expression of AtELHYPRP2 could enhance the resistance of tobacco to fungal pathogen G. fujikuroi and the infection sites could accumulate H2O2 obviously. The basal expression levels of PR1 and the systemic expression levels of PR1 and PR5 in transgenic tobacco plants were higher than that of the wild-type plants, suggesting AtELHYPRP2 may play a role in systemic acquired resistance.
Agrobacterium tumefaciens
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Arabidopsis
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Arabidopsis Proteins
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genetics
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Disease Resistance
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Gibberella
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pathogenicity
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Hydrogen Peroxide
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Plants, Genetically Modified
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microbiology
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Recombinant Fusion Proteins
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genetics
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Tobacco
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genetics
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microbiology
7.Cloning of AhWRKY33 from Asarum heterotropoides and its genetic transformation in Arabidopsis.
Yong-Cheng YANG ; Xiao-Han WANG ; Yan-Fang WANG ; Xiu-Yan LIU ; Shi-Hai YANG
China Journal of Chinese Materia Medica 2020;45(13):3112-3119
The WRKY family genes, which play an important role in plant morphogenesis and stress response, were selected based on the data of the full-length transcriptome of Asarum heterotropoides. Using AtWRKY33, which regulates the synthesis of the camalexin in the model plant Arabidopsis to compare homologous genes in A. heterotropoides, primers were designed to amplify the open reading frame(ORF) fragment of AhWRKY33 gene by RT-PCR using total RNA of A. heterotropoides leaves as template. Real-time PCR results showed that there was a significant difference between the aerial part and the underground part of A. heterotropoides, the toxic aristolochic acid content is highly expressed in the leaves higher than the root. After verification, the WRKY33 gene of A. heterotropoides is ORF long 1 686 bp, encoding 561 amino acids.AhWRKY33 had two conserved WRKYGQK domains. According to the classical classification, it belongs to group Ⅰ WRKY transcription factor. A. heterotropoides WRKY33 had some homology with amino acids of other species. The study successfully constructed the plant eukaryotic expression vector PHG-AhWRKY33 and transformed Arabidopsis thaliana, the transgenic Arabidopsis was obtained by PCR detection and hygromycin resistant plate screening. It found that the germination of transgenic Arabidopsis seeds was accelerated and the stress resistance was increased. It laid a foundation for further analysis of WRKY transcription factor in the growth and development of A. heterotropoides and the synthesis of secondary metabolites.
Arabidopsis
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genetics
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Arabidopsis Proteins
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genetics
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Asarum
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Cloning, Molecular
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Gene Expression Regulation, Plant
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Plant Leaves
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Plant Proteins
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genetics
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Transcription Factors
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Transformation, Genetic
8.Role of plant autophagy in stress response.
Shaojie HAN ; Bingjie YU ; Yan WANG ; Yule LIU
Protein & Cell 2011;2(10):784-791
Autophagy is a conserved pathway for the bulk degradation of cytoplasmic components in all eukaryotes. This process plays a critical role in the adaptation of plants to drastic changing environmental stresses such as starvation, oxidative stress, drought, salt, and pathogen invasion. This paper summarizes the current knowledge about the mechanism and roles of plant autophagy in various plant stress responses.
Adaptation, Physiological
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Arabidopsis
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genetics
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physiology
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Arabidopsis Proteins
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genetics
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metabolism
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Autophagy
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genetics
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Disease Resistance
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Plant Diseases
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immunology
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Saccharomyces cerevisiae
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genetics
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Sequence Homology
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Stress, Physiological
9.Cloning and catalytic analysis of Isatis indigotica chalcone isomerase in vitro.
Ke-Ke ZHANG ; Shu-Fu SUN ; Yu-Ping TAN ; Zhao-Yang XU ; Yin-Yin JIANG ; Jian YANG ; Da-Yong LI ; Jin-Fu TANG
China Journal of Chinese Materia Medica 2023;48(6):1510-1517
Chalcone isomerase is a key rate-limiting enzyme in the biosynthesis of flavonoids in higher plants, which determines the production of flavonoids in plants. In this study, RNA was extracted from different parts of Isatis indigotica and reverse-transcribed into cDNA. Specific primers with enzyme restriction sites were designed, and a chalcone isomerase gene was cloned from I. indigotica, named IiCHI. IiCHI was 756 bp in length, containing a complete open reading frame and encoding 251 amino acids. Homology analysis showed that IiCHI was closely related to CHI protein of Arabidopsis thaliana and had typical active sites of chalcone isomerase. Phylogenetic tree analysis showed that IiCHI was classified into type Ⅰ CHI clade. Recombinant prokaryotic expression vector pET28a-IiCHI was constructed and purified to obtain IiCHI recombinant protein. In vitro enzymatic analysis showed that the IiCHI protein could convert naringenin chalcone into naringenin, but could not catalyze the production of liquiritigenin by isoliquiritigenin. The results of real-time quantitative polymerase chain reaction(qPCR) showed that the expression level of IiCHI in the aboveground parts was higher than that in the underground parts and the expression level was the highest in the flowers of the aboveground parts, followed by leaves and stems, and no expression was observed in the roots and rhizomes of the underground parts. This study has confirmed the function of chalcone isomerase in I. indigotica and provided references for the biosynthesis of flavonoid components.
Isatis/genetics*
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Plant Proteins/metabolism*
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Phylogeny
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Arabidopsis/genetics*
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Flavonoids
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Cloning, Molecular
10.Progress in the plant GH3 gene family.
Tao SUN ; Tuanyao CHAI ; Geyu LIU ; Yuxiu ZHANG
Chinese Journal of Biotechnology 2008;24(11):1860-1866
Phytohormone auxins play important roles in plant growth and development. The primary auxin-response genes can be classified into three major groups: Aux/IAAs, SAURs and GH3s. Significant progress has been made in understanding these gene families by approaches of the functional genomics, molecular genetics and molecular biology. In this review, we focused on the structures, functions and models of the expressional regulation of plant GH3 genes. The interactions in the signal transduction pathways between auxins and other signals mediated by the GH3 genes, the relationship between the GH3 genes and the stress adaptation responses of plants are emphasized.
Arabidopsis Proteins
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genetics
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Gene Expression Regulation, Plant
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Genes, Plant
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Ligases
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genetics
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Multigene Family
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Plant Growth Regulators
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genetics
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Soybean Proteins
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genetics