1.Construction and sequence analysis of a drought-induced full-length cDNA library from Ammopiptanthus mongolicus.
Qingfang LIN ; Xuefeng WANG ; Jiyuan LI ; Hongbin ZHAO ; Maoyan WANG
Chinese Journal of Biotechnology 2012;28(1):86-95
Ammopiptanthus mongolicus shows very strong resistance to severe environments. To isolate drought-resistant genes and elucidate drought-resistant molecular mechanisms of the plant, we constructed a drought-induced full-length cDNA library using SMART (Switching mechanism at 5'-end of RNA transcript) technique. The phage titer of the unamplified library was 1.6 x 10(7) PFU/mL; the recombination percentage was 97.7%; and the sizes of most cloned cDNA fragments were around 1 kb. Three thousand positive clones were randomly selected and sequenced from their 5' ends, and a total of 1 450 Unigenes were identified. By Blast searches against the Nt, Nr and Swissprot databases, we found that 919 Unigenes (amount to 63.4%) showed significant similarity to the annotated genes, and the remaining 531 Unigenes (amount to 36.6%) represented novel genes without any annotation. Among the functional categories of the GO (Gene Ontology) classification, the terms related to physiological process, cellular process, binding, catalytic activity and cellular components were dominant. The next abundant terms were for organelle, protein complex, transporter activity and structural molecule activity. In addition, there were a significant proportion of the terms involved in stimulus response, gene expression regulation, regulation of physiological and biochemical processes and signal transduction. Many of the annotated Unigenes were found to be related to plant resistance to abiotic stresses, and expression analyses of 6 out of these genes by semi-quantitive RT-PCR confirmed their involvements in the response of A. mongolicus to drought stress. These results laid a foundation for the expression profile analysis and the cloning and characterization of drought-resistant genes from the plant in the future.
Droughts
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Fabaceae
;
genetics
;
Gene Expression Regulation, Plant
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Gene Library
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Genotype
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Sequence Analysis, DNA
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Stress, Physiological
;
genetics
2.Identification of soybean GolS gene family and analysis of expression patterns under salt and drought stresses.
Dan LIU ; Keai WANG ; Peng NI ; Qiuyan WANG ; Kang ZHU ; Wenliang WEI
Chinese Journal of Biotechnology 2022;38(10):3757-3772
Galactinol synthase (GolS) is a key enzyme in the biosynthetic pathway of raffinose family oligosaccharides (RFOs) and plays an important role in plant responses to abiotic stresses. However, the molecular characteristics of the GolS family members in soybean was not well-known. In this study, six members of GmGolS gene family were genome-widely identified, and their physicochemical properties, chromosomal localization, evolutionary relationship, gene structure, conserved motifs, secondary structure, tertiary structure, tissue-specific expression patterns and the expression levels under salt and drought stresses were analyzed. The results showed that six soybean GolS genes were unevenly distributed on four chromosomes, the range of the isoelectric points of six GmGolS proteins was 5.45-6.08, the molecular weight range was 37 567.07-38 817.59 Da, and the number of amino acids was 324-339 aa. The results of subcellular localization showed that 4 proteins were located in the chloroplast, and 2 proteins in the cytoplasm. Phylogenetic tree analysis showed that the members of the soybean GolS gene family were closely adjacent to each other, and were evolutionarily conservative. Six gene members contain 3 or 4 exons. Prediction of secondary and tertiary structures showed that the spatial structure of proteins of all family members was mainly composed of α-helix and random coil structure, with less β-turn and extended chain structure. Tissue-specific expression analysis showed that six GmGolS members expressed to variable degrees in seeds, roots, root hairs, flowers, stems, pods, nodules and leaves. Expression analysis based on qRT-PCR showed that all GmGolS genes showed different degrees of up-regulated expression under salt and drought treatment, indicating that these genes may be related to the response of plants to salt-tolerance and drought-resistance. These results may facilitate subsequent functional analysis of soybean GolS genes.
Droughts
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Soybeans/genetics*
;
Phylogeny
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Plant Proteins/metabolism*
;
Stress, Physiological/genetics*
;
Plants/metabolism*
;
Gene Expression Regulation, Plant
3.Functional mechanisms of WRKY transcription factors in regulating plant response to abiotic stresses.
Shuye WANG ; Guoqiang WU ; Ming WEI
Chinese Journal of Biotechnology 2024;40(1):35-52
WRKYs is a unique family of transcription factors (TFs) in plants, and belongs to the typical multifunctional regulator. It is involved in the regulation of multiple signaling pathways. This type of transcription factor is characterized to contain about 60 highly conservative amino acids as the WRKY domain, and usually also has the Cys2His2 or Cys2His-Cys zinc finger structure. WRKYs can directly bind to the W-box sequence ((T)(T) TGAC (C/T)) in the promoter region of the downstream target gene, and activate or inhibit the transcription of the target genes by interacting with the target protein. They may up-regulate the expression of stress-related genes through integrating signal pathways mediated by abscisic acid (ABA) and reactive oxygen species (ROS), thus playing a vital role in regulating plant response to abiotic stresses. This review summarizes the advances in research on the structure and classification, regulatory approach of WRKYs, and the molecular mechanisms of WRKYs involved in response to drought and salt stresses, and prospects future research directions, with the aim to provide a theoretical support for the genetic improvement of crop in response to abiotic stresses.
Transcription Factors/genetics*
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Abscisic Acid
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Amino Acids
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Droughts
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Stress, Physiological/genetics*
4.Identification of banana ADA1 gene family members and their expression profiles under biotic and abiotic stresses.
Qiqi ZHAO ; Wenhui REN ; Huifei ZHU ; Qiuzhen WU ; Chunyu ZHANG ; Xiaoqiong XU ; Binbin LUO ; Yuji HUANG ; Yukun CHEN ; Yuling LIN ; Zhongxiong LAI
Chinese Journal of Biotechnology 2024;40(1):190-210
The Spt-Ada-Gcn5-acetyltransferase (SAGA) is an ancillary transcription initiation complex which is highly conserved. The ADA1 (alteration/deficiency in activation 1, also called histone H2A functional interactor 1, HFI1) is a subunit in the core module of the SAGA protein complex. ADA1 plays an important role in plant growth and development as well as stress resistance. In this paper, we performed genome-wide identification of banana ADA1 gene family members based on banana genomic data, and analyzed the basic physicochemical properties, evolutionary relationships, selection pressure, promoter cis-acting elements, and its expression profiles under biotic and abiotic stresses. The results showed that there were 10, 6, and 7 family members in Musa acuminata, Musa balbisiana and Musa itinerans. The members were all unstable and hydrophilic proteins, and only contained the conservative SAGA-Tad1 domain. Both MaADA1 and MbADA1 have interactive relationship with Sgf11 (SAGA-associated factor 11) of core module in SAGA. Phylogenetic analysis revealed that banana ADA1 gene family members could be divided into 3 classes. The evolution of ADA1 gene family members was mostly influenced by purifying selection. There were large differences among the gene structure of banana ADA1 gene family members. ADA1 gene family members contained plenty of hormonal elements. MaADA1-1 may play a prominent role in the resistance of banana to cold stress, while MaADA1 may respond to the Panama disease of banana. In conclusion, this study suggested ADA1 gene family members are highly conserved in banana, and may respond to biotic and abiotic stress.
Musa/genetics*
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Phylogeny
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Fungal Proteins
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Cell Nucleus
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Histones
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Stress, Physiological/genetics*
5.Overexpression of Spinacia oleracea betaine aldehyde dehydrogenase (SoBADH) gene confers the salt and cold tolerant in Gossypium hirsutum L.
Xiaoli LUO ; Juanli XIAO ; Zhian WANG ; Anhong ZHANG ; Yingchuan TIAN ; Jiahe WU
Chinese Journal of Biotechnology 2008;24(8):1464-1469
The open reading frame of Spinacia oleracea Betaine Aldehyde Dehydrogenase (SoBADH) was retrieved from Spinacia oleracea and inserted into the Agrobacterium tumefaciens binary vector pBin438, which was driven by CaMV35S promoter, and produced the new binary vector pBSB. A. tumefaciens LBA4404 carrying this plasmid was used in genetic transformation of plants. Forty-five primary transgenic plants were detected by PCR and verified by the Southern blotting from 65 regenerated plants, of which 27 transgenic plants had only one copy of T-DNA. The Northern blotting and Western blotting analysis indicated that the SoBADH gene had been transcribed mRNA and expression protein in the transgenic cotton lines. The testing of SoBADH activity of transgenic plant leaves showed that the enzyme activity was much higher than that of the non-transgenic cotton. The growth of transgenic plants was well under the salinity and freezing stress, whereas the non-transgenic plant grew poorly and even died. Challenging with salinity, the height and fresh weight of transgenic plants was higher compared with those of non-transgenic plants. Under the freezing stress, the relative conductivity of leaf electrolyte leakage of the transgenic cotton lines was lower than that of non-transgenic plants. These results demonstrated that the SoBADH gene could over express in the exogenous plants, and could be used in genetic engineering for cotton stress resistance.
Adaptation, Physiological
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Betaine-Aldehyde Dehydrogenase
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biosynthesis
;
genetics
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Cold Temperature
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Gossypium
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enzymology
;
genetics
;
Plants, Genetically Modified
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enzymology
;
genetics
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Salinity
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Spinacia oleracea
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enzymology
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genetics
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Stress, Physiological
;
genetics
6.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
;
physiology
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Arabidopsis Proteins
;
genetics
;
metabolism
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Autophagy
;
genetics
;
Disease Resistance
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Plant Diseases
;
immunology
;
Saccharomyces cerevisiae
;
genetics
;
Sequence Homology
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Stress, Physiological
7.Chronic Stress Induces Neurotrophin-3 in Rat Submandibular Gland.
Juri SARUTA ; Michitaro IIDA ; Yusuke KONDO ; Masahiro TO ; Takashi HAYASHI ; Mayumi HORI ; Sadao SATO ; Keiichi TSUKINOKI
Yonsei Medical Journal 2012;53(6):1085-1092
PURPOSE: Plasma neurotrophin-3 (NT-3) levels are associated with several neural disorders. We previously reported that neurotrophins were released from salivary glands following acute immobilization stress. While the salivary glands were the source of plasma neurotrophins in that situation, the association between the expression of neurotrophins and the salivary gland under chronic stress conditions is not well understood. In the present study, we investigated whether NT-3 levels in the salivary gland and plasma were influenced by chronic stress. MATERIALS AND METHODS: Expressions of NT-3 mRNA and protein were characterized, using real-time polymerase chain reactions, enzyme-linked immunosorbent assay, and immunohistochemistry, in the submandibular glands of male rats exposed to chronic stress (12 h daily for 22 days). RESULTS: Plasma NT-3 levels were significantly increased by chronic stress (p<0.05), and remained elevated in bilaterally sialoadenectomized rats under the same condition. Since chronic stress increases plasma NT-3 levels in the sialoadenectomized rat model, plasma NT-3 levels were not exclusively dependent on salivary glands. CONCLUSION: While the salivary gland was identified in our previous study as the source of plasma neurotrophins during acute stress, the exposure to long-term stress likely affects a variety of organs capable of releasing NT-3 into the bloodstream. In addition, the elevation of plasma NT-3 levels may play important roles in homeostasis under stress conditions.
Animals
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Male
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Neurotrophin 3/*blood/genetics
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Rats
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Rats, Sprague-Dawley
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Stress, Physiological/*physiology
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Submandibular Gland/*metabolism
8.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
9.Mechanomics: an emerging field between biology and biomechanics.
Jiawen WANG ; Dongyuan LÜ ; Debin MAO ; Mian LONG
Protein & Cell 2014;5(7):518-531
Cells sense various in vivo mechanical stimuli, which initiate downstream signaling to mechanical forces. While a body of evidences is presented on the impact of limited mechanical regulators in past decades, the mechanisms how biomechanical responses globally affect cell function need to be addressed. Complexity and diversity of in vivo mechanical clues present distinct patterns of shear flow, tensile stretch, or mechanical compression with various parametric combination of its magnitude, duration, or frequency. Thus, it is required to understand, from the viewpoint of mechanobiology, what mechanical features of cells are, why mechanical properties are different among distinct cell types, and how forces are transduced to downstream biochemical signals. Meanwhile, those in vitro isolated mechanical stimuli are usually coupled together in vivo, suggesting that the different factors that are in effect individually could be canceled out or orchestrated with each other. Evidently, omics analysis, a powerful tool in the field of system biology, is advantageous to combine with mechanobiology and then to map the full-set of mechanically sensitive proteins and transcripts encoded by its genome. This new emerging field, namely mechanomics, makes it possible to elucidate the global responses under systematically-varied mechanical stimuli. This review discusses the current advances in the related fields of mechanomics and elaborates how cells sense external forces and activate the biological responses.
Biomechanical Phenomena
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Gene Expression Regulation
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Humans
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Mechanotransduction, Cellular
;
Models, Biological
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Proteome
;
genetics
;
metabolism
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Stress, Physiological
;
Transcriptome
10.The responsive characteristics of phytochrome genes to photoperiod, abiotic stresses and identification of their key natural variation sites in foxtail millet (Setaria italica L.).
Xiaoping JIA ; Bo ZHANG ; Zhanxiang HE ; Jianfeng LI ; Xiaomei ZHANG ; Di GE ; Zhenshan WANG ; Luman SANG ; Zhiwei SONG
Chinese Journal of Biotechnology 2022;38(5):1929-1945
The responsive patterns of phytochrome gene family members to photoperiod and abiotic stresses were comparatively analyzed and the favorable natural variation sites of these genes were identified. This would help understand the mechanism of phytochrome gene family in photoperiod-regulated growth and development and abiotic stress response. In addition, it may facilitate the molecular marker assisted selection of key traits in foxtail millet. In this study, we used RT-PCR to clone three phytochrome genes SiPHYA, SiPHYB and SiPHYC from ultra-late maturity millet landrace variety 'Maosu'. After primary bioinformatics analysis, we studied the photoperiod control mode and the characteristics of these genes in responding to five abiotic stresses including polyethylene glycol (PEG)-simulated drought, natural drought, abscisic acid (ABA), high temperature and NaCl by fluorescence quantitative PCR. Finally, we detected the mutation sites of the three genes among 160 foxtail millet materials and performed haplotype analysis to determine the genes' functional effect. We found that the cloned cDNA sequences of gene SiPHYA, SiPHYB and SiPHYC were 3 981, 3 953 and 3 764 bp respectively, which contained complete coding regions. Gene SiPHYB and SiPHYC showed closer evolutionary relationship. Photoperiod regulated all of the three genes, but showed more profound effects on diurnal expression pattern of SiPHYB, SiPHYC than that of SiPHYA. Under short-day, when near heading, the expression levels of SiPHYA and SiPHYB were significantly lower than that under long-day, indicating their roles in suppressing heading of foxtail millet under long-day. SiPHYB and SiPHYC were responsive to PEG-simulated drought, natural drought, ABA and high temperature stresses together. SiPHYA and SiPHYB responded differently to salt stress, whereas SiPHYC did not respond to salt stress. Re-sequencing of 160 foxtail millet materials revealed that SiPHYB was highly conservative. Two missense mutations of SiPHYA, such as single nucleotide polymorphism (SNP) 7 034 522C→T and SNP7 036 657G→C, led to delaying heading and increasing plant height. One missense mutation of SiPHYC, such as SNP5 414 823G→T, led to shortening heading under short-day and delaying heading under long-day, as well as increasing plant height and panicle length regardless of photo-thermal conditions. Photoperiod showed different regulatory effects on SiPHYA, SiPHYB and SiPHYC. SiPHYB and SiPHYC jointly responded to various abiotic stresses except for the salt stress. Compared with the reference genotype, mutation genotypes of SiPHYA and SiPHYC delayed heading and increased plant height and panicle length.
Gene Expression Regulation, Plant
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Photoperiod
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Phytochrome/metabolism*
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Plant Proteins/metabolism*
;
Setaria Plant/metabolism*
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Stress, Physiological/genetics*