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Yeast one-hybrid analyses of the factors binding to W-box elements.A. Scheme of reporter and effector vectors. Fragments of the TaeIF5A1 promoter were inserted into the upstream of the His3 reporter gene. Promoter fragments of 461 bp and 165 bp containing the W-box motif (construct R1, R2) and promoter fragment of 165 bp containing the mutated sequence (construct mR2) were tested. The W-box and mutated W-boxes were respectively cloned into the upstream of the His3 reporter gene: W-box (construct R3); the mutants (constructs R4–R6). The effector vectors, E1: pGADT7-Rec2 harboring TaRAV; E2: pGADT7-Rec2 harboring TaWRKY. B. TaRAV and TaWRKY interaction with W-box (R3) or mutants (R4–R6) sequences. C. Determination of TaRAV and TaWRKY binding to the promoter fragments containing W-box or mutated motif (R1, R2, mR2). The effector and the reporter constructs were co-transformed into yeast strain Y187. Positive transformants were determined by spotting serial dilutions (1:1, 1:10, 1:100, 1:1000) of yeast onto SD/-His/-Leu/-Trp plates with 3-AT. Negative controls: N1, p53HIS2 + E1 (AD-TaRAV); N2, p53HIS2 + E2 (AD-TaWRKY); Positive control: P, p53HIS2 + pGAD-Rec2-53. D. TaRAV and TaWRKY binding to the W-box and the promoter fragment containing the W-box motif in tobacco leaves. (a) Construction of reporter and effector plasmids for transient trans-activation assays; (b) GUS staining of tobacco leaves co-transformed with reporter and effector plasmids; (c) GUS activity assay of the co-expression of effector and reporter plasmids. The data represent mean values of three independent experiments.
Source publication
The eukaryotic translation initiation factor 5A (eIF5A) promotes formation of the first peptide bond at the onset of protein synthesis. However, the function of eIF5A in plants is not well understood.
In this study, we characterized the function of eIF5A (TaeIF5A1) from Tamarix androssowii. The promoter of TaeIF5A1 with 1,486 bp in length was isola...
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Sucrose non-fermenting 1-related protein kinase 2 (SnRK2) family members play crucial roles in plant abiotic stress response. However, the precise mechanism underlying the function of SnRKs has not been thoroughly elucidated in plants. In this research, a novel SnRK2 gene, TaSnRK2.9 was cloned and characterized from common wheat. The expression of...
Citations
... After a treatment duration of 10 days, we assessed the activity of superoxide dismutase (SOD) and the levels of malondialdehyde (MDA) and proline in both the transgenic Arabidopsis and wild-type specimens. These assessments were carried out following established protocols [24,25]. Each sample consisted of three distinct plants, and experiments related to physiological indices were performed in triplicate. ...
Aquaporins play a crucial role in helping water molecules move across cell membranes. While some studies have examined the role of AQPs in model plants like Arabidopsis, their impact on the ability of non-model plants to withstand environmental stress is largely unknown. In this study, we have explored the functions of the PaPIP1-2 gene, which encodes a protein called PIP, in apricot kernels (Prunus armeniaca L.). Our findings reveal how the PaPIP1-2 gene behaves during both dormancy and sprouting phases. Using a network analysis, we identified its interaction with six genes related to cold resistance. The mRNA levels of PaAQP genes, which co-express with cold resistance genes, remain consistent throughout different stages of P. armeniaca flower bud development, including physiological dormancy (PD), ecological dormancy (ED), sprouting period (SP), and germination stage (GS). Furthermore, our investigation of the location of the GFP-tagged PaPIP1-2 protein showed that it is mainly found in the cell membrane. Yeast strains with overexpressed PaPIP1-2 exhibited improved cold resistance and higher protein content. Similarly, when we overexpressed PaPIP1-2 in Arabidopsis, it enhanced the growth of these transgenic plants under cold stress. This improvement was associated with reduced levels of MDA (malondialdehyde); decreased ion leakage; increased proline accumulation; superoxide dismutase (SOD) activity; and the expression of cold resistance genes like AtPUB26, AtBTF3L, AtEBF1-1, and AtRAV1, compared with the wild-type plants. In summary, our results highlight the role of the P. armeniaca PaPIP1-2 gene in enhancing cold resistance and its importance in the dormancy and germination stages.
... On the basis of identified interacting partners, a hypothetical model has been suggested (Fig. 6), which highlights importance of OsRuvBL1a in providing stress tolerance. ROS production and lipid peroxidation are major products of stresses (Sewelam et al. 2016) and OsRuvBL1a controls ROS scavenging pathway genes through hormonal regulation by interacting with proteins such as SPINDLY (Furstenberg-Hagg et al. 2013;Shimada et al. 2006), START (Hubbard et al. 2010;Klingler et al. 2010), eIF5a (Wang et al. 2012) OsIAA3 and ZIM domain containing proteins (Ishiga et al. 2013;Wang et al. 2017). ...
RuvBL, is a member of SF6 superfamily of helicases and is conserved among the various model systems. Recently, rice (Oryza sativa L.) homolog of RuvBL has been biochemically characterized for its ATPase and DNA helicase activities; however its involvement in stress has not been studied so far. Present investigation reports the detailed functional characterization of OsRuvBL under abiotic stresses through genetic engineering. An efficient Agrobacterium-mediated in planta transformation protocol was developed in indica rice to generate the transgenic lines and study was focused on optimization of factors to achieve maximum transformation efficiency. Overexpressing OsRuvBL1a transgenic lines showed enhanced tolerance under in vivo salinity stress as compared to WT plants. The physiological and biochemical analysis of the OsRuvBL1a transgenic lines showed better performance under salinity and drought stresses. Several stress responsive interacting partners of OsRuvBL1a were identified using Y2H system revealed to its role in stress tolerance. Functional mechanism for boosting stress tolerance by OsRuvBL1a has been proposed in this study. This integration of OsRuvBL1a gene in rice genome using in planta transformation method helped to achieve the abiotic stress resilient smart crop. This study is the first direct evidence to show the novel function of RuvBL in boosting abiotic stress tolerance in plants.
... O 2 and H 2 O 2 contents were detected using the methods in our previous work [35]. SOD, POD, and CAT activities were measured using the method of Wang et al. [36]. MDA content and electrolyte leakage were detected according to Fu et al. [37]. ...
Low temperatures restrict the growth of the grapevine industry. The DREB transcription factors are involved in the abiotic stress response. Here, we isolated the VvDREB2A gene from Vitis vinifera cultivar ‘Zuoyouhong’ tissue culture seedlings. The full-length VvDREB2A cDNA was 1068 bp, encoding 355 amino acids, which contained an AP2 conserved domain belonging to the AP2 family. Using transient expression in leaves of tobacco, VvDREB2A was localized to the nucleus, and it potentiated transcriptional activity in yeasts. Expression analysis revealed that VvDREB2A was expressed in various grapevine tissues, with the highest expression in leaves. VvDREB2A was induced by cold and the stress-signaling molecules H2S, nitric oxide, and abscisic acid. Furthermore, VvDREB2A-overexpressing Arabidopsis was generated to analyze its function. Under cold stress, the Arabidopsis overexpressing lines exhibited better growth and higher survival rates than the wild type. The content of oxygen free radicals, hydrogen peroxide, and malondialdehyde decreased, and antioxidant enzyme activities were enhanced. The content of raffinose family oligosaccharides (RFO) also increased in the VvDREB2A-overexpressing lines. Moreover, the expression of cold stress-related genes (COR15A, COR27, COR6.6, and RD29A) was also enhanced. Taken together, as a transcription factor, VvDREB2A improves plants resistance to cold stress by scavenging reactive oxygen species, increasing the RFO amount, and inducing cold stress-related gene expression levels.
... Li et al. (2015) also reported higher abundance of elongation factor gi|6015064 under salt stress conditions in cotton. Among different families of eukaryotic initiation factors (eIFs), the role of eIF1A has been demonstrated in abiotic stress tolerance (Singh et al., 2007;Wang et al., 2012). Li et al. (2019) identified 18 eIF genes from mango transcriptome and found that majority of the MieIF genes were involved with abiotic stress in mango among which the expression ofMieIF1A-a, significantly increased under salinity stress.The upregulation various translation related transcripts in Turpentine indicates the involvement of synthesis of stress related proteins to help it cope with salinity stress. ...
Introduction
Increased soil salinity in the recent years has adversely affected the productivity of mango globally. Extending the cultivation of mango in salt affected regions warrants the use of salinity tolerant/resistant rootstocks. However, the lack of sufficient genomic and transcriptomic information impedes comprehensive research at the molecular level.
Method
We employed RNA sequencing-based transcriptome analysis to gain insight into molecular response to salt stress by using two polyembryonic mango genotypes with contrasting response to salt stress viz., salt tolerant Turpentine and salt susceptible Mylepelian.
Results
RNA sequencing by Novaseq6000 resulted in a total of 2795088, 17535948, 7813704 and 5544894 clean reads in Mylepelian treated (MT), Mylepelian control (MC), Turpentine treated (TT) and Turpentine control (TC) respectively. In total, 7169 unigenes annotated against all the five public databases, including NR, NT, PFAM, KOG, Swissport, KEGG and GO. Further, maximum number of differentially expressed genes were found between MT and MC (2106) followed by MT vs TT (1158) and TT and TC (587). The differentially expressed genes under different treatment levels included transcription factors (bZIP, NAC, bHLH), genes involved in Calcium-dependent protein kinases (CDPKs), ABA biosynthesis, Photosynthesis etc. Expression of few of these genes was experimentally validated through quantitative real-time PCR (qRT-PCR) and contrasting expression pattern of Auxin Response Factor 2 (ARF2), Late Embryogenesis Abundant (LEA) and CDPK genes were observed between Turpentine and Mylepelian.
Discussion
The results of this study will be useful in understanding the molecular mechanism underlying salt tolerance in mango which can serve as valuable baseline information to generate new targets in mango breeding for salt tolerance.
... Among these upregulated proteins, the function of TIF4A may be to increase the accumulation of superoxide dismutase, scavenge the reactive oxygen species, and enhance membrane stability [28]. In addition, high accumulation of TIF4A induces cellular reorganization as well as prevents chlorophyll loss and membrane damage under stress exposure [29,30]. Therefore, TIF4A may play an important role in the adaptation of C. lentillifera to changes in environmental nitrate concentrations. ...
... Evans blue, diaminobenzidine (DAB), and nitroblue tetrazolium (NBT) staining methods were performed according to Gong et al. (2005) and Qu et al. (2013). The activities of peroxidase (POD) and superoxide dismutase (SOD) were measured according to the methods of Wang et al. (2012). Proline content was determined using a proline assay kit (Nanjing Jiancheng Bioengineering Institute, China), according to the manufacturer's protocol. ...
Key message
The drought response lncRNAs were identified from birch, and the lncRNAs involved in drought tolerance were characterized.
Abstract
Long non-coding RNAs (lncRNAs) participate in many biological processes. However, there is limited information on the role of lncRNAs in the abiotic stress tolerance of trees. Here, we identified lncRNAs from the birch Betula platyphylla and characterized their functions in drought tolerance. A total of 3670 lncRNAs were identified, which were predicted to regulate 15843 target genes by trans-regulation and/or cis-regulation. A total of 53 drought-responsive lncRNAs were identified. Six drought-responsive lncRNAs were randomly selected for further investigation. The predicted target genes were verified using qRT-PCR to compare their expressions in plants that transiently expressed the lncRNAs and control. These lncRNAs were transiently transformed into birch for overexpression and RNAi silencing, and the physiological and biochemical parameters related to drought tolerance were measured. Of the six studied lncRNAs, four lncRNAs could confer drought tolerance to the transgenic plants, and expression of the other two lncRNAs showed sensitivity to drought stress. These results suggested that many birch drought responsive lncRNAs were involved in drought tolerance; thereby, characterization of their function is necessary for revealing the drought tolerance mechanism of birch.
... Similarly, we also identified eukaryotic translation initiation factor 5A (eIF5A), which is upregulated in low and high doses of UV-B, whereas it was downregulated in the middle dose of UV-B. TaeIF5A1 from Tamarix androssowii, TaWRKY, and RAV protein share very similar expression patterns, and all stressresponsive genes are involved in the ABA signaling pathway (Wang et al., 2012). eIF5A-2 from Arabidopsis played a crucial role in plant growth and development by regulating cell division, cell growth, and cell death (Feng et al., 2007). ...
Rice (Oryza sativa) is a human staple food and serves as a model organism for genetic and molecular studies. Few studies have been conducted to determine the effects of ultraviolet-B (UV-B) stress on rice. UV-B stress triggers morphological and physiological changes in plants. However, the underlying mechanisms governing these integrated responses are unknown. In this study, we conducted a proteomic response of rice leaves to UV-B stress using two-dimensional gel electrophoresis and identified the selected proteins by mass spectrometry analysis. Four levels of daily biologically effective UV-B radiation intensities were imposed to determine changes in protein accumulation in response to UV-B stress: 0 (control), 5, 10, and 15 kJ m⁻² d⁻¹in two cultivars, i.e., IR6 and REX. To mimic the natural environment, we conducted this experiment in Sunlit Soil-Plant-Atmosphere-Research (SPAR) chambers. Among the identified proteins, 11% of differentially expressed proteins were found in both cultivars. In the Rex cultivar, only 45% of proteins are differentially expressed, while only 27.5% were expressed in IR6. The results indicate that REX is more affected by UV-B stress than IR6 cultivars. The identified protein TSJT1 (spot 16) in both cultivars plays a crucial role in plant growth and development during stress treatment. Additionally, we found that UV-B stress altered many antioxidant enzymes associated with redox homeostasis and cell defense response. Another enzyme, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), has been identified as spot 15, which plays an essential role in glycolysis and cellular energy production. Another vital protein identified is glycosyl hydrolase (GH) as spot 9, which catalyzes the hydrolysis of glycosidic bonds in cell wall polymers and significantly affects cell wall architecture. Some identified proteins are related to photosynthesis, protein biosynthesis, signal transduction, and stress response. The findings of our study provide new insights into understanding how rice plants are tailored to UV-B stress via modulating the expression of UV-B responsive proteins, which will help develop superior rice breeds in the future to combat UV-B stress. Data are available via ProteomeXchange with identifier PXD032163.
... There are indications that eIF5A can play a particular role for stress tolerance. Ectopic overexpression of yeast eIF5A yielded transgenic yeast or poplar plants with increased resistance to oxidative and hyperosmotic stress (Wang et al. 2012). It has also been observed that eIF5A deficiency causes increased sensitivity to acetic acid stress in yeast (Cheng et al. 2021). ...
To remain competitive, cells exposed to stress of varying duration, rapidity of onset, and intensity, have to balance their expenditure on growth and proliferation versus stress protection. To a large degree dependent on the time scale of stress exposure, the different levels of gene expression control: transcriptional, post-transcriptional and post-translational, will be engaged in stress responses. The post-transcriptional level is appropriate for minute-scale responses to transient stress, and for recovery upon return to normal conditions. The turnover rate, translational activity, covalent modifications, and subcellular localisation of RNA species are regulated under stress by multiple cellular pathways. The interplay between these pathways is required to achieve the appropriate signalling intensity and prevent undue triggering of stress-activated pathways at low stress levels, avoid overshoot, and down-regulate the response in a timely fashion.
As much of our understanding of post-transcriptional regulation has been gained in yeast, this review is written with a yeast bias, but attempts to generalise to other eukaryotes. It summarises aspects of how post-transcriptional events in eukaryotes mitigate short-term environmental stresses, and how different pathways interact to optimise the stress response under shifting external conditions.
... The seeds were sown and cultured in Petri dishes containing double-distilled water. When two cotyledons appeared, seedlings with similar height were transferred to woody plant medium (WPM) [29] agar plates supplemented with 0.4 M sodium chloride (NaCl) [30,31] for 0 (control), 2, 4, 6, and 24 h maintained at 25 ± 2 • C with a 16 h light/8 h dark cycle and a light intensity of 1000-1500 lx. All tissues of the seedlings were harvested. ...
... To evaluate the activity of the GUS reporter gene in pBpTCP3::GUS transgenic B. platyphylla under salt treatment, the tissue-cultured rooting seedlings of the pBpTCP3::GUS transgenic line were spread on a medium containing 0.4 M NaCl [30,31] and treated for 0 (control), 2, 4, 6, and 24 h. The seedlings were incubated for GUS staining, which was performed according to the protocol described by Jefferson [37]. ...
The plant-specific transcription factors TEOSINTE BRANCHED1/CYCLO IDEA/PROLIFERATING CELL FACTOR1 (TCP) act as developmental regulators that have many roles in the growth and development processes throughout the entire life span of plants. TCP transcription factors are responsive to endogenous and environmental signals, such as salt stress. However, studies on the role of the TCP genes in salt stress response have rarely focused on woody plants, especially forest trees. In this study, the BpTCP3 gene, a CYC/TB1 subfamily member, isolated from Betula platyphylla Sukaczev, was significantly influenced by salt stress. The β-glucuronidase (GUS) staining analysis of transgenic B. platyphylla harboring the BpTCP3 promoter fused to the reporter gene GUS (pBpTCP3::GUS) further confirmed that the BpTCP3 gene acts a positive regulatory position in salt stress. Under salt stress, we found that the BpTCP3 overexpressed lines had increased relative/absolute high growth but decreased salt damage index, hydrogen peroxide (H2O2), and malondialdehyde (MDA) levels versus wild-type (WT) plants. Conversely, the BpTCP3 suppressed lines exhibited sensitivity to salt stress. These results indicate that the BpTCP3 transcription factor improves the salt tolerance of B. platyphylla by reducing reactive oxygen species damage, which provides useful clues for the functions of the CYC/TB1 subfamily gene in the salt stress response of B. platyphylla.
... After 10 days of treatment, the physiological indexes of transgenic Arabidopsis and wild type plants were determined. Superoxide dismutase (SOD) activity (Wang et al., 2012a) and malondialdehyde (MDA) and proline levels were measured as previously described (Bates et al., 1973). Each sample comprised at least three individual plants, and all physiological analysis experiments were conducted in triplicate. ...
Aquaporins (AQPs) are essential channel proteins that play a major role in plant growth and development, regulate plant water homeostasis, and transport uncharged solutes across biological membranes. In this study, 33 AQP genes were systematically identified from the kernel-using apricot (Prunus armeniaca L.) genome and divided into five subfamilies based on phylogenetic analyses. A total of 14 collinear blocks containing AQP genes between P. armeniaca and Arabidopsis thaliana were identified by synteny analysis, and 30 collinear blocks were identified between P. armeniaca and P. persica. Gene structure and conserved functional motif analyses indicated that the PaAQPs exhibit a conserved exon-intron pattern and that conserved motifs are present within members of each subfamily. Physiological mechanism prediction based on the aromatic/arginine selectivity filter, Froger’s positions, and three-dimensional (3D) protein model construction revealed marked differences in substrate specificity between the members of the five subfamilies of PaAQPs. Promoter analysis of the PaAQP genes for conserved regulatory elements suggested a greater abundance of cis-elements involved in light, hormone, and stress responses, which may reflect the differences in expression patterns of PaAQPs and their various functions associated with plant development and abiotic stress responses. Gene expression patterns of PaAQPs showed that PaPIP1-3, PaPIP2-1, and PaTIP1-1 were highly expressed in flower buds during the dormancy and sprouting stages of P. armeniaca. A PaAQP coexpression network showed that PaAQPs were coexpressed with 14 cold resistance genes and with 16 cold stress-associated genes. The expression pattern of 70% of the PaAQPs coexpressed with cold stress resistance genes was consistent with the four periods [Physiological dormancy (PD), ecological dormancy (ED), sprouting period (SP), and germination stage (GS)] of flower buds of P. armeniaca. Detection of the transient expression of GFP-tagged PaPIP1-1, PaPIP2-3, PaSIP1-3, PaXIP1-2, PaNIP6-1, and PaTIP1-1 revealed that the fusion proteins localized to the plasma membrane. Predictions of an A. thaliana ortholog-based protein–protein interaction network indicated that PaAQP proteins had complex relationships with the cold tolerance pathway, PaNIP6-1 could interact with WRKY6, PaTIP1-1 could interact with TSPO, and PaPIP2-1 could interact with ATHATPLC1G. Interestingly, overexpression of PaPIP1-3 and PaTIP1-1 increased the cold tolerance of and protein accumulation in yeast. Compared with wild-type plants, PaPIP1-3- and PaTIP1-1-overexpressing (OE) Arabidopsis plants exhibited greater tolerance to cold stress, as evidenced by better growth and greater antioxidative enzyme activities. Overall, our study provides insights into the interaction networks, expression patterns, and functional analysis of PaAQP genes in P. armeniaca L. and contributes to the further functional characterization of PaAQPs.
