SlMYB17 Antagonises the SlCBF Pathway to Negatively Regulate Tomato Chilling Tolerance.

AP2/ERF family transcription factors in plant abiotic stress responses

GIGANTEA functions as a co-repressor of cold stress response with a histone-modifying complex.

Like many plants, Arabidopsis thaliana increases in freezing tolerance in response to low non-freezing temperatures, a phenomenon known as cold acclimation. Associated with cold acclimation are a number of biochemical changes including the expression of COR (cold-regulated) genes. Here we summarize recent progress we have made in understanding the function and regulation of these genes. One significant finding regarding COR gene function is that constitutive expression of COR15a in transgenic Arabidopsis plants enhances the freezing tolerance of both chloroplasts and protoplasts. These results provide the first direct evidence for a COR gene having a role in freezing tolerance. The precise mechanism of COR15a action is not yet know, but current results indicate the gene has a role in stabilizing membranes against freeze-induced damage. In regards to COR gene regulation, we have isolated a cDNA for CBF1, the first identified transcriptional activator that binds to the CRT (C-repeat)/DRE (drought responsive element), a cold- and drought-responsive DNA regulatory element present in the promoters of COR genes. Our working hypothesis is that CBF1 binds to the CRT/DRE sequence and participates in the regulation of COR genes in response to low temperature and drought.

Understanding the principles of abiotic and biotic stress responses, tolerance and adaptation remains important in plant physiology research to develop better varieties of crop plants. Better understanding of plant stress response mechanisms and application of knowledge derived from integrated experimental and bioinformatics approaches are gaining importance. Earlier, we showed that compiling a database of stress-responsive transcription factors and their corresponding target binding sites in the form of Hidden Markov models at promoter, untranslated and upstream regions of stress-up-regulated genes from expression analysis can help in elucidating various aspects of the stress response in Arabidopsis. In addition to the extensive content in the first version, STIFDB2 is now updated with 15 stress signals, 31 transcription factors and 5,984 stress-responsive genes from three species (Arabidopsis thaliana, Oryza sativa subsp. japonica and Oryza sativa subsp. indica). We have employed an integrated biocuration and genomic data mining approach to characterize the data set of transcription factors and consensus binding sites from literature mining and stress-responsive genes from the Gene Expression Omnibus. STIFDB2 currently has 38,798 associations of stress signals, stress-responsive genes and transcription factor binding sites predicted using the Stress-responsive Transcription Factor (STIF) algorithm, along with various functional annotation data. As a unique plant stress regulatory genomics data platform, STIFDB2 can be utilized for targeted as well as high-throughput experimental and computational studies to unravel principles of the stress regulome in dicots and gramineae. STIFDB2 is available from the URL: http://caps.ncbs.res.in/stifdb2

Involvement of plant C2H2-type zinc finger transcription factors in stress responses

SummaryA model is presented describing the gene regulatory network surrounding three similar NAC transcription factors that have roles in Arabidopsis leaf senescence and stress responses. ANAC019, ANAC055 and ANAC072 belong to the same clade of NAC domain genes and have overlapping expression patterns. A combination of promoter DNA/protein interactions identified using yeast 1-hybrid analysis and modelling using gene expression time course data has been applied to predict the regulatory network upstream of these genes. Similarities and divergence in regulation during a variety of stress responses are predicted by different combinations of upstream transcription factors binding and also by the modelling. Mutant analysis with potential upstream genes was used to test and confirm some of the predicted interactions. Gene expression analysis in mutants of ANAC019 and ANAC055 at different times during leaf senescence has revealed a distinctly different role for each of these genes. Yeast 1-hybrid analysis is shown to be a valuable tool that can distinguish clades of binding proteins and be used to test and quantify protein binding to predicted promoter motifs.

Abiotic stress factors adversely affect crop growth and yield all over the world. NAC transcription factors could regulate stress response in plant species, and their underlying mechanisms should be studied. The current work investigates whether the rice transcription factor, SNAC3, could augment salinity tolerance of rice plants. Results indicated that salt-stressed rice plants overexpressing SNAC3 had improved salt tolerance and yield, enhanced relative water content and osmolytes, increased gas-exchange attributes and antioxidant enzymes activity, up-regulated stress-responsive genes expression, and reduced oxidative stress markers levels, as compared to wild-type plants. SNAC3 rice mutants exhibited the reversed traits. Additionally, simple sequence repeats analysis showed genetically diverse patterns among treated and non-treated plants. In conclusion, SNAC3 has a crucial role in improving salinity tolerance and grain yield of rice plants. This study also presented transgenic rice lines with enhanced grain yield and improved salinity tolerance.

NAC proteins are plant-specific transcription factors that play essential roles in stress responses. However, only little information regarding stress-related NAC genes is available in maize. In this study, a maize NAC gene, ZmSNAC1, was cloned and functionally characterized. Expression analysis revealed that ZmSNAC1 was strongly induced by low temperature, high-salinity, drought stress, and abscisic acid (ABA) treatment, but downregulated by salicylic acid treatment. Subcellular localization experiments in Arabidopsis protoplast cells indicated that ZmSNAC1 was localized in the nucleus. Transactivation assays demonstrated that ZmSNAC1 functioned as a transcriptional activator. Overexpression of ZmSNAC1 in Arabidopsis led to hypersensitivity to ABA and osmotic stress at the germination stage, but enhanced tolerance to dehydration compared to wild-type seedlings. These results suggest that ZmSNAC1 functions as a stress-responsive transcription factor in positive modulation of abiotic stress tolerance, and may have applications in the engineering of drought-tolerant crops. ZmSNAC1 functioned as a stress-responsive transcription factor in response to abiotic stresses, and might be useful for crop tolerance improvement.

SlNAC4 functions as a stress-responsive transcription factor and might be useful for crop salt and drought tolerance improvement. Abiotic stresses, especially salinity and drought, are major factors that significantly limit crop growth and productivity. Plant-specific NAC transcription factors play crucial roles in various stress responses. However, to date only little information regarding stress-related NAC genes is available in tomato. Previously, we reported that tomato SlNAC4-SlNAC10 genes are involved in response of various abiotic stresses. Expression analysis revealed that SlNAC4 was also induced significantly by MeJA, but not by ABA. To further unravel the function of SlNAC4 in response to abiotic stress, we investigated the effects of salt and drought stress on wild-type and SlNAC4-RNAi transgenic tomato plants. The results demonstrated that the root and shoot growth of RNAi plants was more inhibited by salt stress than that of wild-type at post-germination stage. The leaf salt assay also showed less tolerance in transgenic plants by retaining lower chlorophyll content compared with wild-type plants. In addition, transgenic plants became less tolerant to salt and drought stress in soil, which were demonstrated by lower levels of water and chlorophyll contents, and higher water loss rate in their leaves as compared to wild-type plants under stressed conditions. Notably, the expressions of multiple stress-related genes were downregulated in SlNAC4-RNAi plants under both control and salt-stressed conditions. Collectively, these results highlight the important role of SlNAC4 functions as a stress-responsive transcription factor in positive modulation of abiotic stress tolerance through an ABA-independent signaling networks and possibly in response to biotic stress, and may hold promising applications in the engineering of salt- and drought-tolerant tomato.

Pre-implantation embryo development is a critical stage, in which several development and stress-response related transcription factors (TF) are involved. Exposing embryos to environmental insults alter some of these stress response-related TF. However, their expression pattern in male and female embryos and their release via exosomes is still unclear. Here, we aimed to investigate the effect of culture-induced oxidative stress on development and expression pattern of stress-related TF in male and female embryos and in respective spent media coupled with exosomes. For this, bovine male and female zygotes were in vitro produced using sexed semen and cultured under 5% and 20% oxygen in exosome-depleted SOFaa medium (SOF with amino acids). Blastocysts were subjected to total RNA isolation followed by quantitative RT-PCR analysis of the selected TF (Nrf2, KLF4, NOTCH1, SREBF2, E2F1, CAT1, SOD1, and OCT4), as well as protein abundance analysis using immunofluorescence and related phenotypes analysis, including reactive oxygen species (ROS) level and total cell count. Furthermore, the spent embryo culture media were collected for exosomes isolation and expression analysis of candidate TF. The data were statistically analysed using one-way ANOVA followed by multiple pair-wise comparisons using the Tukey post hoc test. Results showed that the blastocyst rates of both male (29.9% v. 34.9%) and female (16.7% v. 26.5%) bovine embryos were significantly lower in 20% than in 5% oxygen level. Female blastocysts subjected to the higher oxygen level showed increased ROS level (37.66±1.70v. 45.32±2.05 in male and 29.42±1.44v. 45.51±2.06 in female) and significantly reduced total cell count compared with the male embryo counterpart (136.55±7.8v. 112.75±2.9 in male and 138.75±2.0v. 88.25±4.3 in female cultured in 5% and 20% oxygen levels, respectively). Consequently, the expression levels of Nrf2, KLF4, SREBF2, CAT1, SOD1, and OCT4 were significantly increased in male embryos exposed to oxidative stress compared with those cultured under the lower oxygen level. However, NOTCH1 and E2F1 were significantly increased in female embryos exposed to oxidative stress compared with the male counterparts. The mRNA level of SREBF2 was significantly increased in male embryos cultured under both 5% and 20% O2 compared with female embryos. The protein expression level of Nrf2 and KLF4 was higher in embryos cultured at 20% v. 5% O2 with greater Nrf2 abundance in male embryos. Consequently, the male embryos produced at 20% O2 released a higher number of exosomes enriched with Nrf2, SOD1, and NOTCH1 mRNA than the other groups. Interestingly, the exosomal mRNA expression level of E2F1 tended to be higher in female embryos exposed to oxidative stress than their male counterparts. Taken together, the male embryos were more tolerant to oxidative stress than female embryos via the activation Nrf2-mediated oxidative stress response and development related TF. The release of these TF via exosomes could enhance cellular homeostasis maintenance under oxidative stress.

Expression analyses indicate the involvement of sunflower WRKY transcription factors in stress responses, and phylogenetic reconstructions reveal the existence of a novel clade in the Asteraceae

DNA-binding with One Finger (Dof) transcription factors are plant-specific regulators involved in diverse biological processes, including growth, development, and stress responses. However, the Dof family in tobacco (Nicotiana tabacum L.) remains poorly characterized. Here, we performed a genome-wide identification and comprehensive analysis of Dof genes in tobacco, revealing 75 NtDof members. Through phylogenetic reconstruction, gene structure analysis, and cis-regulatory element prediction, we classified these genes into seven subfamilies and found that the CDF clade exhibits conserved exon–intron structures, unique motif compositions, and enrichment of stress- and hormone-responsive cis-elements. Expression profiling showed that most NtCDF genes respond dynamically to cold and magnesium deficiency stresses. Virus-induced gene silencing (VIGS) of two representative NtCDF genes, NtDof40 and NtDof63, revealed their opposing roles in cold tolerance: NtDof40 acts as a positive regulator, while NtDof63 functions as a negative regulator. Our findings provide a comprehensive evolutionary framework for the Dof family in tobacco and highlight the functional divergence of CDF subfamily members in abiotic stress adaptation, offering potential targets for stress-resistance breeding.

Cold stress is one of the major abiotic stresses that limit crop production. The ICE-CBF-COR pathway is associated with cold stress response in a wide variety of crop species. However, the ICE-CBF-COR genes has not been well characterized in wheat (Triticum aestivum). This study identified, characterized and examined the expression profiles of the ICE, CBF and COR genes for cold defense in wheat. Five ICE (inducer of CBF expression) genes, 37 CBF (C-repeat binding factor) genes and 11 COR (cold-responsive or cold-regulated) genes were discovered in the wheat genome database. Phylogenetic trees based on all 53 genes revealed that CBF genes were more diverse than ICE and COR genes. Twenty-two of the 53 genes appeared to include 11 duplicated pairs. Twenty rice (Oryza sativa) genes and 21 sorghum (Sorghum bicolor) and maize (Zea mays) genes showed collinearity with the wheat ICE, CBF and COR genes. Transcriptome data and qRT-PCR analyses revealed tissue-specific expression patterns of the ICE, CBF and COR genes, and identified similarities in the expression pattern of genes from the same family when subjected to drought, heat, drought plus heat, and cold stress. These results provide information for better understanding the biological roles of ICE, CBF, COR genes in wheat.

Dynamic environmental changes such as extreme temperature, water scarcity and high salinity affect plant growth, survival, and reproduction. Plants have evolved sophisticated regulatory mechanisms to adapt to these unfavorable conditions, many of which interface with plant hormone signaling pathways. Abiotic stresses alter the production and distribution of phytohormones that in turn mediate stress responses at least in part through hormone- and stress-responsive transcription factors. Among these, the APETALA2/ETHYLENE RESPONSIVE FACTOR (AP2/ERF) family transcription factors (AP2/ERFs) have emerged as key regulators of various stress responses, in which they also respond to hormones with improved plant survival during stress conditions. Apart from participation in specific stresses, AP2/ERFs are involved in a wide range of stress tolerance, enabling them to form an interconnected stress regulatory network. Additionally, many AP2/ERFs respond to the plant hormones abscisic acid (ABA) and ethylene (ET) to help activate ABA and ET dependent and independent stress-responsive genes. While some AP2/ERFs are implicated in growth and developmental processes mediated by gibberellins (GAs), cytokinins (CTK), and brassinosteroids (BRs). The involvement of AP2/ERFs in hormone signaling adds the complexity of stress regulatory network. In this review, we summarize recent studies on AP2/ERF transcription factors in hormonal and abiotic stress responses with an emphasis on selected family members in Arabidopsis. In addition, we leverage publically available Arabidopsis gene networks and transcriptome data to investigate AP2/ERF regulatory networks, providing context and important clues about the roles of diverse AP2/ERFs in controlling hormone and stress responses.

Increasing evidence has demonstrated that basic leucine zipper (bZIP) transcription factors (TFs) are functional and involved in plant stress response, but few bZIP TFs have thus far been investigated in economically important fruit crops, such as bananas. In our present work, a novel 43.4 kDa bZIP transcription factor, MabZIP3, with a conserved bZIP domain, and a theoretical isoelectric point of 9.96 was isolated and characterized from banana fruit. Phylogenetic analysis showed that MabZIP3 belonged to the AREB subfamily of the bZIP family and was most closely related to BnABF2 and PsABF2. Transient expression analysis of the MabZIP3–GFP fusion protein in tobacco BY2 protoplasts revealed that the MabZIP3 protein is localized in the nucleus. Real-time PCR and promoter analysis of MabZIP3 indicated that it was responsive to methyl jasmonate, abscisic acid, chilling stress and pathogen Colletotrichum musae infection. More importantly, to further understand the regulatory network of MabZIP3 in response to stress, proteins physically interacting with MabZIP3 were identified by screening a cDNA expression library of banana fruit using the yeast two-hybrid system. Several interacting proteins related to stress responses were obtained, including a Saposin-Bdomain-containing protein, MaSAP, and their interaction was further confirmed by bimolecular fluorescence complementation assay (BiFC). Taken together, our results suggest that MabZIP3 is a stress-responsive transcription factor, which might be functionally linked to stress related proteins.