Regulation Of Heat Stress Response Research Articles

Heat shock factor binding protein BrHSBP1 regulates seed and pod development in Brassica rapa.

Plant heat shock factor binding proteins (HSBPs) are well known for their implication in the negative regulation of heat stress response (HSR) pathways. Herein, we report on the hitherto unknown functions of HSBP1 in Brassica rapa (BrHSBP1). BrHBSP1 was found to be predominant in flower buds and young leaves, while its segmental duplicate, BrHSBP1-like, was abundant in green siliques. Exposure to abiotic stress conditions, such as heat, drought, cold, and H2O2, and to phytohormones was found to differentially regulate BrHSBP1. The activity of BrHSBP1-GFP fusion proteins revealed their cellular localization in nuclei and cytosols. Transgenic overexpression of BrHSBP1 (BrHSBP1OX) improved pod and seed sizes, while CRISPR-Cas BrHSBP1 knock-out mutants (Brhsbp1_KO) were associated with aborted seed and pod development. The transcriptomic signatures of BrHSBP1OX and Brhsbp1_KO lines revealed that 360 and 2381 genes, respectively, were differentially expressed (Log2FC≥2, padj<0.05) expressed relative to control lines. In particular, developmental processes, including plant reproductive structure development (RSD)-related genes, were relatively downregulated in Brhsbp1_KO. Furthermore, yeast two-hybrid assays confirmed that BrHSBP1 can physically bind to RSD and other genes. Taking the findings together, it is clear that BrHSBP1 is involved in seed development via the modulation of RSD genes. Our findings represent the addition of a new regulatory player in seed and pod development in B. rapa.

Differential requirement of MED14/17 recruitment for activation of heat inducible genes.

The mechanism of heat stress response in plants has been studied, focusing on the function of transcription factors (TFs). Generally, TFs recruit coactivators, such as Mediator, are needed to assemble the transcriptional machinery. However, despite the close relationship with TFs, how coactivators are involved in transcriptional regulation under heat stress conditions is largely unclear. We found a severe thermosensitive phenotype of Arabidopsis mutants of MED14 and MED17. Transcriptomic analysis revealed that a quarter of the heat stress (HS)-inducible genes were commonly downregulated in these mutants. Furthermore, chromatin immunoprecipitation assay showed that the recruitment of Mediator by HsfA1s, the master regulators of heat stress response, is an important step for the expression of HS-inducible genes. There was a differential requirement of Mediator among genes; TF genes have a high requirement whereas heat shock proteins (HSPs) have a low requirement. Furthermore, artificial activation of HsfA1d mimicking perturbation of protein homeostasis induced HSP gene expression without MED14 recruitment but not TF gene expression. Considering the essential role of MED14 in Mediator function, other coactivators may play major roles in HSP activation depending on the cellular conditions. Our findings highlight the importance of differential recruitment of Mediator for the precise control of HS responses in plants.

Comparative transcriptome analysis of Rosa chinensis ‘Slater's crimson China’ provides insights into the crucial factors and signaling pathways in heat stress response

Heat stress limits the growth of roses and adversely affects the yield and the quality of the rose cut-flowers. To investigate the heat stress response (HSR) mechanisms of rose, we compared the transcriptome profiling generated from Rosa chinensis ‘Slater's crimson China’ exposed to heat stress for five different time duration (0, 0.5, 2, 6, 12 h). Overall, 6175 differentially expressed genes (DGEs) were identified and exhibited different temporal expression patterns. Up-regulated genes related to chaperone-mediated protein folding, signal transduction and ROS scavenging were rapidly induced after 0.5–2 h of heat treatment, which provides evidence for the early adjustments of heat stress response in R. chinensis. While the down-regulated genes related to light reaction, sucrose biosynthesis, starch biosynthesis and cell wall biosynthesis were identified after as short as 6 h of heat stress, which indicated the ongoing negative effects on the physiology of R. chinensis. Using weighted gene co-expression network analysis, we found that different heat stress stages could be delineated by several modules. Based on integrating the transcription factors with upstream enriched DNA motifs of co-expressed genes in these modules, the gene regulation networks were predicted and several regulators of HSR were identified. Of particular importance was the discovery of the module associated with rapid sensing and signal transduction, in which numerous co-expressed genes related to chaperones, Ca2+ signaling pathways and transcription factors were identified. The results of this study provided an important resource for further dissecting the role of candidate genes governing the transcriptional regulatory network of HSR in Rose.

MicroRNA-guided regulation of heat stress response in wheat

BackgroundWith rising global temperature, understanding plants’ adaptation to heat stress has implications in plant breeding. MicroRNAs (miRNAs) are small, non-coding, regulatory RNAs guiding gene expression at the post-transcriptional level. In this study, small RNAs and the degradome (parallel analysis of RNA ends) of leaf tissues collected from control and heat-stressed wheat plants immediately at the end of the stress period and 1 and 4 days later were analysed.ResultsSequencing of 24 small RNA libraries produced 55.2 M reads while 404 M reads were obtained from the corresponding 24 PARE libraries. From these, 202 miRNAs were ascertained, of which mature miRNA evidence was obtained for 104 and 36 were found to be differentially expressed after heat stress. The PARE analysis identified 589 transcripts targeted by 84 of the ascertained miRNAs. PARE sequencing validated the targets of the conserved members of miRNA156, miR166 and miR393 families as squamosa promoter-binding-like, homeobox leucine-zipper and transport inhibitor responsive proteins, respectively. Heat stress responsive miRNA targeted superoxide dismutases and an array of homeobox leucine-zipper proteins, F-box proteins and protein kinases. Query of miRNA targets to interactome databases revealed a predominant association of stress responses such as signalling, antioxidant activity and ubiquitination to superoxide dismutases, F-box proteins, pentatricopeptide repeat-containing proteins and mitochondrial transcription termination factor-like proteins.ConclusionThe interlaced data set generated in this study identified and validated heat stress regulated miRNAs and their target genes associated with thermotolerance. Such accurate identification and validation of miRNAs and their target genes are essential to develop novel regulatory gene-based breeding strategies.

Pectin Methylesterases: Cell Wall Remodeling Proteins Are Required for Plant Response to Heat Stress.

Heat stress (HS) is expected to be of increasing worldwide concern in the near future, especially with regard to crop yield and quality as a consequence of rising or varying temperatures as a result of global climate change. HS response (HSR) is a highly conserved mechanism among different organisms but shows remarkable complexity and unique features in plants. The transcriptional regulation of HSR is controlled by HS transcription factors (HSFs) which allow the activation of HS-responsive genes, among which HS proteins (HSPs) are best characterized. Cell wall remodeling constitutes an important component of plant responses to HS to maintain overall function and growth; however, little is known about the connection between cell wall remodeling and HSR. Pectin controls cell wall porosity and has been shown to exhibit structural variation during plant growth and in response to HS. Pectin methylesterases (PMEs) are present in multigene families and encode isoforms with different action patterns by removal of methyl esters to influencing the properties of cell wall. We aimed to elucidate how plant cell walls respond to certain environmental cues through cell wall-modifying proteins in connection with modifications in cell wall machinery. An overview of recent findings shed light on PMEs contribute to a change in cell-wall composition/structure. The fine-scale modulation of apoplastic calcium ions (Ca2+) content could be mediated by PMEs in response to abiotic stress for both the assembly and disassembly of the pectic network. In particular, this modulation is prevalent in guard cell walls for regulating cell wall plasticity as well as stromal aperture size, which comprise critical determinants of plant adaptation to HS. These insights provide a foundation for further research to reveal details of the cell wall machinery and stress-responsive factors to provide targets and strategies to facilitate plant adaptation.

The Arabidopsis endoplasmic reticulum associated degradation pathways are involved in the regulation of heat stress response

The Cytosolic Protein Response (CPR) in the cytosol and the Unfolded Protein Response (UPR) and ER-associated degradation (ERAD) in the endoplasmic reticulum are major pathways of the cellular proteostasis network. However, despite years of effort, how these protein quality control systems coordinated invivo remains largely unknown, particularly in plants. In this study, the roles of two evolutionarily conserved ERAD pathways (DOA10 and HRD1) in heat stress response were investigated through reverse genetic approaches in Arabidopsis. Phenotypic analysis of the mutants showed that the two ERAD pathways additively play negative roles in heat tolerance, which was demonstrated by higher survival rate and lower electrolyte leakage in the loss of function mutants compared to the wild type plants. Importantly, gene expression analysis revealed that the mutant plants showed elevated transcriptional regulation of several downstream genes, including those encoding CPR and UPR marker genes, under both basal and heat stress conditions. Finally, multiple components of ERAD genes exhibited rapid response to increasing temperature. Taken together, our data not only unravels key insights into the crosstalk between different protein quality control processes, but also provides candidate genes to genetically improve plant heat tolerance in the future.

Arabidopsis heat stress transcription factors A2 (HSFA2) and A3 (HSFA3) function in the same heat regulation pathway

Heat stress transcription factors (HSFs) play an essential role in the adjustment of plants to high temperatures. These molecules have evolved complicated mechanisms that rely on interactions between different HSFs and other heat stress-related genes [such as bZIP28, multiprotein bridging factor 1c (MBF1c), calmodulin-binding protein kinase 3 (CBK3)] in response to different heat stresses (such as occasional or successive high temperatures). In the present study, phenotypic, gene expression and yeast two-hybrid assays revealed that HSFA2 and HSFA3 function in the same heat regulation pathway. The single mutants, hsfa2 and hsfa3 as well as double mutant hsfa2 and hsfa3, exhibited heat-sensitive phenotypes in acquired thermotolerance after a long recovery time (ATLR) but not in basic thermotolerance and acquired thermotolerance after a short recovery time (ATSR). The expression of HSP18.1-CI and HSP25.3-P was down-regulated in single and double mutants of hsfa2 and hsfa3 under successive heat stress in ATLR assays. In addition, HSFA2 interacted with HSFA3 at the protein level in yeast two-hybrid assays. These results demonstrated dynamic alterations in the expression of HSFA2, HSFA3 and other heat-related genes in ATLR assays, providing new insights into the relationship between HSFA2 and HSFA3; this information will refine the HSF network in the regulation of heat stress response.

Identification of SNP in HSP90AB1 and its Association with the Relative Thermotolerance and Milk Production Traits in Indian Dairy Cattle

Heat shock proteins (Hsp) play crucial role in cellular thermotolerance and heat stress response. In the present work, Allele specific PCR (AS-PCR) was standardized to detect the nucleotide polymorphism within the HSP90AB1 gene (SNP g.4338T>C) in Indian breeds of dairy cattle. The identified genotypes were associated with relative thermotolerance in terms of physiological parameters and milk production traits. The results of the experiments revealed that the genotype frequency of CC, CT, and TT for Sahiwal were 0.05, 0.78, and 0.17, respectively, and in Frieswal, the frequencies were 0.20, 0.70, and 0.10, respectively. The average rectal temperature (ART) and average respiration rates (ARR) were recorded during peak summer stress and heat tolerance coefficient (HTC) was calculated. The association studies indicated that TT genotypes had significantly (P < 0.01) higher HTC and lower ARR values than CT and CC in both the breeds. The TT genotype animals also had better production parameter in terms of total milk yield (TMY) (P < 0.01). These findings may partly suggest the role of HSP90AB1 polymorphisms in the regulation of heat stress response and consequent effect on production traits. Nevertheless, involvement of other regulatory mechanisms cannot be overruled.

Epigenetic regulation of heat stress response in plants

Owing to their sessile nature, plants have to passively respond to daily and seasonal temperature changes. Plants have evolved sophisticated genetic and epigenetic regulatory systems to perceive the ambient temperature and adjust the growth and development to survive. Global warming has posed a serious threat to the global food supply. It is of great urgency to investigate the molecular mechanism of heat stress response in plants. DNA methylation, histone modification, chromatin remodeling and small RNAs are major epigenetic regulation mechanisms. These epigenetic modifications have their own roles and work together in regulating the thermotolerance of plants. In this review, we summarized recent published researches on the epigenetic regulation of heat stress response in plants.