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Drought stress can seriously affect the yield and quality of wheat (Triticum aestivum). So far, although few wheat heat shock transcription factors (Hsfs) have been found to be involved in the stress response, the biological functions of them, especially the members of the HsfC (heat shock transcription factor C) subclass, remain largely unknown. Here, we identified a class C encoding gene, TaHsfC3-4, based on our previous omics data and analyzed its biological function in transgenic plants. TaHsfC3-4 encodes a protein containing 274 amino acids and shows the basic characteristics of the HsfC class. Gene expression profiles revealed that TaHsfC3-4 was constitutively expressed in many tissues of wheat and was induced during seed maturation. TaHsfC3-4 could be upregulated by PEG and abscisic acid (ABA), suggesting that this Hsf may be involved in the regulation pathway depending on ABA in drought resistance. Further results represented that TaHsfC3-4 was localized in the nucleus but had no transcriptional activation activity. Notably, overexpression of TaHsfC3-4 in Arabidopsis thaliana pyr1pyl1pyl2pyl4 (pyr1pyl124) quadruple mutant plants complemented the ABA-hyposensitive phenotypes of the quadruple mutant including cotyledon greening, root elongation, seedling growth, and increased tolerance to drought, indicating positive roles of TaHsfC3-4 in the ABA signaling pathway and drought tolerance. Furthermore, we identified TaHsfA2-11 as a TaHsfC3-4-interacting protein by yeast two-hybrid (Y2H) screening. The experimental data show that TaHsfC3-4 can indeed interact with TaHsfA2-11 in vitro and in vivo. Moreover, transgenic Arabidopsis TaHsfA2-11 overexpression lines exhibited enhanced drought tolerance, too. In summary, our study confirmed the role of TaHsfC3-4 in response to drought stress and provided a target locus for marker-assisted selection breeding to improve drought tolerance in wheat. [ABSTRACT FROM AUTHOR]

Heat shock transcription factor (Hsf) plays a pivotal role in promoting rapid heat‐induced transcriptional reprogramming in plants. The thermotolerance regulatory function of Hsfs is influenced by their own alternative splicing. In this study, we found that ZmHsf17‐II, an intron retention isoform of subclass A2 gene ZmHsf17 of maize (Zea mays), accumulated in large amounts as a result of severe or sustained heat stress. It was confirmed by expression and purification that ZmHsf17‐II encodes a small truncated peptide with 115 amino acids. ZmHsf17‐II was found to be located in the nucleus and have no transcriptional activity. Overexpressing ZmHsf17‐I in Arabidopsis could enhance plants' thermotolerance, while overexpressing ZmHsf17‐II does not. Based on the results of molecular docking, Y2H and split LUC experiments, we found that ZmHsf17‐II could bind to DBD region of ZmHsf17‐I through the hydrogen bond interaction between the truncated DBD of ZmHsf17‐II and three amino acid residues (Arg105, Thr109 and Lys142) of ZmHsf17‐I DBD region. Further experiments showed that ZmHsf17‐I could bind to its own promoter and exhibited transcriptional activation activity, while ZmHsf17‐I interaction with ZmHsf17‐II, transcriptional activation activity was interfered. Those findings indicate that ZmHsf17 can negatively regulate its own transcription by producing more intron retention isoforms via alternative splicing under heat stress. [ABSTRACT FROM AUTHOR]

High temperature has severely affected plant growth and development, resulting in reduced production of crops worldwide, especially wheat. Alternative splicing (AS), a crucial post-transcriptional regulatory mechanism, is involved in the growth and development of eukaryotes and the adaptation to environmental changes. Previous transcriptome data suggested that heat shock transcription factor (Hsf) TaHsfA2-7 may form different transcripts by AS. However, it remains unclear whether this post-transcriptional regulatory mechanism of TaHsfA2-7 is related to thermotolerance in wheat (Triticum aestivum). Here, we identified a novel splice variant, TaHsfA2-7-AS, which was induced by high temperature and played a positive role in thermotolerance regulation in wheat. Moreover, TaHsfA2-7-AS is predicted to encode a small truncated TaHsfA2-7 isoform, retaining only part of the DNA-binding domain (DBD). TaHsfA2-7-AS is constitutively expressed in various tissues of wheat. Notably, the expression level of TaHsfA2-7-AS is significantly up-regulated by heat shock (HS) during flowering and grain-filling stages in wheat. Further studies showed that TaHsfA2-7-AS was localized in the nucleus but lacked transcriptional activation activity. Ectopic expression of TaHsfA2-7-AS in yeast exhibited improved thermotolerance. Compared to non-transgenic plants, overexpression of TaHsfA2-7-AS in Arabidopsis results in enhanced tolerance to heat stress. Simultaneously, we also found that TaHsfA1 is directly involved in the transcriptional regulation of TaHsfA2-7 and TaHsfA2-7-AS. In summary, our findings demonstrate the function of TaHsfA2-7-AS splicing variant in response to heat stress and establish a link between regulatory mechanisms of AS and the improvement of thermotolerance in wheat. [ABSTRACT FROM AUTHOR]

Heat shock transcription factor (Hsf) exists widely in eukaryotes and responds to various abiotic stresses by regulating the expression of downstream transcription factors, functional enzymes, and molecular chaperones. In this study, TaHsfA2-13 , a heat shock transcription factor belonging to A2 subclass, was cloned from wheat (Triticum aestivum) and its function was analyzed. TaHsfA2-13 encodes a protein containing 368 amino acids and has the basic characteristics of Hsfs. Multiple sequence alignment analysis showed that TaHsfA2-13 protein had the highest similarity with TdHsfA2c-like protein from Triticum dicoccoides , which reached 100%. The analysis of tissue expression characteristics revealed that TaHsfA2-13 was highly expressed in root, shoot, and leaf during the seedling stage of wheat. The expression of TaHsfA2-13 could be upregulated by heat stress, low temperature, H2O2, mannitol, salinity and multiple phytohormones. The TaHsfA2-13 protein was located in the nucleus under the normal growth conditions and showed a transcriptional activation activity in yeast. Further studies found that overexpression of TaHsfA2-13 in Arabidopsis thaliana Col-0 or athsfa2 mutant results in improved tolerance to heat stress, H2O2, SA and mannitol by regulating the expression of multiple heat shock protein (Hsp) genes. In summary, our study identified TaHsfA2-13 from wheat, revealed its regulatory function in varieties of abiotic stresses, and will provide a new target gene to improve stress tolerance for wheat breeding. [ABSTRACT FROM AUTHOR]

High temperature is a major constrait limiting the yield and quality of wheat. Wheat heat shock transcription factor (Hsf) plays important roles in regulating plant response to heat shock. In previous study, we found there are 82 Hsfs in wheat and TaHsfA2–11 expression was obviously upregulated by heat stress. In this study, we aim to investigate TaHsfA2–11 function and regulating mechanism in response to heat shock through genetic transformation into wheat Fielder. Gene expression analyses results showed that TaHsfA2–11 was expressed in all detected tissues and the most highly expressed level was in wheat mature roots. The expression level of TaHsfA2–11 was strongly upregulated by heat shock in wheat leaves. Subcellular localization results represented that TaHsfA2–11 is located to the nucleus. Transgenic wheat seedlings overexpressing TaHsfA2–11 showed stronger thermotolerance than the control Fielder after treated under 50 °C for 12 h. After heat shock, the TaHsfA2–11 overexpressing lines showed increased survival rate, chlorophyll content, POD (Peroxidase) activity, SOD (Superoxide dismutase) activity and photosynthetic rate, and decreased relative electrolyte leakage and MDA (malondialdehyde) content compared with Fielder. The results of RNA-sequence data demonstrated that TaHsfA2–11 can regulate many heat response genes, such as Hsp genes TaHsp16.6 and TaHsp21 , ER (Endoplasmic reticulum) stress genes TaBiP2 and TaERDJ3A, photoprotectant for PSII gene TaELIPHV58 , flavone biosynthesis related genes TaUGT-2A and TaPMaT , etc after heat stress. These results showed that TaHsfA2–11 can impove thermotolerance perhaps through effecting signal pathway of heat, ER stress, photosynthesis and ROS (Reactive Oxygen Species). The findings will provide useful gene resource for performing genetic modification of wheat thermotolerance in heat-resistant breeding. • TaHsfA2-11 up-regulated in wheat response to HS, H 2 O 2 and SA treatment. • TaHsfA2-11 has transactivation transcriptional activity. • Overexpression of TaHsfA2-11 in wheat improves thermotolerance. [ABSTRACT FROM AUTHOR]