Functions of Phytochrome-Interacting Factors (PIFs) in the regulation of plant growth and development: A comprehensive review.

Transcription factors play important roles in governing plant responses upon changes in their ambient conditions. Any fluctuation in the supply of critical requirements for plants, such as optimum light, temperature, and water leads to the reprogramming of gene-signaling pathways. At the same time, plants also evaluate and shift their metabolism according to the various stages of development. Phytochrome-Interacting Factors are one of the most important classes of transcription factors that regulate both developmental and external stimuli-based growth of plants. This review focuses on the identification of PIFs in various organisms, regulation of PIFs by various proteins, functions of PIFs of Arabidopsis in diverse developmental pathways such as seed germination, photomorphogenesis, flowering, senescence, seed and fruit development, and external stimuli-induced plant responses such as shade avoidance response, thermomorphogenesis, and various abiotic stress responses. Recent advances related to the functional characterization of PIFs of crops such as rice, maize, and tomato have also been incorporated in this review, to ascertain the potential of PIFs as key regulators to enhance the agronomic traits of these crops. Thus, an attempt has been made to provide a holistic view of the function of PIFs in various processes in plants.

Ectopic overexpression of TaHsfA5 promotes thermomorphogenesis in Arabidopsis thaliana and thermotolerance in Oryza sativa.

Heat stress transcription factors (Hsfs) play an important role in regulating the heat stress response in plants. Among the Hsf family members, the group A members act upstream in initiating the response upon sensing heat stress and thus, impart thermotolerance to the plants. In the present study, wheat HsfA5 (TaHsfA5) was found to be one of the Hsfs, which was upregulated both in heat stress and during the recovery period after the stress. TaHsfA5 was found to interact with TaHsfA3 and TaHsfA4, both of which are known to positively regulate the heat stress-responsive genes. Apart from these, TaHsfA5 also interacted with TaHSBP2 protein, whose role has been implicated in attenuating the heat stress response. Further, its heterologous overexpression in Arabidopsis and Oryza sativa promoted thermotolerance in these plants. This indicated that TaHsfA5 positively regulated the heat stress response. Interestingly, the TaHsfA5 overexpression Arabidopsis plants when grown at warm temperatures showed  a hyper-thermomorphogenic response in comparison to the wild-type plants. This was found to be consistent with the higher expression of PIF4 and its target auxin-responsive genes in these transgenics in contrast to the wild-type plants. Thus, these results suggest the involvement of TaHsfA5 both in the heat stress response as well as in the thermomorphogenic response in plants.

The heat stress transcription factor family in Aegilops tauschii: genome-wide identification and expression analysis under various abiotic stresses and light conditions.

Heat stress transcription factors (Hsfs) are known to play a vital role in protecting plants against various abiotic stresses. Among the wild wheat relatives, Aegilops tauschii offers an excellent source of abiotic stress tolerance genes for improvement of bread wheat. However, little is known about its stress tolerance mechanisms. In this study, 22 AetHsf genes were identified in the genome of Aegilops tauschii and their chromosomal location, exon-intron structures, sub-cellular localization, phylogenetic and syntenic relationship were analyzed. Based on the conserved motif analysis, these Hsfs were further divided into group A, B and C. The interaction network analysis and expression profile of AetHsfs in different tissues predicted their interaction with diverse types of proteins and suggested their involvement in different developmental processes of the plant. The promoter analysis of AetHsfs showed the presence of abiotic stress-responsive, phytohormone-responsive, plant development-related and light-related cis-elements. Thus, we investigated the expression of Hsfs in Aegilops tauchii seedlings under various abiotic stress conditions and irradiated with different monochromatic lights. Most of the AetHsfs were found to be upregulated by heat stress, while some showed expression in drought, salinity and high light stress as well. Notably, the expression pattern of various AetHsfs showed their responsiveness toward dark and various light conditions (blue red and far-red) as well. Thus, this study provides novel insights into the potential role of AetHsfs in stress and light signaling pathways, which can further facilitate understanding of the stress tolerance mechanisms in Aegilops tauschii.

Identification and Characterization of DNA Demethylase Genes and Their Association With Thermal Stress in Wheat (Triticum aestivum L.).

DNA demethylases (dMTases) are essential proteins in plants that regulate DNA methylation levels. The dMTase genes have been explored in a number of plant species, however, members of this family have not been reported in wheat. We identified 12 wheat dMTase genes divided into two subfamilies: repressor of silencing 1 (ROS1) and DEMETER-Like (DML). The TadMTases in the same subfamily or clade in the phylogenetic tree have similar gene structures, protein motifs, and domains. The promoter sequence contains multiple cis-regulatory elements (CREs) that respond to abiotic stress, hormones, and light, suggesting that the majority of TadMTase genes play a role in wheat growth, development, and stress response. The nuclear localization signals (NLSs), subcellular localization, and SRR motifs were also analyzed. The expression profile analyses revealed that TadMTase genes showed differential gene expression patterns in distinct developmental stages and tissues as well as under heat stress (HS). Furthermore, the qRT-PCR analysis revealed that TadMTase gene expression differed amongst wheat cultivars with varying degrees of HS tolerance. Overall, this work contributes to the understanding of the biological function of wheat dMTases and lays the foundation for future investigations.

Wheat ocs-Element Binding Factor 1 Enhances Thermotolerance by Modulating the Heat Stress Response Pathway.

The basic leucine zipper family (bZIP) represents one of the largest families of transcription factors that play an important role in plant responses to abiotic stresses. However, their role in contributing to thermotolerance in plants is not well explored. In this article, two homoeologs of wheat ocs-element binding factor 1 (TaOBF1-5B and TaOBF1-5D) were found to be heat-responsive TabZIP members. Their expression analysis in Indian wheat cultivars revealed their differential expression pattern and TaOBF1-5B was found to be more receptive to heat stress. Consistent with this, the heterologous overexpression of TaOBF1-5B in Arabidopsis thaliana and Oryza sativa promoted the expression of stress-responsive genes, which contributed to thermotolerance in transgenic plants. TaOBF1-5B was seen to interact with TaHSP90 in the nucleus and TaSTI in the nucleolus and the ER. Thus, the results suggest that TaOBF1-5B might play an important regulatory role in the heat stress response and is a major factor governing thermotolerance in plants.

Microbial Mechanisms of Heat Sensing.

Temperature is one of the ubiquitous signals that control both the development as well as virulence of various microbial species. Therefore their survival is dependent upon initiating appropriate response upon temperature fluctuations. In particular, pathogenic microbes exploit host-temperature sensing mechanisms for triggering the expression of virulence genes. Many studies have revealed that the biomolecules within a cell such as DNA, RNA, lipids and proteins help in sensing change in temperature, thereby acting as thermosensors. This review shall provide an insight into the different mechanisms of thermosensing and how they aid pathogenic microbes in host invasion.

Overexpression of HVA1 Enhances Drought and Heat Stress Tolerance in Triticum aestivum Doubled Haploid Plants.

Plant responses to multiple environmental stresses include various signaling pathways that allow plant acclimation and survival. Amongst different stresses, drought and heat stress severely affect growth and productivity of wheat. HVA1, a member of the group 3 LEA protein, has been well known to provide protection against drought stress. However, its mechanism of action and its role in other stresses such as heat remain unexplored. In this study, doubled haploid (DH) wheat plants overexpressing the HVA1 gene were analyzed and found to be both drought-and heat stress-tolerant. The transcriptome analysis revealed the upregulation of transcription factors such as DREB and HsfA6 under drought and heat stress, respectively, which contribute toward the tolerance mechanism. Particularly under heat stress conditions, the transgenic plants had a lower oxidative load and showed enhanced yield. The overexpression lines were found to be ABA-sensitive, therefore suggesting the role of HsfA6 in providing heat tolerance via the ABA-mediated pathway. Thus, apart from its known involvement in drought stress, this study highlights the potential role of HVA1 in the heat stress signaling pathway. This can further facilitate the engineering of multiple stress tolerance in crop plants, such as wheat.

Elucidating the functional role of heat stress transcription factor A6b (TaHsfA6b) in linking heat stress response and the unfolded protein response in wheat.

KEY MESSAGE: TaHsfA6b-4D relocalizes intracellularly upon heat stress and play a significant role in linking the heat stress response to unfolded-protein response so as to maintain cellular homeostasis. Heat stress transcription factors (Hsfs) play a crucial role in protecting the plants against heat stress (HS). In case of wheat, TaHsfA6b-4D (earlier known as TaHsfA2d) has been identified as a seed preferential transcription factor and its role has been shown in various abiotic stresses such as heat, salt and drought stress. In the present study, a homeologue of TaHsfA6b gene (TaHsfA6b-4A) was identified and was found to be transcriptionally inactive but it localized to the nucleus. Interestingly, TaHsfA6b-4D localized to the endoplasmic reticulum-Golgi complex and peroxisomes under non-stress conditions, but was observed to accumulate in the nucleus upon HS. The expression of TaHsfA6b-4D was upregulated by dithiothreitol (DTT), which is a known ER stress inducer. Consistent with this, Arabidopsis transgenic plants overexpressing TaHsfA6b-4D performed better on DTT containing media, which further corroborated with the increased expression of ER stress marker genes in these transgenic plants in comparison to the wild type plants. Thus, these studies together suggest that TaHsfA6b-4D may relocalize intracellularly upon heat stress and may play a significant role in linking the unfolded-protein response with heat stress response so as to maintain protein homeostasis inside the cell under heat stress.

Genome-wide identification and expression profiling of cytosine-5 DNA methyltransferases during drought and heat stress in wheat (Triticum aestivum).

DNA methylation is a potential epigenetic mechanism that regulates genome stability, development, and stress mitigation in plants. It is mediated by cytosine-5 DNA methyltransferases (C5-MTases). We identified 52 wheat C5-MTases; and based on domain structure and phylogenetics, these 52 C5-MTases were classified into four sub-families including MET, CMT, DRM and DNMT2; and were distributed on 18 chromosomes. Cis-acting regulatory elements analysis identified abiotic stress-responsive, phytohormone-responsive, development-related and light-related elements in the promoters of TaC5-MTases. We also examined the transcript abundance of TaC5-MTases in different tissues, developmental stages and under abiotic stresses. Notably, most of the TaC5-MTases (TaCMT2, TaCMT3b, TaCMT3c, TaMET1, TaDRM10, TaDNMT2) showed differential regulation of their transcript abundance during drought and heat stress. Overall, the above results provide significant insights into the expression and the probable functions of TaC5-MTases and will also expedite future research programs to explore the mechanisms of epigenetic regulation in wheat.

Dissecting the Molecular Function of Triticum aestivum STI Family Members Under Heat Stress.

STI/HOP functions as a co-chaperone of HSP90 and HSP70 whose molecular function has largely been being restricted as an adaptor protein. However, its role in thermotolerance is not well explored. In this article, we have identified six members of the TaSTI family, which were named according to their distribution on group 2 and group 6 chromosomes. Interestingly, TaSTI-2 members were found to express higher as compared to TaSTI-6 members under heat stress conditions, with TaSTI-2A being one of the most heat-responsive member. Consistent with this, the heterologous expression of TaSTI-2A in Arabidopsis resulted in enhanced basal as well as acquired thermotolerance as revealed by the higher yield of the plants under stress conditions. Similarly in the case of rice, TaSTI-2A transgenics exhibited enhanced thermal tolerance. Moreover, we demonstrate that TaSTI-2A interacts with TaHSP90 not only in the nucleus but also in the ER and Golgi bodies, which has not been shown till now. Additionally, TaHSP70 was also found to interact with TaSTI-6D specifically in the cytosol. Thus, these data together suggested that the TaSTI family members might play different roles under heat stress conditions in order to fine-tune the heat stress response in plants.

VDAC and its interacting partners in plant and animal systems: an overview.

Molecular trafficking between different subcellular compartments is the key for normal cellular functioning. Voltage-dependent anion channels (VDACs) are small-sized proteins present in the outer mitochondrial membrane, which mediate molecular trafficking between mitochondria and cytoplasm. The conductivity of VDAC is dependent on the transmembrane voltage, its oligomeric state and membrane lipids. VDAC acts as a convergence point to a diverse variety of mitochondrial functions as well as cell survival. This functional diversity is attained due to their interaction with a plethora of proteins inside the cell. Although, there are hints toward functional conservation/divergence between animals and plants; knowledge about the functional role of the VDACs in plants is still limited. We present here a comparative overview to provide an integrative picture of the interactions of VDAC with different proteins in both animals and plants. Also discussed are their physiological functions from the perspective of cellular movements, signal transduction, cellular fate, disease and development. This in-depth knowledge of the biological importance of VDAC and its interacting partner(s) will assist us to explore their function in the applied context in both plant and animal.

Arabidopsis Mitochondrial Voltage-Dependent Anion Channels Are Involved in Maintaining Reactive Oxygen Species Homeostasis, Oxidative and Salt Stress Tolerance in Yeast.

Voltage-dependent anion channels (VDACs) are conserved proteins of the mitochondria. We have functionally compared Arabidopsis VDACs using Saccharomyces cerevisiae Δpor1 and M3 yeast system. VDAC (1, 2, and 4) were able to restore Δpor1 growth in elevated temperature, in oxidative and salt stresses, whereas VDAC3 only partially rescued Δpor1 in these conditions. The ectopic expression of VDAC (1, 2, 3, and 4) in mutant yeast recapitulated the mitochondrial membrane potential thus, enabled it to maintain reactive oxygen species homeostasis. Overexpression of these VDACs (AtVDACs) in M3 strain did not display any synergistic or antagonistic activity with the native yeast VDAC1 (ScVDAC1). Collectively, our data suggest that Arabidopsis VDACs are involved in regulating respiration, reactive oxygen species homeostasis, and stress tolerance in yeast.

Alternative Splicing of CIPK3 Results in Distinct Target Selection to Propagate ABA Signaling in Arabidopsis.

Calcium (Ca2+) signaling is pivotal in transmission of information in the cell. Various Ca2+ sensing molecules work to sense and relay the encrypted messages to the intended targets in the cell to maintain this signal transduction. CBL-interacting protein kinases (CIPKs) are crucial components of Ca2+ signal transduction during various abiotic stresses. Although there are intron rich CIPKs in the plant genome but very little has been reported about their alternative splicing. Moreover the physiological significance of this event in the Ca2+ signaling is still elusive. Therefore in this study, we have selected CIPK3, which has highest number of splice variants amongst Arabidopsis CIPKs. Expression profiling of five splice variants of CIPK3 by qRT-PCR in four Arabidopsis thaliana ecotypes revealed preferential transcript accumulation but similar subcellular localization of the variants and interaction with similar CBLs. ABA and drought treatment resulted in the higher accumulation of the alternately spliced transcripts of CIPK3 in Arabidopsis ecotype Wassilewkija. The transcripts of CIPK3.1 and CIPK3.4 are relatively more induced compared to other alternative splice variants. Out of four splice variants studied, we found CIPK3.1 and CIPK3.2 showing preference for ABR1, a previously reported interactor of CIPK3. We conclude that the differential expression and choice of downstream partner by CIPK3-splice variants might be one of the mechanisms of Ca2+ mediated preferential regulation of ABA and other stress signals.