Regulator Of Drought Stress Response Research Articles

MdTPR16, an apple tetratricopeptide repeat (TPR)-like superfamily gene, positively regulates drought stress in apple

The Tetratricopeptide repeat (TPR)-like superfamily with TPR conserved domains is widely involved in the growth and abiotic stress in many plants. In this report, the gene MdTPR16 belongs to the TPR family in apple (Malus domestica). Promoter analysis reveal that MdTPR16 incorporated various stress response elements, including the drought stress response elements. And different abiotic stress treatments, drought especially, significantly induce the response of MdTPR16. Overexpression of MdTPR16 result in better drought tolerance in apple and Arabidopsis by up-regulating the expression levels of drought stress-related genes, achieving a higher chlorophyll content level, more material accumulation, and overall better growth compared to WT in the drought. Under drought stress, the overexpressed MdTPR16 also mitigate the oxidative damage in cells by reducing the electrolyte leakage, malondialdehyde content, and the H2O2 and O2− accumulation in apples and Arabidopsis. In conclusion, MdTPR16 act as a beneficial regulator of drought stress response by regulating the expression of related genes and the cumulation of reactive oxygen species (ROS).

The SPL transcription factor TaSPL6 negatively regulates drought stress response in wheat

Environmental stresses, such as heat and drought, severely affect plant growth and development, and reduce wheat yield and quality globally. Squamosa promoter binding protein-like (SPL) proteins are plant-specific transcription factors that play a critical role in regulating plant responses to diverse stresses. In this study, we cloned and characterized TaSPL6, a wheat orthologous gene of rice OsSPL6. Three TaSPL6 homoeologs are located on the long arms of chromosomes 4A, 5B, and 5D, respectively, and share more than 98% sequence identity with each other. The TaSPL6 genes were preferentially expressed in roots, and their expression levels were downregulated in wheat seedlings subjected to heat, dehydration, and abscisic acid treatments. Subcellular localization experiments showed that TaSPL6 was localized in the nucleus. Overexpression of TaSPL6-A in wheat resulted in enhanced sensitivity to drought stress. The transgenic lines exhibited higher leaf water loss, malondialdehyde and reactive oxygen species (ROS) content, and lower antioxidant enzyme activities after drought treatment than wild-type plants. Gene silencing of TaSPL6 enhanced the drought tolerance of wheat, as reflected by better growth status. Additionally, RNA-seq and qRT-PCR analyses revealed that TaSPL6-A functions by decreasing the expression of a number of genes involved in stress responses. These findings suggest that TaSPL6 acts as a negative regulator of drought stress responses in plants, which may have major implications for understanding and enhancing crop tolerance to environmental stresses.

DEAD-box RNA helicase 6 regulates drought and abscisic acid stress responses in rapeseed (Brassica napus)

DEAD-box RNA helicase is a major subfamily of RNA helicases with vital roles played in plant growth, development, and plant-environment interactions. RNA helicase 6 in rapeseed (Brassica napus) (BnRH6) is a member of DEAD-box RNA helicase. While previous research has demonstrated the role of BnRH6 in salt stress regulation, the involvement of BnRH6 in drought stress adaptation remains unknown. This report described a function of BnRH6 in drought stress response. BnRH6 was sufficiently induced by osmotic stress. Transgenic Brassica napus and Arabidopsis thaliana (Arabidopsis) overexpressing BnRH6 (OE) showed a drought tolerance phenotype, characterized by improved plant growth, increased survival rates, reduced water loss, leaf chlorosis and oxidative stress. Furthermore, BnRH6 was also induced by exogenous abscisic acid (ABA). BnRH6 overexpression plants exhibited ABA hypersensitivity with lagging seed germination, growth stunt and diminished stomatal opening in the presence of ABA, suggesting the involvement of ABA signal. Assessment of several well-identified drought stress responsive genes such as Calcium-dependent Protein Kinase 14 (BnCDPK14), Enhanced Response to ABA1 (BnERA1) and ABA Insensitive 1 (BnABI1) revealed that their expressions were accordingly changed in BnOE plants, possibly with interplays between BnRH6 and those genes. Our data highlighted the functional role of BnRH6, which plays a positive role in active regulation of drought stress response likely through an ABA-dependent manner in rapeseed plants.

Pepper homeobox abscisic acid signalling-related transcription factor 1, CaHAT1, plays a positive role in drought response.

Abscisic acid (ABA) signalling triggers drought resistance mediated by SNF1-related kinase 2s (SnRK2s), which transmits stress signals through the phosphorylation of several downstream factors. However, these kinases and their downstream targets remain elusive in pepper plants. This study aimed to isolate interacting partners of CaSnRK2.6, a homologue of Arabidopsis SnRK2.6/OST1. Among the candidate proteins, we identified a homeodomain-leucine zipper (HD-Zip) class II protein and named it CaHAT1 (Capsicum annuum homeobox ABA signalling related- transcription factor 1). CaHAT1-silenced pepper and -overexpression (OE) transgenic Arabidopsis plants were generated to investigate the in vivo function of CaHAT1 in drought response. Following the application of drought stress, CaHAT1-silenced pepper plants exhibited drought-sensitive phenotypes with reduced ABA-mediated stomatal closure and lower expression of stress-responsive genes compared with control plants. In contrast, CaHAT1-OE transgenic Arabidopsis plants showed the opposite phenotypes, including increased drought resistance and ABA sensitivity. CaHAT1, particularly its N-terminal consensus sequences, was directly phosphorylated by CaSnRK2.6. Furthermore, CaSnRK2.6 kinase activity and CaSnRK2.6-mediated CaHAT1 phosphorylation levels were enhanced by treatment with ABA and drought stress. Taken together, our results indicated that CaHAT1, which is the target protein of CaSnRK2.6, is a positive regulator of drought stress response. This study advances our understanding of CaHAT1-CaSnRK2.6 mediated defence mechanisms in pepper plants against drought stress.

The Arrangement of MicroRNAs in the Regulation of Drought Stress Response in Plants: A Systematic Review

The Arrangement of MicroRNAs in the Regulation of Drought Stress Response in Plants: A Systematic Review

OsWR2 recruits HDA704 to regulate the deacetylation of H4K8ac in the promoter of OsABI5 in response to drought stress.

Drought stress is a major environmental factor that limits the growth, development, and yield of rice (Oryza sativa L.). Histone deacetylases (HDACs) are involved in the regulation of drought stress responses. HDA704 is an RPD3/HDA1 class HDAC that mediates the deacetylation of H4K8 (lysine 8 of histone H4) for drought tolerance in rice. In this study, we show that plants overexpressing HDA704 (HDA704-OE) are resistant to drought stress and sensitive to abscisic acid (ABA), whereas HDA704 knockout mutant (hda704) plants displayed decreased drought tolerance and ABA sensitivity. Transcriptome analysis revealed that HDA704 regulates the expression of ABA-related genes in response to drought stress. Moreover, HDA704 was recruited by a drought-resistant transcription factor, WAX SYNTHESIS REGULATORY 2 (OsWR2), and co-regulated the expression of the ABA biosynthesis genes NINE-CIS-EPOXYCAROTENOID DIOXYGENASE 3 (NCED3), NCED4, and NCED5 under drought stress. HDA704 also repressed the expression of ABA-INSENSITIVE 5 (OsABI5) and DWARF AND SMALL SEED 1 (OsDSS1) by regulating H4K8ac levels in the promoter regions in response to polyethylene glycol 6000 treatment. In agreement, the loss of OsABI5 function increased resistance to dehydration stress in rice. Our results demonstrate that HDA704 is a positive regulator of the drought stress response and offers avenues for improving drought resistance in rice.

The clade F PP2C phosphatase ZmPP84 negatively regulates drought tolerance by repressing stomatal closure in maize.

Drought is a major environmental stress that threatens crop production. Therefore, identification of genes involved in drought stress response is of vital importance to decipher the molecular mechanism of stress signal transduction and breed drought tolerance crops, especially for maize. Clade A PP2C phosphatases are core abscisic acid (ABA) signaling components, regulating ABA signal transduction and drought response. However, the roles of other clade PP2Cs in drought resistance remain largely unknown. Here, we discovered a clade F PP2C, ZmPP84, that negatively regulates drought tolerance by screening a transgenic overexpression maize library. Quantitative RT-PCR indicates that the transcription of ZmPP84 is suppressed by drought stress. We identified that ZmMEK1, a member of the MAPKK family, interacts with ZmPP84 by immunoprecipitation and mass spectrometry analysis. Additionally, we found that ZmPP84 can dephosphorylate ZmMEK1 and repress its kinase activity on the downstream substrate kinase ZmSIMK1, while ZmSIMK1 is able to phosphorylate S-type anion channel ZmSLAC1 at S146 and T520 invitro. Mutations of S146 and T520 to phosphomimetic aspartate could activate ZmSLAC1 currents in Xenopus oocytes. Taken together, our study suggests that ZmPP84 is a negative regulator of drought stress response that inhibits stomatal closure through dephosphorylating ZmMEK1, thereby repressing ZmMEK1-ZmSIMK1 signaling pathway.

Basic helix-loop-helix transcription factor PxbHLH02 enhances drought tolerance in Populus (Populus simonii × P. nigra).

The basic helix-loop-helix (bHLH) transcription factors (TFs) are involved in plant morphogenesis and various abiotic and biotic stress responses. However, further exploration is required of drought-responsive bHLH family members and their detailed regulatory mechanisms in Populus. Two bHLH TF genes, PxbHLH01/02, were identified in Populus simonii × P. nigra and cloned. The aim of this study was to examine the role of bHLH TFs in drought tolerance in P. simonii × P. nigra. The results showed that the amino acid sequences of the two genes were homologous to Arabidopsis thaliana UPBEAT1 (AtUPB1) and overexpression of PxbHLH01/02 restored normal root length in the AtUPB1 insertional mutant (upb1-1). The PxbHLH01/02 gene promoter activity analysis suggested that they were involved in stress responses and hormonesignaling. Furthermore, Arabidopsis transgenic lines overexpressing PxbHLH01/02 exhibited higher stress tolerance compared with the wild-type. Populus simonii × P. nigra overexpressing PxbHLH02 increased drought tolerance and exhibited higher superoxide dismutase and peroxidase activities, lower H2O2and malondialdehyde content, and lower relative conductivity. The results of transcriptome sequencing (RNA-seq) and quantitative real-time PCR suggested that the response of PxbHLH02 to drought stress was related to abscisicacid (ABA) signal transduction. Overall, the findings of this study suggest that PxbHLH02 from P. simonii × P. nigra functions as a positive regulator of drought stress responses by regulating stomatal aperture and promoting ABA signal transduction.

Molecular Characterization of U-box E3 Ubiquitin Ligases (TaPUB2 and TaPUB3) Involved in the Positive Regulation of Drought Stress Response in Arabidopsis.

Plant U-box E3 ubiquitin ligase (PUB) is involved in various environmental stress conditions. However, the molecular mechanism of U-box proteins in response to abiotic stress in wheat remains unknown. In this study, two U-box E3 ligase genes (TaPUB2 and TaPUB3), which are highly expressed in response to adverse abiotic stresses, were isolated from common wheat, and their cellular functions were characterized under drought stress. Transient expression assay revealed that TaPUB2 was localized in the cytoplasm and Golgi apparatus, whereas TaPUB3 was expressed only in the Golgi apparatus in wheat protoplasts. Additionally, TaPUB2 and TaPUB3 underwent self-ubiquitination. Moreover, TaPUB2/TaPUB3 heterodimer was identified in yeast and the cytoplasm of wheat protoplasts using a pull-down assay and bimolecular fluorescence complementation analysis. Heterogeneous overexpression of TaPUB2 and TaPUB3 conferred tolerance to drought stress. Taken together, these results implied that the heterodimeric form of U-box E3 ubiquitin ligases (TaPUB2/TaPUB3) responded to abiotic stress and roles as a positive regulator of drought stress tolerance.

OsPUB41, a U-box E3 ubiquitin ligase, acts as a negative regulator of drought stress response in rice (Oryza Sativa L.).

OsPUB41 plays a negative role in drought stress response through the mediation of OsUBC25 and interacts with OsCLC6, suggesting a putative substrate. The notable expansion of Plant U-Box E3 ligases (PUB), compared with those in mammals, implies that PUB proteins have evolved to perform plant-specific functions. OsPUB41, a potential ortholog of CMPG1, was recently reported to regulate the cell wall degrading enzyme (CWDE)-induced innate immune response in rice. Here, we characterized the OsPUB41 gene, which encodes a dual-localized cytosolic and nuclear U-box E3 ligase in rice. OsPUB41 expression was specifically induced by dehydration among various abiotic stresses and abscisic acid (ABA) treatments. Furthermore, we revealed that the core U-box motif of OsPUB41 possesses the E3 ligase activity that can be activated by OsUBC25 in rice. The Ubi:RNAi-OsPUB41 knock-down and ospub41 suppression mutant plants exhibited enhanced tolerance to drought stress compared with the wild-type rice plants in terms of transpirational water loss, long-term dehydration response, and chlorophyll content. Moreover, the knock-down or suppression of the OsPUB41 gene did not cause adverse effect onrice yield-related traits. Yeast two-hybrid and an in vitro pull-down analyses revealed that OsCLC6, a chloride channel, is a putative substrate of OsPUB41. Overall, these results suggest that OsPUB41 acts as a negative regulator of dehydration conditions and interacts with OsCLC6, implying that it is a substrate of OsPUB41.

Current status of microRNA-mediated regulation of drought stress responses in cereals.

Drought is one of the most important abiotic stress factors impeding crop productivity. With the uncovering of their role as potential regulators of gene expression, microRNAs (miRNAs) have been recognized as new targets for developing stress resistance. MicroRNAs are small noncoding RNAs whose abundance is significantly altered under stress conditions. Interestingly, plant miRNAs predominantly targets transcription factors (TFs), and some of which are also the most critical drought-responsive genes that in turn could regulate the expression of numerous loci with drought-adaptive potential. The phytohormone ABA plays important roles in regulating stomatal conductance and in initiating an adaptive response to drought stress. miRNAs are implicated in regulating ABA-(abscisic acid) and non-ABA-mediated drought resistance pathways. For instance, miR159-MYB module and miR169-NFYA module participates in an ABA-dependent pathway, whereas several other ABA-independent miRNA-target modules (miR156-SPL; miR393-TIR1; miR160-ARF10, ARF16, ARF17; miR167-ARF6 and ARF8; miR390/TAS3siRNA-ARF2, ARF3, ARF4) collectively regulate drought responses in plants. Overall, miRNA-mediated drought response manifests diverse molecular, biochemical and physiological processes. Because of their immense role in controlling gene expression, miRNA manipulation has significant potential to augment plant tolerance to drought stress. This review compiles the current understanding of drought-responsive miRNAs in major cereals. Also, potential miRNA manipulation strategies currently in use along with the challenges and future perspectives are discussed.

Mitogen-Activated Protein Kinase CaDIMK1 Functions as a Positive Regulator of Drought Stress Response and Abscisic Acid Signaling in Capsicum annuum.

Protein phosphorylation by kinase is an important mechanism for adapting to drought stress conditions. Here, we isolated the CaDIMK1 (Capsicum annuum drought-induced MAP kinase 1) from dehydrated pepper leaf tissue and functionally characterized it. Subcellular localization analysis revealed that the CaDIMK1 protein was localized in the cytoplasm and nucleus. CaDIMK1-silenced pepper plants exhibited drought-susceptible phenotypes that were characterized by increased transpiration rates, low leaf temperatures, and decreased stomatal closure. In contrast, CaDIMK1-overexpressing (OX) transgenic Arabidopsis plants were hypersensitive to abscisic acid (ABA) from germination to adult growth stages. Furthermore, the CaDIMK1-OX plants were tolerant to drought stress. The transcript levels of several stress-related genes were high in CaDIMK1-OX plants than in wild-type plants. Taken together, our data demonstrate that CaDIMK1 acts as a positive modulator of drought tolerance and ABA signal transduction in pepper plants.

Hydrogen sulfide: An emerging signaling molecule regulating drought stress response in plants.

Hydrogen sulfide (H2 S) is a small, reactive signaling molecule that is produced within chloroplasts of plant cells as an intermediate in the assimilatory sulfate reduction pathway by the enzyme sulfite reductase. In addition, H2 S is also produced in cytosol and mitochondria by desulfhydration of l-cysteine catalyzed by l-cysteine desulfhydrase (DES1) in the cytosol and from β-cyanoalanine in mitochondria, in a reaction catalyzed by β-cyano-Ala synthase C1 (CAS-C1). H2 S exerts its numerous biological functions by post-translational modification involving oxidation of cysteine residues (RSH) to persulfides (RSSH). At lower concentrations (10-1000 μmol L-1 ), H2 S shows huge agricultural potential as it increases the germination rate, the size, fresh weight, and ultimately the crop yield. It is also involved in abiotic stress response against drought, salinity, high temperature, and heavy metals. H2 S donor, for example, sodium hydrosulfide (NaHS), has been exogenously applied on plants by various researchers to provide drought stress tolerance. Exogenous application results in the accumulation of polyamines, sugars, glycine betaine, and enhancement of the antioxidant enzyme activities in response to drought-induced osmotic and oxidative stress, thus, providing stress adaptation to plants. At the biochemical level, administration of H2 S donors reduces malondialdehyde content and lipoxygenase activity to maintain the cell integrity, causes abscisic acid-mediated stomatal closure to prevent water loss through transpiration, and accelerates the photosystem II repair cycle. Here, we review the crosstalk of H2 S with secondary messengers and phytohormones towards the regulation of drought stress response and emphasize various approaches that can be addressed to strengthen research in this area.

Interference of CsGPA1, the α-submit of G protein, reduces drought tolerance in cucumber seedlings

The G protein α-subunit, GPA1, is an integral component of several signaling pathways in plants, including response to abiotic stress. However, the molecular mechanism behind these processes remains largely unknown in the cucumber plant (Cucumis sativus L.). In order to further understand the role of CsGPA1 in cucumber under drought stress, changes in plant growth, physiological parameters, and gene expression of CsAQPs were all measured under water stress induced by polyethylene glycol (PEG) using wild type (WT) and CsGPA1-interference (RNAi) cucumber seedlings. Our results demonstrated that the RNAi plants had lower drought tolerance, displaying seriously withered leaves, lower relative growth rate, lower root-shoot ratio, and lower root activity under drought stress compared to WT plants. Physiological studies indicated that the suppression of CsGPA1 weakened drought stress tolerance due to higher water loss rate in the leaves, higher levels of hydrogen peroxide (H2O2), increased malondialdehyde (MDA) content, lower free proline content, lower soluble sugar content, lower soluble protein content, and decreased antioxidant enzyme activities. qRT-PCR analysis demonstrated that the interference of CsGPA1 up-regulated the expression of most AQP genes (except for CsPIP2;3 in leaves) and down-regulated the expression of CsPIP1;2, CsPIP1;4, CsPIP2;1, and CsPIP2;4 in roots under drought stress when compared to WT plants. Our results demonstrated that CsGPA1 could function as a positive regulator in drought stress response by decreasing the accumulation of reactive oxygen species (ROS), improving permeable potentials, and reducing water loss in cucumber plants.

Function analysis of ZmNAC33, a positive regulator in drought stress response in Arabidopsis

Drought significantly affects plant growth and has devastating effects on crop production, NAC transcription factors respond to abiotic stresses by activating gene expression. In this study, a maize NAC transcription factor, ZmNAC33, was cloned and characterized its function in Arabidopsis. Transient transformation in Arabidopsis leaves mesophyll protoplasts and trans-activation assays in yeast showed that ZmNAC33 was localized in the nucleus and had transactivation activity. qRT-PCR analysis showed that ZmNAC33 in maize was induced by drought, high salinity and abscisic acid (ABA) stress. Promoter analysis identified multiple stress-related cis-acting elements in the promoter region of ZmNAC33. In ZmNAC33 transgenic Arabidopsis, germination rates were higher than in wild type plants under ABA and osmotic stress at the germination stage, and overexpression lines exhibited higher survival rates and higher antioxidant enzyme activities compared with wild type under drought stress. These results indicate that ZmNAC33 actes as a positive regulator in drought tolerance in plants.

The Kinase CIPK11 Functions as a Negative Regulator in Drought Stress Response in Arabidopsis.

Drought is a major limiting factor for plant growth and crop productivity. Many Calcineurin B-like interacting protein kinases (CIPKs) play crucial roles in plant adaptation to environmental stresses. It is particularly essential to find the phosphorylation targets of CIPKs and to study the underlying molecular mechanisms. In this study, we demonstrate that CIPK11 acts as a novel component to modulate drought stress in plants. The overexpression of CIPK11 (CIPK11OE) in Arabidopsis resulted in the decreased tolerance of plant to drought stress. When compared to wild type plants, CIPK11OE plants exhibited higher leaf water loss and higher content of reactive oxygen species (ROS) after drought treatment. Additionally, a yeast two hybrid screening assay by using CIPK11 as a bait captures Di19-3, a Cys2/His2-type zinc-finger transcription factor that is involved in drought stress, as a new interactor of CIPK11. Biochemical analysis revealed that CIPK11 interacted with Di19-3 in vivo and it was capable of phosphorylating Di19-3 in vitro. Genetic studies revealed that the function of CIPK11 in regulating drought stress was dependent on Di19-3. The transcripts of stress responsive genes, such as RAB18, RD29A, RD29B, and DREB2A were down-regulated in the CIPK11OE plants. Whereas overexpression of CIPK11 in di19-3 mutant background, expression levels of those marker genes were not significantly altered. Taken together, our results demonstrate that CIPK11 partly mediates the drought stress response by regulating the transcription factor Di19-3.

Arabidopsis group XIV ubiquitin-conjugating enzymes AtUBC32, AtUBC33, and AtUBC34 play negative roles in drought stress response

AtUBC32, AtUBC33, and AtUBC34 comprise Arabidopsis group XIV E2 ubiquitin-conjugating enzymes. Yeast two-hybrid, in vitro pull-down, and bimolecular fluorescence complementation assays revealed that group XIV E2s are interacting partners of the U-box-type E3 ligase PUB19, a negative regulator of drought stress response. These three AtUBCs are co-localized with PUB19 to the punctae-like structures, most of which reside on the endoplasmic reticulum membrane of tobacco leaf cells. Suppression of AtUBC32, AtUBC33, and AtUBC34 resulted in increased abscisic acid-mediated stomatal closure and tolerance to drought stress. These results indicate that Arabidopsis group XIV E2s play negative roles in drought stress response.

Roles of a maize phytochrome-interacting factors protein ZmPIF3 in regulation of drought stress responses by controlling stomatal closure in transgenic rice without yield penalty.

ZmPIF3 plays an important role in ABA-mediated regulation of stomatal closure in the control of water loss, and can improve both drought tolerance and did not affect the grain yield in the transgenic rice. Phytochrome-interacting factors (PIFs) are a subfamily of basic helix-loop-helix (bHLH) transcription factors and play important roles in regulating plant growth and development. In our previous study, overexpression of a maize PIFs family gene, ZmPIF3, improved drought tolerance in transgenic rice. In this study, measurement of water loss rate, transpiration rate, stomatal conductance, guard cell aperture, density and length of ZmPIF3 transgenic plants showed that ZmPIF3 can enhance water-saving and drought-resistance by decreasing stomatal aperture and reducing transpiration in both transgenic rice and transgenic Arabidopsis. Scrutiny of sensitivity to ABA showed that ZmPIF3 transgenic rice was hypersensitive to ABA, while the endogenous ABA level was not significantly changed. These results indicate that ZmPIF3 plays a major role in the ABA signaling pathway. In addition, DGE results further suggest that ZmPIF3 participates in the ABA signaling pathway and regulates stomatal aperture in rice. Comparison analysis of the phenotype, physiology, and transcriptome of ZmPIF3 transgenic rice compared to control plants further suggests that ZmPIF3 is a positive regulator of ABA signaling and enhances water-saving and drought-resistance traits by reducing stomatal openings to control water loss. Moreover, investigation of the agronomic traits of ZmPIF3 transgenic rice from four cultivating seasons showed that ZmPIF3 expression increased the tiller and panicle number and did not affect the grain yield in the transgenic rice. These results demonstrate that ZmPIF3 is a promising candidate gene in the transgenic breeding of water-saving and drought-resistant rice plants and crop improvement.

Expression of a populus histone deacetylase gene 84KHDA903 in tobacco enhances drought tolerance

Histone deacetylases (HDACs) play a key role in regulating plant growth, development and stress responses. However, functions of HDACs in woody plants are largely unknown. In this study, a novel gene encoding a RPD3/HDA1-type histone deacetylase was cloned from 84K poplar (Populus alba×Populus glandulosa) and designated as 84KHDA903. The 84KHDA903 encodes a protein composed of 500 amino acid residues, which contains a conserved HDAC domain. Transient expression of 84KHDA903 in onion epidermal cells suggested that it was exclusively localized in nucleus. The 84KHDA903 exhibited different expression patterns under drought, salt and ABA treatments. The expression of 84KHDA903 was responsive to drought and ABA but not to salt. To understand the function of 84KHDA903 in stress responses, the 84KHDA903 gene was transformed into tobacco. The expression of 84KHDA903 in tobacco increased the tolerance of transgenic seeds to mannitol but not to salt. In adult stage, the 84KHDA903-expressing tobacco exhibited drought tolerance and showed strong capacity to recover after drought. During the recovery period, the stress-responsive genes including NtDREB4, NtDREB3 and NtLEA5 were induced to be highly expressed in the 84KHDA903 transgenic plants in contrast to wild-type plants. Taken together, for the first time, we reported a RPD3/HDA1-type histone deacetylase from poplar, 84KHDA903, which acted as a positive regulator in drought stress responses.

Genome-Wide Differences in DNA Methylation Changes in Two Contrasting Rice Genotypes in Response to Drought Conditions.

Differences in drought stress tolerance within diverse rice genotypes have been attributed to genetic diversity and epigenetic alterations. DNA methylation is an important epigenetic modification that influences diverse biological processes, but its effects on rice drought stress tolerance are poorly understood. In this study, methylated DNA immunoprecipitation sequencing and an Affymetrix GeneChip rice genome array were used to profile the DNA methylation patterns and transcriptomes of the drought-tolerant introgression line DK151 and its drought-sensitive recurrent parent IR64 under drought and control conditions. The introgression of donor genomic DNA induced genome-wide DNA methylation changes in DK151 plants. A total of 1190 differentially methylated regions (DMRs) were detected between the two genotypes under normal growth conditions, and the DMR-associated genes in DK151 plants were mainly related to stress response, programmed cell death, and nutrient reservoir activity, which are implicated to constitutive drought stress tolerance. A comparison of the DNA methylation changes in the two genotypes under drought conditions indicated that DK151 plants have a more stable methylome, with only 92 drought-induced DMRs, than IR64 plants with 506 DMRs. Gene ontology analyses of the DMR-associated genes in drought-stressed plants revealed that changes to the DNA methylation status of genotype-specific genes are associated with the epigenetic regulation of drought stress responses. Transcriptome analysis further helped to identify a set of 12 and 23 DMR-associated genes that were differentially expressed in DK151 and IR64, respectively, under drought stress compared with respective controls. Correlation analysis indicated that DNA methylation has various effects on gene expression, implying that it affects gene expression directly or indirectly through diverse regulatory pathways. Our results indicate that drought-induced alterations to DNA methylation may influence an epigenetic mechanism that regulates the expression of unique genes responsible for drought stress tolerance.