Drought Tolerance In Arabidopsis Research Articles

PagTPS1 and PagTPS10, the trehalose-6-phosphate synthase genes, increase trehalose content and enhance drought tolerance

Trehalose-6-phosphate synthase (TPS) genes play an active role in the trehalose metabolism pathway that regulates the responses of plants to diverse stresses. However, the functional identification, comparison, and conservatism of TPS genes in the responses of woody plants, especially poplars, to drought stress remain unclear. Here, the trehalose content of 84K (Populus alba × P. glandulosa) poplars was down-regulated and PagTPS and PagTPP genes had diverse response patterns under drought stress. Physicochemical properties, expression patterns, and functions of PagTPS1 and PagTPS10, two class I members of TPS gene family, were identified and compared. Transgenic 84K poplars overexpressing PagTPS1 and PagTPS10 had significantly higher trehalose content with approximately 138% and 123%, respectively, and stronger drought tolerance compared to WT. PagTPS1 and PagTPS10 promoted the expression of TPPA genes and drought-responsive genes. Accordingly, poplars inhibiting PagTPS1 and PagTPS10 expression via RNA interference had lower trehalose content and drought tolerance. Simultaneously, overexpressing PagTPS1 and PagTPS10 improved the trehalose content and drought tolerance of Arabidopsis. Overall, we proposed a model of the effects of PagTPS1 and PagTPS10 as conservative regulators on the responses of plants to drought, which would provide new insights into the functional explorations of TPS genes in plants.

The transcription factor TabZIP156 acts as a positive regulator in response to drought tolerance in Arabidopsis and wheat (Triticum aestivum L.)

Drought stress strongly restricts the growth, development, and yield of wheat worldwide. Among the various transcription factors (TFs) involved in the wheat drought response, the specific functions of many basic leucine zipper (bZIP) TFs related to drought tolerance are still not well understood. In this study, we focused on the bZIP TF TabZIP156 in wheat. Our analysis showed that TabZIP156 was highly expressed in both roots and leaves, and it responded to drought and abscisic acid (ABA) stress. Through subcellular localization and transactivation assays, we confirmed that TabZIP156 was located to the nucleus and functioned as a transcriptional activator. Overexpression of TabZIP156 in Arabidopsis enhanced drought tolerance, as evidenced by higher germination rate, longer root length, lower water loss rate, reduced ion leakage, increased proline accumulation, decreased levels of H2O2, O2− and MDA, and improved activities of POD, SOD, and CAT enzymes. Additionally, the expression of drought- and antioxidant-related genes were significantly upregulated in TabZIP156 transgenic Arabidopsis under drought stress. However, silencing TabZIP156 in wheat led to decreased proline content, increased accumulation of H2O2, O2− and MDA, reduced activities of antioxidant enzymes, and downregulation of many drought- and antioxidant-related genes under drought stress. Furthermore, the dual-luciferase assay demonstrated that TabZIP156 could activate the expression of TaP5CS, TaDREB1A, and TaPOD by binding to their promoters. Taken together, this study highlights the significant role of TabZIP156 in drought stress and provides valuable insights for its potential application in breeding drought-resistant wheat.

The U1 small nuclear RNA enhances drought tolerance in Arabidopsis.

Alternative splicing (AS) is an important post-transcriptional regulatory mechanism that improves plant tolerance to drought stress by modulating gene expression and generating proteome diversity. The interaction between the 5' end of U1 small nuclear RNA (U1 snRNA) and the conserved 5' splice site of precursor messenger RNA (pre-mRNA) is pivotal for U1 snRNP involvement in AS. However, the roles of U1 snRNA in drought stress responses remain unclear. This study provides a comprehensive analysis of AtU1 snRNA in Arabidopsis (Arabidopsis thaliana), revealing its high conservation at the 5' end and a distinctive four-leaf clover structure. AtU1 snRNA is localized in the nucleus and expressed in various tissues, with prominent expression in young floral buds, flowers, and siliques. Overexpression of AtU1 snRNA confers enhanced abiotic stress tolerance, as evidenced in seedlings by longer seedling primary root length, increased fresh weight, and a higher greening rate compared to the wild type. Mature AtU1 snRNA overexpressor plants exhibit higher survival rates and lower water loss rates under drought stress, accompanied by a significant decrease in H2O2 and increase in proline. This study also provides evidence of altered expression levels of drought-related genes in AtU1 snRNA overexpressor or genome-edited lines, reinforcing the crucial role of AtU1 snRNA in drought stress responses. Furthermore, the overexpression of AtU1 snRNA influences the splicing of downstream target genes, with a notable impact on SPEECHLESS (SPCH), a gene associated with stomatal development, potentially explaining the observed decrease in stomatal aperture and density. These findings elucidate the critical role of U1 snRNA as an AS regulator in enhancing drought stress tolerance in plants, contributing to a deeper understanding of the AS pathway in drought tolerance and increasing awareness of the molecular network governing drought tolerance in plants.

TaMYB44-5A reduces drought tolerance by repressing transcription of TaRD22-3A in the abscisic acid signaling pathway.

TaMYB44-5A identified as a transcription factor negatively regulates drought tolerance in transgenic Arabidopsis. Drought can severely reduce yields throughout the wheat-growing season. Many studies have shown that R2R3-MYB transcription factors are involved in drought stress responses. In this study, the R2R3-MYB transcription factor MYB44-5A was identified in wheat (Triticum aestivum L.) and functionally analyzed. Three homologs of TaMYB44 were isolated, all of which localized to the nucleus. Overexpression of TaMYB44-5A reduced drought tolerance in Arabidopsis thaliana. Further analysis showed that TaMYB44-5A reduced the sensitivity of transgenic Arabidopsis to ABA. Genetic and transcriptional regulation analyses demonstrated that the expression levels of drought- and ABA-responsive genes were downregulated by TaMYB44-5A, and TaMYB44-5A directly bound to the MYB-binding site on the promoter to repress the transcription level of TaRD22-3A. Our results provide insights into a novel molecular pathway in which the R2R3-MYB transcription factor negatively regulates ABA signaling in response to drought stress.

Genome-Wide Identification of NAC Transcription Factors in Chimonanthus praecox and Transgene CpNAC30 Affects Salt and Drought Tolerance in Arabidopsis

Genome-Wide Identification of NAC Transcription Factors in Chimonanthus praecox and Transgene CpNAC30 Affects Salt and Drought Tolerance in Arabidopsis

Drought-induced circular RNAs in maize roots: Separating signal from noise.

Circular RNAs (circRNAs) play an important role in diverse biological processes; however, their origin and functions, especially in plants, remain largely unclear. Here, we used 2 maize (Zea mays) inbred lines, as well as 14 of their derivative recombination inbred lines with different drought sensitivity, to systematically characterize 8,790 circRNAs in maize roots under well-watered (WW) and water-stress (WS) conditions. We found that a diverse set of circRNAs expressed at significantly higher levels under WS. Enhanced expression of circRNAs was associated with longer flanking introns and an enrichment of long interspersed nuclear element retrotransposable elements. The epigenetic marks found at the back-splicing junctions of circRNA-producing genes were markedly different from canonical splicing, characterized by increased levels of H3K36me3/H3K4me1, as well as decreased levels of H3K9Ac/H3K27Ac. We found that genes expressing circRNAs are subject to relaxed selection. The significant enrichment of trait-associated sites along their genic regions suggested that genes giving rise to circRNAs were associated with plant survival rate under drought stress, implying that circRNAs play roles in plant drought responses. Furthermore, we found that overexpression of circMED16, one of the drought-responsive circRNAs, enhances drought tolerance in Arabidopsis (Arabidopsis thaliana). Our results provide a framework for understanding the intricate interplay of epigenetic modifications and how they contribute to the fine-tuning of circRNA expression under drought stress.

Wheat BREVIS RADIX (BRX) regulates organ size, stomatal density and enhances drought tolerance in Arabidopsis

BREVIS RADIX (BRX) is a small plant-specific and evolutionary conserved gene family with divergent yet partially redundant biological functions including root and shoot growth, stomatal development and tiller angle in plants. We characterized a BRX family gene from wheat (Triticum aestivum) by gain-of-function in Arabidopsis. Overexpression of TaBRXL2A resulted in longer primary roots with increased root meristem size and higher root growth under control and exogenous hormone treatments as compared to wild type (Col-0) plants. Overexpression lines also exhibited significant differences with the wild type such as increased rosette size, higher leaf number and leaf size. At reproductive stage, overexpression lines exhibited wider siliques and higher grain weight per plant. Under drought stress, overexpression lines exhibited enhanced drought tolerance in terms of higher chlorophyll retention and lower oxidative stress, thereby leading to significant recovery from drought stress. The analysis suggests that the inherent lower stomatal density in the leaves of overexpression lines and higher stomatal closure in response to ABA might contribute to lower water loss from the overexpression lines. Furthermore, TaBRXL2A protein showed membrane localization, presence of conserved residues at N-terminal for palmitoylation, and phosphosites in the linker region which are prescribed for its potential role in protophloem differentiation and stomatal lineage. Thus, we identified a TaBRX family gene which is involved in developmental pathways essential for plant growth, and also enhances drought tolerance in Arabidopsis.

SbMYC2 mediates jasmonic acid signaling to improve drought tolerance via directly activating SbGR1 in sorghum.

SbMYC2 functions as a key regulator under JA signaling in enhancing drought tolerance of sorghum through direct activating SbGR1. Drought stress is one of the major threats to crop yield. In response to drought stress, functions of basic helix-loop-helix (bHLH) transcription factors (TFs) have been reported in Arabidopsis and rice, but little is known for sorghum. Here, we characterized the function of SbMYC2, a bHLH TF in sorghum, and found that SbMYC2 responded most significantly to PEG-simulated drought stress and JA treatments. Overexpression of SbMYC2 significantly enhanced drought tolerance in Arabidopsis, rice and sorghum. In addition, it reduced reactive oxygen species (ROS) accumulation and increased chlorophyll content in sorghum leaves. While silencing SbMYC2 by virus-induced gene silencing (VIGS) resulted in compromised drought tolerance of sorghum seedlings. Moreover, SbMYC2 can directly activate the expression of GLUTATHIONE-DISULFIDE REDUCTASE gene SbGR1. SbGR1 silencing led to significantly weakened drought tolerance of sorghum, and higher ROS accumulation and lower chlorophyll content in sorghum leaves were detected. In addition, SbMYC2 can interact with SbJAZs, suppressors of JA signaling, and thus can mediate JA signaling to activate SbGR1, thereby regulating sorghum's tolerance to drought stress. Overall, our findings demonstrate that bHLH TF SbMYC2 plays an important role in sorghum's response to drought stress, thus providing one theoretical basis for genetic enhancement of sorghum and even rice.

Genome-wide identification of GTE family proteins in sugarcane (Saccharum spontaneum) reveals that SsGTEL3a confers drought tolerance

The bromodomain is a highly conserved protein domain that specifically binds to acetylated lysine residues in histones, thereby activating transcription of target genes. Although some progress in Global Transcription Factor Group E (GTE) has been achieved in numerous animals and a few plant species, no systematic analysis of GTE gene families has been reported yet in sugarcane. In our study, 37 GTE and GTE-Like (GTEL) genes were characterized in the Saccharum spontaneum. All SsGTE/SsGTEL members were heterogeneously located on all chromosomes of the sugarcane genome and divided into five groups. Transcriptome data showed that SsGTEL3a was expressed at significantly higher levels under drought stress in drought-resistant varieties than in drought-sensitive varieties. Moreover, the overexpression of SsGTEL3a significantly improved the drought tolerance in Arabidopsis through improving the scavenging ability of reactive oxygen species. Additionally, an interaction between ScFAR1 and SsGTEL3a was identified, with ScFAR1 showing a positive response to drought stress in bacterium. In summary, this systematic analysis of GTE gene family in sugarcane and functional research of SsGTEL3a broadened deeper insight into their evolutionary dynamics and functional properties and provided new candidate genes for drought-resistant molecular breeding of sugarcane.

MdbHLH160 is stabilized via reduced MdBT2-mediated degradation to promote MdSOD1 and MdDREB2A-like expression for apple drought tolerance.

Drought stress is a key environmental factor limiting the productivity, quality, and geographic distribution of crops worldwide. Abscisic acid (ABA) plays an important role in plant drought stress responses, but the molecular mechanisms remain unclear. Here, we report an ABA-responsive bHLH transcription factor, MdbHLH160, which promotes drought tolerance in Arabidopsis (Arabidopsis thaliana) and apple (Malus domestica). Under drought conditions, MdbHLH160 is directly bound to the MdSOD1 (superoxide dismutase 1) promoter and activated its transcription, thereby triggering reactive oxygen species (ROS) scavenging and enhancing apple drought tolerance. MdbHLH160 also promoted MdSOD1 enzyme activity and accumulation in the nucleus through direct protein interactions, thus inhibiting excessive nuclear ROS levels. Moreover, MdbHLH160 directly upregulated the expression of MdDREB2A-like, a DREB (dehydration-responsive element binding factor) family gene that promotes apple drought tolerance. Protein degradation and ubiquitination assays showed that drought and ABA treatment stabilized MdbHLH160. The BTB protein MdBT2 was identified as an MdbHLH160-interacting protein that promoted MdbHLH160 ubiquitination and degradation, and ABA treatment substantially inhibited this process. Overall, our findings provide insights into the molecular mechanisms of ABA-modulated drought tolerance at both the transcriptional and post-translational levels via the ABA-MdBT2-MdbHLH160-MdSOD1/MdDREB2A-like cascade.

A Novel Isoprene Synthase from the Monocot Tree Copernicia prunifera (Arecaceae) Confers Enhanced Drought Tolerance in Transgenic Arabidopsis.

The capacity to emit isoprene, among other stresses, protects plants from drought, but the molecular mechanisms underlying this trait are only partly understood. The Arecaceae (palms) constitute a very interesting model system to test the involvement of isoprene in enhancing drought tolerance, as their high isoprene emissions may have contributed to make them hyperdominant in neotropical dry forests, characterized by recurrent and extended periods of drought stress. In this study we isolated and functionally characterized a novel isoprene synthase, the gene responsible for isoprene biosynthesis, from Copernicia prunifera, a palm from seasonally dry tropical forests. When overexpressed in the non-emitter Arabidopsis thaliana, CprISPS conferred significant levels of isoprene emission, together with enhanced tolerance to water limitation throughout plant growth and development, from germination to maturity. CprISPS overexpressors displayed higher germination, cotyledon/leaf greening, water usage efficiency, and survival than WT Arabidopsis under various types of water limitation. This increased drought tolerance was accompanied by a marked transcriptional up-regulation of both ABA-dependent and ABA-independent key drought response genes. Taken together, these results demonstrate the capacity of CprISPS to enhance drought tolerance in Arabidopsis and suggest that isoprene emission could have evolved in Arecaceae as an adaptive mechanism against drought.

A novel Pinellia ternata catalase gene PtCAT2 regulates drought tolerance in Arabidopsis by modulating ROS balance.

Drought is one of the major abiotic stresses limiting agricultural production, particularly for shallow-rooted plants like Pinellia ternata. It damages plants via oxidative burst, but this effect could be mitigated by catalase (CAT). However, no studies have been reported on CAT homologs in P. ternata, a drought-sensitive plant species. In the present study, a novel CAT gene, PtCAT2, was functionally characterized via overexpression in Arabidopsis and analysis of cis-elements in its promoter. The isolated CAT gene was 1479 bp and encoded a protein containing 242 amino acids. The protein contains the CAT activity motif and the heme-binding site of a typical CAT, and the subcellular analysis indicated that the protein localizes at the cytoplasm and membrane. Moreover, the quantitative real-time reverse transcription PCR indicated that PtCAT2 is expressed ubiquitously in P. ternata and is strongly induced by drought stress and abscisic acid (ABA) signals. PtCAT2 overexpression enhanced the drought tolerance of Arabidopsis, as shown by the 30% increase in plant survival and a five-fold- increase in CAT activity. Moreover, PtCAT2-transgenic plants increased superoxide dismutase and peroxidase activities and reduced malondialdehyde, membrane leakage, and hydrogen peroxide (H2O2) (P<0.05). Furthermore, PtCAT2-transgenic plants showed higher tolerance to oxidative stress caused by exogenous H2O2 and retained higher chlorophyll and water contents than the WT. The mitochondria function was better maintained as presented by the higher oxygen consumption rate in transgenics under drought stress (P<0.05). The endogenous CATs and drought response-related genes were also upregulated in transgenic lines under drought stress, indicating that PtCAT2 confers drought stress tolerance by enhancing the H2O2 scavenging ability of plants to maintain their membrane integrity. These results improve our understanding of the drought response mechanisms and provide a potential breeding strategy for P. ternata genetic improvement.

A B-Box Transcription Factor CoBBX24 from Camellia oleifera Delays Leaf Senescence and Enhances Drought Tolerance in Arabidopsis

Plants face various biotic and abiotic stress factors during their growth and development, among which, drought is a serious adverse factor that affects yield and quality in agriculture and forestry. Several transcription factors are involved in regulating plant responses to drought stress. In this study, the B-box (BBX) transcription factor CoBBX24 was cloned from Camellia oleifera. This gene encodes a 241-amino-acid polypeptide containing two B-box domains at the N-terminus. A phylogenetic analysis revealed that CoBBX24 and CsBBX24 from Camellia sinensis are in the same branch, with their amino acid sequences being identical by 96.96%. CoBBX24 was localized to the nucleus and acted as a transcriptional activator. The overexpression of CoBBX24 in Arabidopsis heightened its drought tolerance along with a relatively high survival rate, and the rate of water loss in the OX-CoBBX24 lines was observably lower than that of the wild-type. Compared to the wild-type, the root lengths of the OX-CoBBX24 lines were significantly inhibited with abscisic acid. Leaf senescence was delayed in the OX-CoBBX24 lines treated with abscisic acid. The expression of genes related to leaf senescence and chlorophyll breakdown (e.g., SAG12, SAG29, NYC1, NYE1, and NYE2) was downregulated in the OX-CoBBX24 lines. This study indicated that CoBBX24 positively regulates the drought tolerance in Arabidopsis through delayed leaf senescence.

Abiotic stress treatment reveals expansin like A gene OfEXLA1 improving salt and drought tolerance of Osmanthus fragrans by responding to abscisic acid

Sweet osmanthus (Osmanthus fragrans) is a having general approval aromatic tree in China that is widely applied to landscaping and gardening. However, the evergreen tree adaptability is limited by many environmental stresses. Currently, limited information is available regarding the genetic analysis and functional identification of expansin genes in response to abiotic stress in sweet osmanthus. In this study, a total of 29 expansin genes were identified and divided into four groups by genome-wide analysis from the sweet osmanthus genome. Transcriptome and quantitative Real-time PCR analysis showed that the cell wall-localized protein expansin-like A (OfEXLA1) gene was significantly induced by salt and drought treatment. Histochemical GUS staining of transgenic Arabidopsis lines in which GUS activity was driven with the OfEXLA1 promoter, GUS activity was significantly induced by salt, drought, and exogenous abscisic acid (ABA). In yeast, we found OfEXLA1 overexpression significantly improved the population of cells compared with wild-type strains after NaCl and polyethylene glycol (PEG) treatment. Additionally, OfEXLA1 overexpression not only promoted plant growth, but also improved the salt and drought tolerance in Arabidopsis. To gain insight into the role of ABA signaling in the regulation of OfEXLA1 improving abiotic tolerance in sweet osmanthus, four differentially expressed ABA Insensitive 5 (ABI5)-like genes (OfABL4, OfABL5, OfABL7, and OfABL8) were identified from transcriptome, and dual-luciferase (dual-LUC) and yeast one hybrid (Y1H) assay showed that OfABL4 and OfABL5 might bind to OfEXLA1 promoter to accumulate the OfEXLA1 expression by responding to ABA signaling to improve abiotic tolerance in sweet osmanthus. These results provide the information for understanding the molecular functions of expansin-like A gene and molecular breeding of sweet osmanthus in future.

Arabidopsis CPK6 regulates drought tolerance under high nitrogen by the phosphorylation of NRT1.1.

Nitrogen is an essential macronutrient for plant growth and development, and its availability to some extent is regulated by drought stress. Calcium-dependent protein kinases (CPKs) are a unique family of Ca 2+ sensors with diverse functions in nitrogen uptake and drought tolerance signaling pathways. However, how CPKs involve in the crosstalk between drought stress and nitrogen transportation remains largely unknown. Here we identify the drought tolerant function of Arabidopsis CPK6 under high nitrogen condition. CPK6 expression is induced by the treatments of ABA and drought. The mutant cpk6 is insensitive to the ABA treatment and low nitrogen, but sensitive to drought only under high nitrogen condition. CPK6 interacts with NRT1.1 protein and phosphorylates the Thr447 residue, which then represses the NO3 - transporting activity of Arabidopsis under HN and drought stress. Taken together, we showed the evidences that CPK6 regulates Arabidopsis drought tolerance through changing NRT1.1 phosphorylation states, and that enriches the knowledge of nitrogen uptake in plants during drought stress.

Rhizobacteria isolated from xerophyte Haloxylon ammodendron manipulate root system architecture and enhance drought and salt tolerance in Arabidopsis thaliana.

The objective of this study was to identify bacteria from the rhizosphere of the black saxaul (Haloxylon ammodendron) and test the possibility of using the bacteria for enhancement of drought and/or salt tolerance in the model plant, Arabidopsis thaliana. We collected rhizosphere and bulk soil samples from a natural habitat of H. ammodendron in Iran and identified 58 morphotypes of bacteria that were enriched in the rhizosphere. From this collection, we focused our further experiments on eight isolates. Microbiological analyses showed that these isolates have different levels of tolerance to heat, salt, and drought stresses, and showed different capabilities of auxin production and phosphorous solubilization. We first tested the effects of these bacteria on the salt tolerance of Arabidopsis on agar plate assays. The bacteria substantially influenced the root system architecture, but they were not effective in increasing salt tolerance significantly. Pot assays were then conducted to evaluate the effects of the bacteria on salt or drought tolerance of Arabidopsis on peat moss. Results showed that three of these bacteria (Pseudomonas spp. and Peribacillus sp.) effectively enhanced drought tolerance in Arabidopsis, so that while none of the mock-inoculated plants survived after 19days of water withholding, the survival rate was 50-100% for the plants that were inoculated with these bacteria. The positive effects of the rhizobacteria on a phylogenetically-distant plant species imply that the desert rhizobacteria may be used to enhance abiotic stress in crops.

Long noncoding RNA ARTA controls ABA response through MYB7 nuclear trafficking in Arabidopsis.

In eukaryotes, transcription factors are a crucial element in the regulation of gene expression, and nuclear translocation is the key to the function of transcription factors. Here, we show that the long intergenic noncoding RNA ARTA interacts with an importin β-like protein, SAD2, through a long noncoding RNA-binding region embedded in the carboxyl terminal, and then it blocks the import of the transcription factor MYB7 into the nucleus. Abscisic acid (ABA)-induced ARTA expression can positively regulate ABI5 expression by fine-tuning MYB7 nuclear trafficking. Therefore, the mutation of arta represses ABI5 expression, resulting in desensitization to ABA, thereby reducing Arabidopsis drought tolerance. Our results demonstrate that lncRNA can hijack a nuclear trafficking receptor to modulate the nuclear import of a transcription factor during plant responses to environmental stimuli.

A Surprising Diversity of Xyloglucan Endotransglucosylase/Hydrolase in Wheat: New in Sight to the Roles in Drought Tolerance.

Drought has become a major limiting factor for wheat productivity, and its negative impact on crop growth is anticipated to increase with climate deterioration in arid areas. Xyloglucan endoglycosylases/hydrolases (XTHs) are involved in constructing and remodeling cell wall structures and play an essential role in regulating cell wall extensibility and stress responses. However, there are no systematic studies on the wheat XTH gene family. In this study, 71 wheat XTH genes (TaXTHs) were characterized and classified into three subgroups through phylogenetic analysis. Genomic replication promoted the expansion of TaXTHs. We found a catalytically active motif and a potential N-linked glycosylation domain in all TaXTHs. Further expression analysis revealed that many TaXTHs in the roots and shoots were significantly associated with drought stress. The wheat TaXTH12.5a gene was transferred into Arabidopsis to verify a possible role of TaXTHs in stress response. The transgenic plants possessed higher seed germination rates and longer roots and exhibited improved tolerance to drought. In conclusion, bioinformatics and gene expression pattern analysis indicated that the TaXTH genes played a role in regulating drought response in wheat. The expression of TaXTH12.5a enhanced drought tolerance in Arabidopsis and supported the XTH genes' role in regulating drought stress response in plants.

Dioscorea composita WRKY5 positively regulates AtSOD1 and AtABF2 to enhance drought and salt tolerances.

DcWRKY5 increases the antioxidant enzyme activity and proline accumulation, oppositely, reduces the accumulation of ROS and MDA, through directly activating the genes expression, finally enhances the salt and drought tolerance. Drought and salinity are two main environmental factors that limit the large-scale cultivation of the medicinal plant Dioscorea composita (D. composita). WRKY transcription factors (TFs) play vital roles in regulating drought and salt tolerance in plants. Nevertheless, the molecular mechanism of WRKY TF mediates drought and salt resistance of D. composita remains largely unknown. Here, we isolated and characterized a WRKY TF from D. composita, namely DcWRKY5, which was localized to the nucleus and bound to the W-box cis-acting elements. Expression pattern analysis showed that it was highly expressed in root and significantly up-regulated in the presence of salt, polyethylene glycol-6000 (PEG-6000) and abscisic acid (ABA). Heterologous expression of DcWRKY5 increased salt and drought tolerance in Arabidopsis, but was insensitive to ABA. In addition, compared with the wild type, the DcWRKY5 overexpressing transgenic lines had more proline, higher antioxidant enzyme (POD, SOD, and CAT) activities, less reactive oxygen species (ROS) and malondialdehyde (MDA). Correspondingly, the overexpression of DcWRKY5 modulated the expression of genes related to salt and drought stresses, such as AtSS1, AtP5CS1, AtCAT, AtSOD1, AtRD22, and AtABF2. Dual luciferase assay and Y1H were further confirmed that DcWRKY5 activate the promoter of AtSOD1 and AtABF2 through directly binding to the enrichment region of the W-box cis-acting elements. These results suggest that DcWRKY5 is a positive regulator of the drought and salt tolerance in D. composita and has potential applications in transgenic breeding.

Receptor-like cytoplasmic kinase ScRIPK in sugarcane regulates disease resistance and drought tolerance in Arabidopsis.

Receptor-like cytoplastic kinases (RLCKs) are known in many plants to be involved in various processes of plant growth and development and regulate plant immunity to pathogen infection. Environmental stimuli such as pathogen infection and drought restrict the crop yield and interfere with plant growth. However, the function of RLCKs in sugarcane remains unclear. In this study, a member of the RLCK VII subfamily, ScRIPK, was identified in sugarcane based on sequence similarity to the rice and Arabidopsis RLCKs. ScRIPK was localized to the plasma membrane, as predicted, and the expression of ScRIPK was responsive to polyethylene glycol treatment and Fusarium sacchari infection. Overexpression of ScRIPK in Arabidopsis enhanced drought tolerance and disease susceptibility of seedlings. Moreover, the crystal structure of the ScRIPK kinase domain (ScRIPK KD) and the mutant proteins (ScRIPK-KD K124R and ScRIPK-KD S253A|T254A) were characterized in order to determine the activation mechanism. We also identified ScRIN4 as the interacting protein of ScRIPK. Our work identified a RLCK in sugarcane, providing a potential target for sugarcane responses to disease infection and drought, and a structural basis for kinase activation mechanisms.