Multiple Stress Responsive Genes Research Articles

Mutations in the Staphylococcus aureus Global Regulator CodY Confer Tolerance to an Interspecies Redox-Active Antimicrobial.

Bacteria often exist in multispecies communities where interactions among different species can modify individual fitness and behavior. Although many competitive interactions have been characterized, molecular adaptations that can counter this antagonism and preserve or increase fitness remain underexplored. Here, we characterize the adaptation of Staphylococcus aureus to pyocyanin, a redox-active interspecies antimicrobial produced by Pseudomonas aeruginosa, a co-infecting pathogen frequently isolated from wound and chronic lung infections with S. aureus. Using experimental evolution, we identified mutations in a conserved global transcriptional regulator, CodY, that confer tolerance to pyocyanin and thereby enhance survival of S. aureus. The transcriptional response of a pyocyanin tolerant CodY mutant to pyocyanin indicated a two-pronged defensive response compared to the wild type. Firstly, the CodY mutant strongly suppressed metabolism, by downregulating pathways associated with core metabolism, especially translation-associated genes, upon exposure to pyocyanin. Metabolic suppression via ATP depletion was sufficient to provide comparable protection against pyocyanin to the wild-type strain. Secondly, while both the wild-type and CodY mutant strains upregulated oxidative stress response pathways, the CodY mutant overexpressed multiple stress response genes compared to the wild type. We determined that catalase overexpression was critical to pyocyanin tolerance as its absence eliminated tolerance in the CodY mutant and overexpression of catalase was sufficient to impart tolerance to the wild-type strain. Together, these results suggest that both transcriptional responses likely contribute to pyocyanin tolerance in the CodY mutant. Our data thus provide new mechanistic insight into adaptation toward interbacterial antagonism via altered regulation that facilitates multifaceted protective cellular responses.

Streptomyces alleviate abiotic stress in plant by producing pteridic acids

Soil microbiota can confer fitness advantages to plants and increase crop resilience to drought and other abiotic stressors. However, there is little evidence on the mechanisms correlating a microbial trait with plant abiotic stress tolerance. Here, we report that Streptomyces effectively alleviate drought and salinity stress by producing spiroketal polyketide pteridic acid H (1) and its isomer F (2), both of which promote root growth in Arabidopsis at a concentration of 1.3 nM under abiotic stress. Transcriptomics profiles show increased expression of multiple stress responsive genes in Arabidopsis seedlings after pteridic acids treatment. We confirm in vivo a bifunctional biosynthetic gene cluster for pteridic acids and antimicrobial elaiophylin production. We propose it is mainly disseminated by vertical transmission and is geographically distributed in various environments. This discovery reveals a perspective for understanding plant-Streptomyces interactions and provides a promising approach for utilising beneficial Streptomyces and their secondary metabolites in agriculture to mitigate the detrimental effects of climate change.

Metabolic versatility of soil microbial communities below the rocks of the hyperarid Dalangtan Playa.

The hyperarid Dalangtan Playa in the western Qaidam Basin, northwestern China, is a unique terrestrial analog of Mars. Despite the polyextreme environments of this area, habitats below translucent rocks capable of environmental buffering could serve as refuges for microbial life. In this study, the hybrid assembly of Illumina short reads and Nanopore long reads recovered high-quality and high-continuity genomes, allowing for high-accuracy analysis and a deeper understanding of extremophiles in the sheltered soils of the Dalangtan Playa. Our findings reveal self-supporting and metabolically versatile sheltered soil communities adapted to a hyperarid and hypersaline playa, which provides implications for the search for life signals on Mars.

Screening and identification of multiple abiotic stress responsive candidate genes based on hybrid-sequencing in Vicia sativa

Common vetch is an important leguminous forage for both livestock fodder and green manure and has a tremendous latent capacity in a sustainable agroecosystem. In the present study, a comprehensive transcriptome analysis of the aboveground leaves and underground roots of common vetch under multiple abiotic stress treatments, including NaCl, drought, cold, and cold drought, was performed using hybrid-sequencing technology, i. e. single-molecule real-time sequencing technology (SMRT) and supplemented by next-generation sequencing (NGS) technology. A total of 485,038 reads of insert (ROIs) with a mean length of 2606 bp and 228,261 full-length nonchimeric (FLNC) reads were generated. After deduplication, 39,709 transcripts were generated. Of these transcripts, we identified 1059 alternative splicing (AS) events, 17,227 simple sequence repeats (SSRs), and 1647 putative transcription factors (TFs). Furthermore, 640 candidates long noncoding RNAs (lncRNAs) and 28,256 complete coding sequences (CDSs) were identified. In gene annotation analyses, a total of 38,826 transcripts (97.78%) were annotated in eight public databases. Finally, seven multiple abiotic stress-responsive candidate genes were obtained through gene expression, annotation information, and protein-protein interaction (PPI) networks. Our research not only enriched the structural information of FL transcripts in common vetch, but also provided useful information for exploring the molecular mechanism of multiple abiotic stress tolerance between aboveground and underground tissues in common vetch and related legumes.

β-Cyclocitral, a Master Regulator of Multiple Stress-Responsive Genes in Solanum lycopersicum L. Plants

β-cyclocitral (βCC), a major apocarotenoid of β-carotene, enhances plants’ defense against environmental stresses. However, the knowledge of βCC’s involvement in the complex stress-signaling network is limited. Here we demonstrate how βCC reprograms the transcriptional responses that enable Solanum lycopersicum L. (tomato) plants to endure a plethora of environmental stresses. Comparative transcriptome analysis of control and βCC-treated tomato plants was done by generating RNA sequences in the BGISEQ-500 platform. The trimmed sequences were mapped on the tomato reference genome that identifies 211 protein-coding differentially expressed genes. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analysis and their enrichment uncovered that only upregulated genes are attributed to the stress response. Moreover, 80% of the upregulated genes are functionally related to abiotic and biotic stresses. Co-functional analysis of stress-responsive genes revealed a network of 18 genes that code for heat shock proteins, transcription factors (TFs), and calcium-binding proteins. The upregulation of jasmonic acid (JA)-dependent TFs (MYC2, MYB44, ERFs) but not the JA biosynthetic genes is surprising. However, the upregulation of DREB3, an abscisic acid (ABA)-independent TF, validates the unaltered expression of ABA biosynthetic genes. We conclude that βCC treatment upregulates multiple stress-responsive genes without eliciting JA and ABA biosynthesis.

Functional annotation of hypothetical proteins involved in multiple stress response in Oryza sativa

Plants being sessile in nature face multiple stresses during its growth and development. Understanding the dynamic tolerance mechanism of plants to changing environment is the need of the hour and developing climate smart crops is still a nightmare. Functional characterization of relevant genes is a prerequisite when identifying candidates for crop improvement, in this study we identified 66 multiple stress responsive genes in rice that could serve as potential candidates for breeding for stress tolerance. The 15 hypothetical genes rendering multiple stress tolerance were functionally annotated using a rich set of tools ranging from sequence similarity search to fold recognition methods. It is predicted that the query protein might be a copper chaperone for superoxide dismutase. A novel approach to annotate hypothetical proteins with different remote homology search and structure-based threading approaches was designed. In addition to identified multiple stress responsive proteins in rice (Oryza sativa L.), our study also provides a robust toolkit to annotate other hypothetical proteins in plants.

Overexpression of ZmWRKY65 transcription factor from maize confers stress resistances in transgenic Arabidopsis

Plant-specific WRKY transcription factors play important roles in regulating the expression of defense-responsive genes against pathogen attack. A multiple stress-responsive WRKY gene, ZmWRKY65, was identified in maize by screening salicylic acid (SA)-induced de novo transcriptomic sequences. The ZmWRKY65 protein was localized in the nucleus of mesophyll protoplasts. The analysis of the ZmWRKY65 promoter sequence indicated that it contains several stress-related transcriptional regulatory elements. Many environmental factors affecting the transcription of ZmWRKY65 gene, such as drought, salinity, high temperature and low temperature stress. Moreover, the transcription of ZmWRKY65 gene was also affected by the induction of defense related plant hormones such as SA and exogenous ABA. The results of seed germination and stomatal aperture assays indicated that transgenic Arabidopsis plants exhibit enhanced sensitivity to ABA and high concentrations of SA. Overexpression of ZmWRKY65 improved tolerance to both pathogen attack and abiotic stress in transgenic Arabidopsis plants and activated several stress-related genes such as RD29A, ERD10, and STZ as well as pathogenesis-related (PR) genes such as PR1, PR2 and PR5; these genes are involved in resistance to abiotic and biotic stresses in Arabidopsis. Together, this evidence implies that the ZmWRKY65 gene is involved in multiple stress signal transduction pathways.

Molecular characterization of stress tolerance genes associated with D. indicus strain under extreme environment conditions.

Deinococcus indicus is a novel bacteria isolated from West Bengal, India known for its UV radiation and heavy metal tolerance. Since, this organism is reported from a region known for heavy metal contamination and earlier investigations demonstrated its radiation resistance, our study focused on the multiple stress responsive and DNA repair mechanisms. Though, most of the members of the genus Deinococcus are Gram positive cocci, D. indicus postures Gram negative rod shaped cells. Hence, the objectives were framed precisely to understand DNA repair pathway and stress responsive genes expression with a broader perspective. Based on available whole genome sequence of D. indicus, quantitative real time PCR (qPCR) was done to determine the expression pattern of multiple stress responsive genes upon various environmental extremities. Among them, UV responsive genes like UvrD and UvsE showed elevated expression when subjected to UV-C radiation at different time intervals. Similarly, when supplemented with arsenic and chromium, ArsR and ArsB exhibited considerably higher level of expression. While all the genes were subsequently analyzed in-silico, depicted that most of them were with N-glycosylation site, GPI anchor sites, N-terminal trans-membrane helix region besides putative signal peptides. Overall, this study opined the functional information onstress tolerance genes that aid to understand the DNA damage recovery mechanism towards elucidation of DNA repair pathways.

Genomic and biotechnological interventions in Prosopis cineraria: current status, challenges and opportunities

The present review illustrates an inclusive overview on the progress made in the area of genomics, population genetics, phylogenetics, non-GM biotechnology and genomic resource development in Prosopis cineraria and also suggests how genomic and biotechnological research will accelerate gene discovery, molecular breeding and crop improvement program in P. cineraria. In the era of global climate change, plant resources able to tolerate adverse stressful environments particularly drought, heat, cold and salt stresses have immense importance. The adaptation of plants towards different abiotic stresses depend on the coordination and regulation of multiple stress-responsive genes. The tree species vital to arid environments possess a gene pool representing useful characteristics related to abiotic stresses. Utilization and characterization of genetic resources of plants of arid regions can lead to an increase in our knowledge of the genes that are linked to abiotic stress related traits. Prosopis cineraria is an important tree of Indian Thar desert and is able to survive in adverse environmental conditions. Genomics and identification of genomic resources of this important tree species may be useful in determination of its adaptive evolution under adverse environmental conditions. Little progress has been made for this tree species in the area of population genetics, genomic and genetic resources development in the form of transcriptome sequencing, generation of ESTs against the heat and drought stresses, and identification of genomic SSRs. The feasibility of micropropagation as a non-GM biotechnology has also been demonstrated in P. cineraria. In this review, we also compared the genomic resource development in P. cineraria with other Prosopis species and some other model forest tree species like Populus, Eucalyptus, Quercus, Pinus, Picea, Castanea. However, genomic resource developed in P. cineraria is almost negligible comparatively to other model plants and still considered as less studied species. Although, many stress-responsive genes have been identified in a number of model plants, the native plant species grown naturally under harsh environment are rich source of novel abiotic stress-tolerant genes. More efforts are therefore needed to characterise this tree species using most modern genomic, genetic and biotechnological tools. More genomic research would accelerate gene discovery and molecular breeding in P. cineraria and increase its acceptability as classical tree species at global level for abiotic stress tolerance.

OsPM1 is a positive regulator of rice tolerance to drought stress but a negative regulator of rice tolerance to salt stress

ABSTRACT To understand stress response mechanisms, a multiple stress-responsive gene OsPM1 with a disordered region containing about 30 amino acid residues was screened by microarray. Promoter region analysis using PlantCARE showed this gene contains multiple cis-regulatory elements responding to abiotic stresses. Quantitative real-time PCR and promoter-GUS transgenic plant analysis verified that OsPM1 was induced by multiple abiotic stresses. OsPM1-GFP fusion protein expression showed that OsPM1 was localized at the cell membrane. The rice plants overexpressing OsPM1 exhibited hypersensitivity to salt stress but enhanced drought tolerance compared to the wild-type plants; the rice plants expressing antisense of OsPM1 were hyposensitive to salt stress but with weak drought tolerance compared to the wild-type plants. These results showed OsPM1 is a positive regulator of rice tolerance to drought, but it’s also a negative regulator of rice tolerance to salt.

SlHyPRP1 and DEA1, the multiple stress responsive eight-cysteine motif family genes of tomato (Solanum lycopersicum L.) are expressed tissue specifically, localize and interact at cytoplasm and plasma membrane in vivo.

Owing to rapid global climate change, the occurrence of multiple abiotic stresses is known to influence the outburst of biotic stress factors which affects crop productivity. Therefore, it is essential to understand the molecular and cell biology of key genes associated with multiple stress responses in crop plants. SlHyPRP1 and DEA1, the members of eight-cysteine motif (8CM) family genes have been recently identified as putative regulators of multiple stress responses in tomato (Solanum lycopersicum L.). In order to gain deeper insight into cell and molecular biology of SlHyPRP1 and DEA1, we performed their expression analysis in three tomato cultivars and in vivo cell biological analysis. The semi-quantitative PCR and qRT-PCR results showed the higher expression of SlHyPRP1 and DEA1 in leaf, stem, flower and root tissues as compared to fruit and seed tissues in all three cultivars. The expression levels of SlHyPRP1 and DEA1 were found to be relatively higher in a wilt susceptible tomato cultivar (Arka Vikas) than a multiple disease resistant cultivar (Arka Abhed). In vivo cell biological analysis through Gateway cloning and Bi-FC assay revealed the predominant sub-cellular localization and strong protein-protein interaction of SlHyPRP1 and DEA1 at the cytoplasm and plasma membrane. Moreover, SlHyPRP1 showed in vivo interaction with stress responsive proteins WRKY3 and MST1. Our findings suggest that SlHyPRP1 with DEA1 are co-expressed with tissue specificity and might function together by association with WRKY3 and MST1 in plasma membrane for regulating multiple stress responses in the tomato plant.

Overexpression of Jatropha curcas ERFVII2 Transcription Factor Confers Low Oxygen Tolerance in Transgenic Arabidopsis by Modulating Expression of Metabolic Enzymes and Multiple Stress-Responsive Genes.

Enhancing crop tolerance to waterlogging is critical for improving food and biofuel security. In waterlogged soils, roots are exposed to a low oxygen environment. The group VII ethylene response factors (ERFVIIs) were recently identified as key regulators of plant low oxygen response. Oxygen-dependent N-end rule pathways can regulate the stability of ERFVIIs. This study aims to characterize the function of the Jatropha curcas ERFVIIs and the impact of N-terminal modification that stabilized the protein toward low oxygen response. This study revealed that all three JcERFVII proteins are substrates of the N-end rule pathway. Overexpression of JcERFVII2 conferred tolerance to low oxygen stress in Arabidopsis. In contrast, the constitutive overexpression of stabilized JcERFVII2 reduced low oxygen tolerance. RNA-seq was performed to elucidate the functional roles of JcERFVII2 and the impact of its N-terminal modification. Overexpression of both wildtype and stabilized JcERFVII2 constitutively upregulated the plant core hypoxia-responsive genes. Besides, overexpression of the stabilized JcERFVII2 further upregulated various genes controlling fermentative metabolic processes, oxidative stress, and pathogen responses under aerobic conditions. In summary, JcERFVII2 is an N-end rule regulated waterlogging-responsive transcription factor that modulates the expression of multiple stress-responsive genes; therefore, it is a potential candidate for molecular breeding of multiple stress-tolerant crops.

Brd4 inhibition ameliorates Pyocyanin-mediated macrophage dysfunction via transcriptional repression of reactive oxygen and nitrogen free radical pathways

Macrophages play critical roles in the first-line immune defense against airway infections caused by Pseudomonas aeruginosa (PA). The redox-active phenazine-pyocyanin (PCN), as one of the most essential virulence factors, facilities PA-related infection via a wide spectrum of cellular oxidative damages. However, little is known for PCN cytotoxicity in macrophages. In this study, besides showing PCN-mediated reactive oxygen species (ROS) indeed involved in macrophage viability and function impairment, we at the first time demonstrated a novel role of reactive nitrogen species (RNS) pathway causing cellular damage in PCN-challenged macrophages. Using small molecule inhibitor JQ1 targeting Bromodomain and extra-terminal family proteins, we showed restrained iNOS-dependent nitric oxide (NO) production correlated with abolished Brd4 recruitment to the NOS2 (encoding inducible nitric oxide synthase-iNOS) promoter. Application of JQ1 diminished PCN-mediated peroxynitrite (ONOO−) that followed ROS and NO induction, restored macrophage survival and bacteria clearance as well as repressed local inflammation in PA/PCN-challenged mice lungs. Our results uncover a novel link between PCN-mediated macrophage dysfunction and reactive free radicals that rely on Brd4-dependent transcription modulation of multiple stress-response genes, suggesting Brd4 could be a promising therapeutic target in treating PA-related lung infection.

WRKY genes family study reveals tissue-specific and stress-responsive TFs in wild potato species

Wild potatoes, as dynamic resource adapted to various environmental conditions, represent a powerful and informative reservoir of genes useful for breeding efforts. WRKY transcription factors (TFs) are encoded by one of the largest families in plants and are involved in several biological processes such as growth and development, signal transduction, and plant defence against stress. In this study, 79 and 84 genes encoding putative WRKY TFs have been identified in two wild potato relatives, Solanum commersonii and S. chacoense. Phylogenetic analysis of WRKY proteins divided ScWRKYs and SchWRKYs into three Groups and seven subGroups. Structural and phylogenetic comparative analyses suggested an interspecific variability of WRKYs. Analysis of gene expression profiles in different tissues and under various stresses allowed to select ScWRKY045 as a good candidate in wounding-response, ScWRKY055 as a bacterial infection triggered WRKY and ScWRKY023 as a multiple stress-responsive WRKY gene. Those WRKYs were further studied through interactome analysis allowing the identification of potential co-expression relationships between ScWRKYs/SchWRKYs and genes of various pathways. Overall, this study enabled the discrimination of WRKY genes that could be considered as potential candidates in both breeding programs and functional studies.

Mutation of chloroplast CHLM contributes to down-regulation of multiple stress response genes in Arabidopsis

Mg-protoporphyrin IX methyltransferase (CHLM) plays a crucial role in chlorophyll biosynthesis and converts Mg-protoporphyrin IX to Mg-protoporphyrin IX monomethyl ester. Previous research has indicated that down-expression of CHLM causes a pale green phenotype and Mg-protoporphyrin IX accumulation in Arabidopsis thaliana plants. Here, we report that a novel missense mutation (G to A transition) in the CHLM gene, results in the Gly59 → Glu amino acid substitution in the corresponding protein of Arabidopsis chlm-4 mutant plants. In silico modelling studies suggested that this substitution may disrupt membrane association of CHLM. Functional studies showed that the mutant form of CHLM leads to the excessive accumulation of superoxide anion radicals (O2−), although elimination of O2− could alleviate the yellow phenotype observed in chlm-4. In addition, qRT-PCR and western blotting analysis demonstrated that the expression of O2− scavengers, FSD1 and CSD2, were repressed in chlm-4 plants. The accumulation of photosystem II proteins was also decreased due to the CHLM mutation. Furthermore, we found that chlm-4 plants exhibited hypersensitive salt-stress during seed germination and down-expression of the salt stress-responsive CBFs-family genes. These results suggest CHLM positively regulates the expression of multiple photosynthetic-related and stress-responsive nuclear genes.

Engineering Multiple Abiotic Stress Tolerance in Canola, Brassica napus.

Impacts of climate change like global warming, drought, flooding, and other extreme events are posing severe challenges to global crop production. Contribution of Brassica napus towards the oilseed industry makes it an essential component of international trade and agroeconomics. Consequences from increasing occurrences of multiple abiotic stresses on this crop are leading to agroeconomic losses making it vital to endow B. napus crop with an ability to survive and maintain yield when faced with simultaneous exposure to multiple abiotic stresses. For an improved understanding of the stress sensing machinery, there is a need for analyzing regulatory pathways of multiple stress-responsive genes and other regulatory elements such as non-coding RNAs. However, our understanding of these pathways and their interactions in B. napus is far from complete. This review outlines the current knowledge of stress-responsive genes and their role in imparting multiple stress tolerance in B. napus. Analysis of network cross-talk through omics data mining is now making it possible to unravel the underlying complexity required for stress sensing and signaling in plants. Novel biotechnological approaches such as transgene-free genome editing and utilization of nanoparticles as gene delivery tools are also discussed. These can contribute to providing solutions for developing climate change resilient B. napus varieties with reduced regulatory limitations. The potential ability of synthetic biology to engineer and modify networks through fine-tuning of stress regulatory elements for plant responses to stress adaption is also highlighted.

Comparative study of DAM, Dof, and WRKY gene families in fourteen species and their expression in Vitis vinifera.

Bud dormancy is one of the most important defensive mechanisms through which plants resist cold stress during harsh winter weather. DAM, Dof, and WRKY have been reported to be involved in many biological processes, including bud dormancy. In the present study, grapevine (Vitis vinifera) and other thirteen plants (six woody plants and seven herbaceous plants) were analyzed for the quantity, sequence structure, and evolution patterns of their DAM, Dof, and WRKY gene family members. Moreover, the expression of VvDAM, VvDof, and VvWRKY genes was also investigated. Thus, 51 DAM, 1,205 WRKY, and 489 Dof genes were isolated from selected genomes, while 5 DAM, 114 WRKY, and 50 Dof duplicate gene pairs were identified in 10 genomes. Moreover, WGD and segmental duplication events were associated with the majority of the expansions of Dof and WRKY gene families. The VvDAM, VvDof, and VvWRKY genes significantly differentially expressed throughout bud dormancy outnumbered those significantly differentially expressed throughout fruit development or under abiotic stresses. Interestingly, multiple stress responsive genes were identified, such as VvDAM (VIT_00s0313g00070), two VvDof genes (VIT_18s0001g11310 and VIT_02s0025g02250), and two VvWRKY genes (VIT_07s0031g01710 and VIT_11s0052g00450). These data provide candidate genes for molecular biology research investigating bud dormancy and responses to abiotic stresses (namely salt, drought, copper, and waterlogging).

OsMSR3, a Small Heat Shock Protein, Confers Enhanced Tolerance to Copper Stress in Arabidopsis thaliana.

Copper is a mineral element essential for the normal growth and development of plants; however, excessive levels can severely affect plant growth and development. Oryza sativa L. multiple stress-responsive gene 3 (OsMSR3) is a small, low-molecular-weight heat shock protein (HSP) gene. A previous study has shown that OsMSR3 expression improves the tolerance of Arabidopsis to cadmium stress. However, the role of OsMSR3 in the Cu stress response of plants remains unclear, and, thus, this study aimed to elucidate this phenomenon in Arabidopsis thaliana, to further understand the role of small HSPs (sHSPs) in heavy metal resistance in plants. Under Cu stress, transgenic A. thaliana expressing OsMSR3 showed higher tolerance to Cu, longer roots, higher survival rates, biomass, and relative water content, and accumulated more Cu, abscisic acid (ABA), hydrogen peroxide, chlorophyll, carotenoid, superoxide dismutase, and peroxidase than wild-type plants did. Moreover, OsMSR3 expression in A. thaliana increased the expression of antioxidant-related and ABA-responsive genes. Collectively, our findings suggest that OsMSR3 played an important role in regulating Cu tolerance in plants and improved their tolerance to Cu stress through enhanced activation of antioxidative defense mechanisms and positive regulation of ABA-responsive gene expression.

SpPKE1, a Multiple Stress-Responsive Gene Confers Salt Tolerance in Tomato and Tobacco.

Understanding the mechanism of abiotic-tolerance and producing germplasm of abiotic tolerance are important in plant research. Wild species often show more tolerance of environmental stress factors than their cultivated counterparts. Genes from wild species show potential abilities to improve abiotic resistance in cultivated species. Here, a tomato proline-, lysine-, and glutamic-rich type gene SpPKE1 was isolated from abiotic-resistant species (Solanum pennellii LA0716) for over-expression in tomato and tobacco for salt tolerance. The protein encoded by SpPKE1 was predominantly localized in the cytoplasm in tobacco. SpPKE1 and SlPKE1 (from cultivated species S. lycopersicum cv. M82) shared 89.7% similarity in amino acid sequences and their transcripts abundance in flowers and fruits was reduced by the imposition of drought or oxidative stress and the exogenous supply of abscisic acid. The DNA of the PKE1 promoter was highly methylated in fruit and leaf, and the methylation of the coding sequence in leaf was significantly higher than that in fruit at different development stages. The over-expression of SpPKE1 under the control of a CaMV (Cauliflower Mosaic Virus) 35S promoter in transgenic tomato and tobacco plants enhanced their tolerance to salt stress. PKE1 was downregulated by abiotic stresses but enhanced the plant’s salt stress tolerance. Therefore, this gene may be involved in post-transcriptional regulation and may be an important candidate for molecular breeding of salt-tolerant plants.

Heterologous Expression of the DREB Transcription Factor AhDREB in Populus tomentosa Carrière Confers Tolerance to Salt without Growth Reduction under Greenhouse Conditions

The DREB transcription factors regulate multiple stress response genes, and are therefore useful for molecular plant breeding. AhDREB, a stress-inducible gene, was isolated from Atriplex hortensis L. and introduced into Populus tomentosa Carrière under the control of the CaMV35S promoter. Under salt stress, the chlorophyll content and net photosynthetic rate were higher in transgenic lines than in the wild type (WT). Moreover, the rate of electrolyte penetration (REC) was lower in the transgenic lines. Additional analyses revealed that the AhDREB transgenic plants generally displayed lower malondialdehyde (MDA) activity but higher superoxide dismutase (SOD) and peroxidase (POD) activities and proline content than the WT under salt stress. RNA sequencing indicated that AhDREB could enhance tolerance to salt by activating various downstream genes in the transgenic plants. Furthermore, no growth inhibition was detected in transgenic plants expressing AhDREB driven by the constitutive CaMV35S promoter. The transcriptome showed 165 and 52 differentially expressed genes in transgenic plants under stress and non-stress conditions, respectively, among which no significant metabolic pathway was enriched and no unintended effects have yet been identified. Together, these results suggest that AhDREB may be a good candidate gene for increasing salt tolerance in transgenic poplar breeding.