Overexpression of a Mitogen-Activated Protein Kinase SlMAPK3 Positively Regulates Tomato Tolerance to Cadmium and Drought Stress.
ABSTRACT: Mitogen-activated protein kinases (MAPKs) activation is a common defense response of plants to a range of abiotic stressors. SlMPK3, a serine-threonine protein kinase, has been reported as an important member of protein kinase cascade that also functions on plant stress tolerance. In this study, we cloned SlMPK3 from tomato and studied its role in cadmium (Cd2+) and drought tolerance. The results showed that transcripts of SlMAPK3 differentially accumulated in various plant tissues and were remarkably induced by different abiotic stressors and exogenous hormone treatments. Overexpression of SlMAPK3 increased tolerance to Cd2+ and drought as reflected by an increased germination rate and improved seedling growth. Furthermore, transgenic plants overexpressing SlMAPK3 showed an increased leaf chlorophyll content, root biomass accumulation and root activity under Cd2+ stress. Chlorophyll fluorescence analysis revealed that transgenic plants demonstrated an increased photosynthetic activity as well as contents of chlorophyll, proline, and sugar under drought stress. Notably, cadmium- and drought-induced oxidative stress was substantially attenuated in SlMAPK3 overexpressing plants as evidenced by lower malondialdehyde and hydrogen peroxide accumulation, and increased activity and transcript abundance of enzymatic antioxidants under stress conditions compared to that of wild-type. Our findings provide solid evidence that overexpression of SlMAPK3 gene in tomato positively regulates tolerance to Cd2+ and drought stress, which may have strengthen the molecular understanding of SlMAPK3 gene to improve abiotic stress tolerance.
Project description:Drought and high salinity are two major abiotic stresses that significantly affect agricultural crop productivity worldwide. Annexins are a multigene family that plays an essential role in plant stress responses and various cellular processes. Here, the AnnSp2 gene was cloned from drought-resistant wild tomato (Solanum pennellii) and functionally characterized in cultivated tomato. AnnSp2 protein was localized in the nucleus and had higher expression in leave, flower and fruit. It was induced by several phytohormones and some abiotic stresses. Tomato plants overexpressing AnnSp2 had increased tolerance to drought and salt stress, as determined by analysis of various physiological parameters. AnnSp2-transgenic plants were less sensitive to ABA during the seed germination and seedling stages. However, under drought stress, the ABA content significantly increased in the AnnSp2-overexpressing plants, inducing stomatal closure and reducing water loss, which underlay the plants' enhanced stress tolerance. Furthermore, scavenging reactive oxygen species (ROS), higher total chlorophyll content, lower lipid peroxidation levels, increased peroxidase activities (including APX, CAT and SOD) and higher levels of proline were observed in AnnSp2-overexpressing plants. These results indicate that overexpression of AnnSp2 in transgenic tomato improves salt and drought tolerance through ABA synthesis and the elimination of ROS.
Project description:BACKGROUND:Basic region/leucine zipper (bZIP) transcription factors perform as crucial regulators in ABA-mediated stress response in plants. Nevertheless, the functions for most bZIP family members in tomato remain to be deciphered. RESULTS:Here we examined the functional characterization of SlbZIP1 under salt and drought stresses in tomato. Silencing of SlbZIP1 in tomato resulted in reduced expression of multiple ABA biosynthesis- and signal transduction-related genes in transgenic plants. In stress assays, SlbZIP1-RNAi transgenic plants exhibited reduced tolerance to salt and drought stresses compared with WT plants, as are evaluated by multiple physiological parameters associated with stress responses, such as decreased ABA, chlorophyll contents and CAT activity, and increased MDA content. In addition, RNA-seq analysis of transgenic plants revealed that the transcription levels of multiple genes encoding defense proteins related to responses to abiotic stress (e.g. endochitinase, peroxidases, and lipid transfer proteins) and biotic stress (e.g. pathogenesis-related proteins) were downregulated in SlbZIP1-RNAi plants, suggesting that SlbZIP1 plays a role in regulating the genes related to biotic and abiotic stress response. CONCLUSIONS:Collectively, the data suggest that SlbZIP1 exerts an essential role in salt and drought stress tolerance through modulating an ABA-mediated pathway, and SlbZIP1 may hold potential applications in the engineering of salt- and drought-tolerant tomato cultivars.
Project description:Dehydrins are late embryogenesis abundant proteins that help regulate abiotic stress responses in plants. Overexpression of the Saussurea involucrata dehydrin gene SiDHN has previously been shown to improve water-use efficiency and enhance cold and drought tolerance of transgenic tobacco. To understand the mechanism by which SiDHN exerts its protective function, we transformed the SiDHN gene into tomato plants (Solanum lycopersicum L.) and assessed their response to abiotic stress. We observed that in response to stresses, the SiDHN transgenic tomato plants had increased contents of chlorophyll a and b, carotenoid and relative water content compared with wild-type plants. They also had higher maximal photochemical efficiency of photosystem II and accumulated more proline and soluble sugar. Compared to those wild-type plants, malondialdehyde content and relative electron leakage in transgenic plants were not significantly increased, and H2O2 and O2- contents in transgenic tomato plants were significantly decreased. We further observed that the production of stress-related antioxidant enzymes, including superoxide dismutase, ascorbate peroxidase, peroxidase, and catalase, as well as pyrroline-5-carboxylate synthetase and lipid transfer protein 1, were up-regulated in the transgenic plants under cold and drought stress. Based on these observations, we conclude that overexpression of SiDHN gene can promote cold and drought tolerance of transgenic tomato plants by inhibiting cell membrane damage, protecting chloroplasts, and enhancing the reactive oxygen species scavenging capacity. The finding can be beneficial for the application of SiDHN gene in improving crop tolerance to abiotic stress and oxidative damage.
Project description:Universal stress protein (USP) appears to play an active role in the abiotic stress response, but their functions remain largely unknown in plants. A USP gene (SpUSP) was cloned from wild tomato (Solanum pennellii) and functionally characterized in cultivated tomato in the present study. The SpUSP transcript is abundantly accumulated in leaf stomata and its expression varied with the circadian rhythm. SpUSP was remarkably induced by dehydration, salt stress, oxidative stress, and the phytohormone abscisic acid (ABA) etc. This protein was predominantly localized in the nucleus and cell membrane. Overexpressing SpUSP increased drought tolerance of tomato in the seedling and adult stages. Under drought stress, the ABA content significantly increased in the SpUSP-overexpressing plants, which induced stomatal closure and reduced water loss, leading to the enhancement of drought tolerance. Based on the microarray data, a large number of chlorophyll a/b-binding proteins and photosystem-related genes were up-regulated in the SpUSP-overexpressing plants under drought conditions, which possibly enhanced the stomatal sensivitity to ABA and maintained the photosynthetic function. SpUSP overexpression also alleviated the oxidative damage accompanied by oxidative stress-responsive gene activation and osmolyte accumulation. Annexin (SGN-U314161) was found to interacte with SpUSP in the yeast two-hybrid method. This interaction was further confirmed by the bimolecular fluorescence complementation assay. The present study demonstrated that the annexin-interacting SpUSP plays important roles in the drought tolerance of tomato by influencing ABA-induced stomatal movement, increasing photosynthesis, and alleviating oxidative stress.
Project description:The DEAD-box RNA helicases are involved in almost every aspect of RNA metabolism, associated with diverse cellular functions including plant growth and development, and their importance in response to biotic and abiotic stresses is only beginning to emerge. However, none of DEAD-box genes was well characterized in tomato so far. In this study, we reported on the identification and characterization of two putative DEAD-box RNA helicase genes, SlDEAD30 and SlDEAD31 from tomato, which were classified into stress-related DEAD-box proteins by phylogenetic analysis. Expression analysis indicated that SlDEAD30 was highly expressed in roots and mature leaves, while SlDEAD31 was constantly expressed in various tissues. Furthermore, the expression of both genes was induced mainly in roots under NaCl stress, and SlDEAD31 mRNA was also increased by heat, cold, and dehydration. In stress assays, transgenic tomato plants overexpressing SlDEAD31 exhibited dramatically enhanced salt tolerance and slightly improved drought resistance, which were simultaneously demonstrated by significantly enhanced expression of multiple biotic and abiotic stress-related genes, higher survival rate, relative water content (RWC) and chlorophyll content, and lower water loss rate and malondialdehyde (MDA) production compared to wild-type plants. Collectively, these results provide a preliminary characterization of SlDEAD30 and SlDEAD31 genes in tomato, and suggest that stress-responsive SlDEAD31 is essential for salt and drought tolerance and stress-related gene regulation in plants.
Project description:Abiotic stress induces nitrate (NO3 -) allocation to roots, which increases stress tolerance in plants. NRT1.1 is broadly involved in abiotic stress tolerance in plants, but the relationship between NRT1.1 and NO3 - allocation under stress conditions is unclear. In this study, we found that Arabidopsis wild-type Col-0 was more cadmium (Cd2+)-tolerant than the nrt1.1 mutant at 20 ?M CdCl2. Cd2+ exposure repressed NRT1.5 but upregulated NRT1.8 in roots of Col-0 plants, resulting in increased NO3 - allocation to roots and higher [NO3 -] root-to-shoot (R:S) ratios. Interestingly, NITRATE REGULATORY GENE2 (NRG2) was upregulated by Cd2+ stress in Col-0 but not in nrt1.1. Under Cd2+ stress, nrg2 and nrg2-3chl1-13 mutants exhibited similar phenotypes and NO3 - allocation patterns as observed in the nrt1.1 mutant, but overexpression of NRG2 in Col-0 and nrt1.1 increased the [NO3 -] R:S ratio and restored Cd2+ stress tolerance. Our results indicated that NRT1.1 and NRG2 regulated Cd2+ stress-induced NO3 - allocation to roots and that NRG2 functioned downstream of NRT1.1. Cd2+ uptake did not differ between Col-0 and nrt1.1, but Cd2+ allocation to roots was higher in Col-0 than in nrt1.1. Stressed Col-0 plants increased Cd2+ and NO3 - allocation to root vacuoles, which reduced their cytosolic allocation and transport to the shoots. Our results suggest that NRT1.1 regulates NO3 - allocation to roots by coordinating Cd2+ accumulation in root vacuoles, which facilitates Cd2+ detoxification.
Project description:Tomato (Solanum lycopersicum) is one of the most important vegetable crops; its production, productivity and quality are adversely affected by abiotic stresses. Abiotic stresses such as drought, extreme temperature and high salinity affect almost every stage of tomato life cycle. Depending upon the plant stage and duration of the stress, abiotic stress causes about 70% yield loss. Several wild tomato species have the stress tolerance genes; however, it is very difficult to transfer them into cultivars due to high genetic distance and crossing barriers. Transgenic technology is an alternative potential tool for the improvement of tomato crop to cope with abiotic stress, as it allows gene transfer across species. In recent decades, many transgenic tomatoes have been developed, and many more are under progress against abiotic stress using transgenes such as DREBs, Osmotin, ZAT12 and BADH2. The altered expression of these transgenes under abiotic stresses are involved in every step of stress responses, such as signaling, control of transcription, proteins and membrane protection, compatible solute (betaines, sugars, polyols, and amino acids) synthesis, and free-radical and toxic-compound scavenging. The stress-tolerant transgenic tomato development is based on introgression of a gene with known function in stress response and putative tolerance. Transgenic tomato plants have been developed against drought, heat and salt stress with the help of various transgenes, expression of which manages the stress at the cellular level by modulating the expression of downstream genes to ultimately improve growth and yield of tomato plants and help in sustainable agricultural production. The transgenic technology could be a faster way towards tomato improvement against abiotic stress. This review provides comprehensive information about transgenic tomato development against abiotic stress such as drought, heat and salinity for researcher attention and a better understanding of transgenic technology used in tomato improvement and sustainable agricultural production.
Project description:Salinity threatens productivity of economically important crops such as tomato (Solanum lycopersicum L.). WRKY transcription factors appear, from a growing body of knowledge, as important regulators of abiotic stresses tolerance. Tomato SlWRKY3 is a nuclear protein binding to the consensus CGTTGACC/T W box. SlWRKY3 is preferentially expressed in aged organs, and is rapidly induced by NaCl, KCl, and drought. In addition, SlWRKY3 responds to salicylic acid, and 35S::SlWRKY3 tomatoes showed under salt treatment reduced contents of salicylic acid. In tomato, overexpression of SlWRKY3 impacted multiple aspects of salinity tolerance. Indeed, salinized (125 mM NaCl, 20 days) 35S::SlWRKY3 tomato plants displayed reduced oxidative stress and proline contents compared to WT. Physiological parameters related to plant growth (shoot and root biomass) and photosynthesis (stomatal conductance and chlorophyll a content) were retained in transgenic plants, together with lower Na+ contents in leaves, and higher accumulation of K+ and Ca2+. Microarray analysis confirmed that many stress-related genes were already up-regulated in transgenic tomatoes under optimal conditions of growth, including genes coding for antioxidant enzymes, ion and water transporters, or plant defense proteins. Together, these results indicate that SlWRKY3 is an important regulator of salinity tolerance in tomato.
Project description:Plants can perceive environmental changes and respond to external stressors. Here, we show that OsNAC2, a member of the NAC transcription factor family, was strongly induced by ABA and osmotic stressors such as drought and high salt. With reduced yields under drought conditions at the flowering stage, OsNAC2 overexpression lines had lower resistance to high salt and drought conditions. RNAi plants showed enhanced tolerance to high salinity and drought stress at both the vegetative and flowering stages. Furthermore, RNAi plants had improved yields after drought stress. A microarray assay indicated that many ABA-dependent stress-related genes were down-regulated in OsNAC2 overexpression lines. We further confirmed that OsNAC2 directly binds the promoters of LATE EMBRYOGENESIS ABUNDANT 3 (OsLEA3) and Stress-Activated Protein Kinases 1 (OsSAPK1), two marker genes in the abiotic stress and ABA response pathways, respectively. Our results suggest that in rice OsNAC2 regulates both abiotic stress responses and ABA-mediated responses, and acts at the junction between the ABA and abiotic stress pathways.
Project description:The atmospheric CO2 concentration (a[CO2]) is increasing at an unprecedented pace. Exogenous melatonin plays positive roles in the response of plants to abiotic stresses, including drought and cold. The effect of elevated CO2 concentration (e[CO2]) accompanied by exogenous melatonin on plants under drought and cold stresses remains unknown. Here, tomato plants were grown under a[CO2] and e[CO2], with half of the plants pre-treated with melatonin. The plants were subsequently treated with drought stress followed by cold stress. The results showed that a decreased net photosynthetic rate (PN) was aggravated by a prolonged water deficit. The PN was partially restored after recovery from drought but stayed low under a successive cold stress. Starch content was downregulated by drought but upregulated by cold. The e[CO2] enhanced PN of the plants under non-stressed conditions, and moderate drought and recovery but not severe drought. Stomatal conductance (gs) and the transpiration rate (E) was less inhibited by drought under e[CO2] than under a[CO2]. Tomato grown under e[CO2] had better leaf cooling than under a[CO2] when subjected to drought. Moreover, melatonin enhanced PN during recovery from drought and cold stress, and enhanced biomass accumulation in tomato under e[CO2]. The chlorophyll a content in plants treated with melatonin was higher than in non-treated plants under e[CO2] during cold stress. Our findings will improve the knowledge on plant responses to abiotic stresses in a future [CO2]-rich environment accompanied by exogenous melatonin.