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Overexpression of the Maize Sulfite Oxidase Increases Sulfate and GSH Levels and Enhances Drought Tolerance in Transgenic Tobacco.

ABSTRACT: Sulfite oxidase (SO) plays a pivotal role in sulfite metabolism. In our previous study, sulfite-oxidizing function of the SO from Zea mays (ZmSO) was characterized. To date, the knowledge of ZmSO's involvement in abiotic stress response is scarce. In this study, we aimed to investigate the role of ZmSO in drought stress. The transcript levels of ZmSO were relatively high in leaves and immature embryos of maize plants, and were up-regulated markedly by PEG-induced water stress. Overexpression of ZmSO improved drought tolerance in tobacco. ZmSO-overexpressing transgenic plants showed higher sulfate and glutathione (GSH) levels but lower hydrogen peroxide (H2O2) and malondialdehyde (MDA) contents under drought stress, indicating that ZmSO confers drought tolerance by enhancing GSH-dependent antioxidant system that scavenged ROS and reduced membrane injury. In addition, the transgenic plants exhibited more increased stomatal response than the wild-type (WT) to water deficit. Interestingly, application of exogenous GSH effectively alleviated growth inhibition in both WT and transgenic plants under drought conditions. qPCR analysis revealed that the expression of several sulfur metabolism-related genes was significantly elevated in the ZmSO-overexpressing lines. Taken together, these results imply that ZmSO confers enhanced drought tolerance in transgenic tobacco plants possibly through affecting stomatal regulation, GSH-dependent antioxidant system, and sulfur metabolism-related gene expression. ZmSO could be exploited for developing drought-tolerant maize varieties in molecular breeding.

SUBMITTER: Xia Z

PROVIDER: S-EPMC5857591 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Drought adversely affects crop growth and yields. The cloning and characterization of drought- or abscisic acid (ABA)-inducible promoters is of great significance for their utilization in the genetic improvement of crop resistance. Our previous studies have shown that maize sulfite oxidase (SO) has a sulfite-oxidizing function and is involved in the drought stress response. However, the promoter of the maize SO gene has not yet been characterized. In this study, the promoter (ZmSOPro, 1194 bp upstream region of the translation initiation site) was isolated from the maize genome. The in-silico analysis of the ZmSOPro promoter identified several cis-elements responsive to the phytohormone ABA and drought stress such as ABA-responsive element (ABRE) and MYB binding site (MBS), besides a number of core cis-acting elements, such as TATA-box and CAAT-box. A 5' RACE (rapid amplification of cDNA ends) assay identified an adenine residue as the transcription start site of the ZmSO. The ZmSOPro activity was detected by β-glucuronidase (GUS) staining at nearly all developmental stages and in most plant organs, except for the roots in transgenic Arabidopsis. Moreover, its activity was significantly induced by ABA and drought stress. The 5'-deletion mutant analysis of the ZmSOPro in tobacco plants revealed that a 119-bp fragment in the ZmSOPro (upstream of the transcription start site) is a minimal region, which is required for its high-level expression. Moreover, the minimal ZmSOPro was significantly activated by ABA or drought stress in transgenic plants. Further mutant analysis indicated that the MBS element in the minimal ZmSOPro region (119 bp upstream of the transcription start site) is responsible for ABA and drought-stress induced expression. These results improve our understanding of the transcriptional regulation mechanism of the ZmSO gene, and the characterized 119-bp promoter fragment could be an ideal candidate for drought-tolerant gene engineering in both monocot and dicot crops.

Project description:Restriction in nutrient acquisition is one of the primary causes for reduced growth and yield in water deficient soils. Sulfur (S) is an important secondary macronutrient that interacts with several stress metabolites to improve performance of food crops under various environmental stresses including drought. Increased S supply influences uptake and distribution of essential nutrients to confer nutritional homeostasis in plants exposed to limited water conditions. The regulation of S metabolism in plants, resulting in synthesis of numerous S-containing compounds, is crucial to the acclimation response to drought stress. Two different experiments were laid out in semi-controlled conditions to investigate the effects of different S sources on physiological and biochemical mechanisms of maize (Zea mays L. cv. P1574). Initially, the rate of S application in maize was optimized in terms of improved biomass and nutrient uptake. The maize seedlings were grown in sandy loam soil fertigated with various doses (0, 15, 30 and 45?kg?ha-1) of different S fertilizers viz. K2SO4, FeSO4, CuSO4 and Na2SO4. The optimized S dose of each fertilizer was later tested in second experiment to determine its role in improving drought tolerance of maize plants. A marked effect of S fertilization was observed on biomass accumulation and nutrients uptake in maize. In addition, the optimized doses significantly increased the gas exchange characteristics and activity of antioxidant enzymes to improve yield of maize. Among various S sources, application of K2SO4 resulted in maximum photosynthetic rate (43%), stomatal conductance (98%), transpiration rate (61%) and sub-stomatal conductance (127%) compared to no S supply. Moreover, it also increased catalase, guaiacol peroxidase and superoxide dismutase activities by 55, 87 and 65%, respectively that ultimately improved maize yield by 33% with respect to control under water deficit conditions. These results highlight the importance of S fertilizers that would likely be helpful for farmers to get better yield in water deficient soils.

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Overexpression of the Maize Sulfite Oxidase Increases Sulfate and GSH Levels and Enhances Drought Tolerance in Transgenic Tobacco.

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