Project description:Mesembryanthemum crystallinum (common ice plant) is one of the facultative halophyte plants, and it serves as a model for investigating the molecular mechanisms underlying its salt stress response and tolerance. Here we cloned one of homeobox transcription factor (TF) gene McHB7 from ice plant, which has 60% similarity with the Arabidopsis AtHB7. Overexpression of McHB7 in Arabidopsis (OE) showed that the plants had significantly elevated relative water content (RWC), chlorophyll content, superoxide dismutase (SOD) and peroxidase (POD) activities after salt stress treatment. Proteomics analysis identified 145 to be significantly changed in abundance, and 66 were exclusively increased in the OE plants compared to wild type (WT). After salt treatment, 979 and 959 metabolites were significantly increased and decreased in OE plants compared to the WT, respectively. The results demonstrated McHB7 can improve photosynthesis and increase the leaf chlorophyll content, and affect TCA cycle by regulating metabolites (e.g., pyruvate) and proteins (e.g., citrate synthase). Also, McHB7 modulates the expression of stress-related proteins (e.g., superoxide dismutase, dehydroascorbate reductase and pyrroline-5-carboxylate synthase B) to scavenge reactive oxygen species and enhance plant salt tolerance.
Project description:Mesembryanthemum crystallinum (common ice plant) is one of the most important halophyte plants for plant stress biology research. In this study, we established an efficient Agrobacterium-mediated transformation method in ice plant and confirmed that ice plant can sustain gene overexpression for four weeks. Expression of a salt-induced transcription factor McHB7 (OE) reached the highest level at seven days after infiltration. Under salt and drought stresses, the growth of OE was better than wild type (WT), and the activities of redox enzymes and chlorophyll contents were higher in OE than the WT. Using proteomics, 475, 510 and 378 proteins were identified to be significantly changed in the OE lines under control, salt, and drought conditions, respectively. Most increased proteins were involved in various processes including Calvin cycle, citric acid cycle, glycolysis, and antioxidant pathways. Some were found to participate in ABA biosynthesis or response. Metabolomics revealed that many metabolites and phytohormones in OE were involved in plant growth and development. Also, ABA was increased in OE lines under control, salt, and drought conditions. Yeast one-hybrid analysis showed that McHB7 can bind to ERD and ABA-related motifs. And protein-protein interaction analysis discovered the candidate proteins that were responsive to stresses and hormones (e.g., ABA).
Project description:Mesembryanthemum crystallinum (common ice plant) is a facultative halophyte species, which has adapted to extreme conditions. In this study, we cloned a McHB7 transcription factor gene from the ice plant. The expression of McHB7 was significantly induced by 500 mM NaCl and it reached the peak under salt treatment for 7 days. The McHB7 protein was targeted to the nucleus. Overexpression of McHB7 gene in ice plant leaves by Agrobacterium-mediated transformation led to 25 times more McHB7 transcripts than the non-transformed control wild type (WT). After 500 mM NaCl treatment for 7 days, SOD, POD activities and water content of the transgenic plants were significantly higher than WT, while MDA content was decreased in the transgenic plants. Proteomics results showed that a total of 1082 and 1072 proteins were profiled under control and salt treatment, respectively. 22 (2%) and 11 (1%) proteins were uniquely identified under control and salt stress conditions, respectively. Among these 11 proteins, 7 were increased and 4 were decreased. Most identified proteins involved in the processes of regulation of biological, transporter and catalytic activity, biosynthesis of secondary metabolties and response to stimulus were significantly increased in the McHB7 overexpression ice plants under high salinity. All the results demonstrate that the McHB7 transcription factor plays a positive role in improving plant salt tolerance.
Project description:Comprehensive gene expression analysis of salt responses in a halophyte Mesembryanthemum crystallinum, ice plant, by RNAseq methods.