ABSTRACT
Mesembryanthemum crystallinum (common ice plant) is a halophyte species that 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. McHB7-overexpressing in ice plant leaves through Agrobacterium-mediated transformation led to 25 times more McHB7 transcripts than the non-transformed wild type (WT). After 500 mM NaCl treatment for 7 days, the activities of superoxide dismutase (SOD) and peroxidase (POD) and water content of the transgenic plants were higher than the WT, while malondialdehyde (MDA) was decreased in the transgenic plants. A total of 1082 and 1072 proteins were profiled by proteomics under control and salt treatment, respectively, with 22 and 11 proteins uniquely identified under control and salt stress, respectively. Among the 11 proteins, 7 were increased and 4 were decreased after salt treatment. Most of the proteins whose expression increased in the McHB7 overexpression (OE) ice plants under high salinity were involved in transport regulation, catalytic activities, biosynthesis of secondary metabolites, and response to stimulus. The results demonstrate that the McHB7 transcription factor plays a positive role in improving plant salt tolerance.
Subject(s)
Mesembryanthemum/metabolism , Plant Proteins/metabolism , Proteomics , Salt Tolerance/physiology , Amino Acid Sequence , Cell Nucleus/drug effects , Cell Nucleus/metabolism , Computational Biology , Gene Expression Regulation, Plant/drug effects , Gene Ontology , Mesembryanthemum/drug effects , Mesembryanthemum/genetics , Phylogeny , Plant Leaves/drug effects , Plant Leaves/metabolism , Plant Proteins/genetics , Plants, Genetically Modified , Protein Transport/drug effects , Salinity , Salt Tolerance/drug effects , Salt Tolerance/genetics , Sodium Chloride/pharmacology , Stress, Physiological/drug effects , Stress, Physiological/genetics , Subcellular Fractions/metabolism , Transcription Factors/metabolismABSTRACT
Systemic Acquired Resistance (SAR) improves immunity of plant systemic tissue after local exposure to a pathogen. Guard cells that form stomatal pores on leaf surfaces recognize bacterial pathogens via pattern recognition receptors, such as Flagellin Sensitive 2 (FLS2). However, how SAR affects stomatal immunity is not known. In this study, we aim to reveal molecular mechanisms underlying the guard cell response to SAR using multi-omics of proteins, metabolites and lipids. Arabidopsis plants previously exposed to pathogenic bacteria Pseudomonas syringae pv. tomato DC3000 (Pst) exhibit an altered stomatal response compared to control plants when they are later exposed to the bacteria. Reduced stomatal apertures of SAR primed plants lead to decreased number of bacteria in leaves. Multi-omics has revealed molecular components of SAR response specific to guard cells functions, including potential roles of reactive oxygen species (ROS) and fatty acid signaling. Our results show an increase in palmitic acid and its derivative in the primed guard cells. Palmitic acid may play a role as an activator of FLS2, which initiates stomatal immune response. Improved understanding of how SAR signals affect stomatal immunity can aid biotechnology and marker-based breeding of crops for enhanced disease resistance.
Subject(s)
Arabidopsis/immunology , Disease Resistance/immunology , Lipidomics , Metabolomics , Plant Diseases/immunology , Plant Stomata/metabolism , Proteome/metabolism , Pseudomonas syringae/growth & development , Arabidopsis/metabolism , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Chlorophyll/metabolism , Chromatography, Liquid , Fatty Acids/metabolism , Mass Spectrometry , Palmitic Acids/metabolism , Plant Diseases/microbiology , Plant Leaves/metabolism , Plant Leaves/microbiology , Plant Stomata/immunology , Plant Stomata/microbiology , Protein Kinases/genetics , Protein Kinases/metabolism , Pseudomonas syringae/immunology , Pseudomonas syringae/pathogenicity , Reactive Oxygen Species/metabolismABSTRACT
Drought is one of the most widespread stresses leading to retardation of plant growth and development. We examined proteome changes of an important oil seed crop, canola (Brassica napus L.), under drought stress over a 14-day period. Using iTRAQ LC-MS/MS, we identified 1976 proteins expressed during drought stress. Among them, 417 proteins showed significant changes in abundance, and 136, 244, 286, and 213 proteins were differentially expressed in the third, seventh, 10th, and 14th day of stress, respectively. Functional analysis indicated that the number of proteins associated with metabolism, protein folding and degradation, and signaling decreased, while those related to energy (photosynthesis), protein synthesis, and stress and defense increased in response to drought stress. The seventh and 10th-day profiles were similar to each other but with more post-translational modifications (PTMs) at day 10. Interestingly, 181 proteins underwent PTMs; 49 of them were differentially changed in drought-stressed plants, and 33 were observed at the 10th day. Comparison of protein expression changes with those of gene transcription showed a positive correlation in B. napus, although different patterns between transcripts and proteins were observed at each time point. Under drought stress, most protein abundance changes may be attributed to gene transcription, and PTMs clearly contribute to protein diversity and functions.
