Mutation Mechanisms And Proteomic Analysis Of Rice Lesion Mimic Mutant Spl5

Lesion mimic mutant broadly represent a phenotypical class of mutants that spontaneously display cell death. Many mutants exhibiting HR-like lesions in the absence of pathogens attack, Most of LMMs constitutively activate immune responses such as callose deposition, inducible expression of defense-related genes, production of reactive oxygen species (ROS), and accumulation of phytoalexins, conferring enhanced resistance to infection by pathogens. Rice lesion mimic mutant spotted leaf5(spl5) has been reported that the spl5mutant broadly enhanced resistance to rice blast and bacterial blight pathogens. We determinated the ROS content and its scavenging enzymes activity of differentially degree necrosis spots of spl5leaves and found that O2’and H2O2content during the formation of necrotic spots in spl5leaves were more than wild-type. The reactive oxygen species (ROS) significantly accumulate in spl5leaves, which is an early feature of HR called as oxygen burst; The activities of some ROS scavenging enzymes, SOD, POD, APX and CAT also change in spl5, and APX and POD activity were corresponding increased while O2-and H2O2content increased. However, the CAT activity declined and SOD activity only increased in the more necrotic of the site of spl5than wild-type activity, suggesting that the imbalance of ROS metabolism may induce HR in spl5. we use protein two-dimensional electrophoresis analyze spl5mutant and wild-type, and isolate more than2000protein spots. And identify14differentially expression protein spots by software analysis and MALDI-TOF-TOF mass spectrometry analysis. These protein spots were involving pre-mRNA splicing, amino-acid metabolism, photosynthesis, glycolysis, ROS metabolism, defense-related and others proteins. Gene expression analysis of14genes by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that the mRNA level was most correlated with the protein level. And ascorbate peroxidase APX7, glutathione S-transferase GSTF14, chinase Chia2a, and glyceraldehyde-3-phosphate dehydrogenase (G3PDH) may with related of spl5cellular necrosis. S-adenosylmethionine synthetase1(SAMS) and Chia2a may with the relevant of spl5resistance, and so on.Here, we analyzed the spl5mutant to clarify necrosis way of lesion mimic mutant spl5and better understanding the necrosis mechanisms of lesion mimic mutant.