Characterization and Proteomic Analysis of Novel Rice Lesion Mimic Mutant with Enhanced Disease Resistance

Abstract

Lesion mimic mutants (LMMs) are plants that spontaneously form lesions without pathogen infection or external stimulus and exhibit resistance to pathogens. Here, a rice LMM was created by ethyl methane sulfonate mutagenesis, named as hpil (hydrogen peroxide induced lesion). Diaminobenzidine and trypan blue staining showed that large amounts of H2O2 were produced and cell death was occurred at and around the parts of lesion mimic in the rice leaves. The phenotype of hpil is controlled by a single recessive gene, localized at a 2 Mb interval on chromosome 2. The data suggested that hpil is a novel LMM with enhanced bacterial and fungal disease resistance, and multiple pathogenesis-related proteins (PRs) were up-regulated. The proteomes of leaves at three positions (different degrees of lesion mimic severity) were characterized in hpil compared with its wild type plant. Differentially expressed proteins were detected by two dimensional difference gel electrophoresis and 274 proteins were identified by MALDI TOF/TOF™. These proteins were related to metabolic process, cellular process and response to stimulus, with mostly down-regulated in hpil leaves. Many of these proteins were related to the Calvin cycle, photosynthetic electron transport chain, glycolysis/gluconeogenesis and phosphonates pathways. Some resistance-related proteins including 14-3-3 proteins, OsPR10 and antioxidases such as peroxidase, superoxide dismutase and ascorbate peroxidase were up-regulated in leaves with lesion mimic. These results provide the foundation for cloning of the target gene and shed light on the mechanism involved in autoimmunity of rice.

Keywords

• lesion mimic mutant
• H2O2
• disease resistance
• pathogenesis-related protein
• resistance- related pathway