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[求助] proteomic analysis of the plant–virus interaction i

Comparative proteomic analysis of the plant–virus interaction in resistant and susceptible ecotypes of maize infected with sugarcane mosaic virus

ABSTRACT
Sugarcanemosaic virus (SCMV) is an important viral pathogen and has caused serious losses in grain and forage yield. To identify candidate SCMV resistance proteins and to explore the molecular mechanisms involved in the plant–SCMV interaction, we conducted proteomic analyses of leaf samples fromresistant and susceptible ecotypes ofmaize infected with SCMV.
Proteins were analyzed by quantitative two-dimensional differential gel electrophoresis (2D-DIGE), and 93 protein spots showed statistically significant differences after virus inoculation. Functional categorization showed that SCMV-responsive proteins were mainly involved in energy andmetabolism, stress and defense responses, photosynthesis, and carbon fixation. The majority of the identified proteins were located in chloroplast and cytoplasm based on bioinformatic analysis. Among these identified proteins, 17 have not been identified previously as virus-responsive proteins, and 7 were new and did not have assigned functions.
Western blotting analyses confirmed the expression patterns of proteins of specific interest, and the genes encoding these proteins were further analyzed by real-time PCR. The results of this study showed overlapping and specific proteomic responses to SCMV infection between resistant and susceptiblemaize ecotypes. This study provides further insight into themolecular events during compatible and incompatible interactions between viruses and host plants.
Biological significance
Sugarcane mosaic virus (SCMV) is an important viral pathogen and has caused serious losses in grain and forage yield. However, little is known about host–SCMV interactions from the proteome perspective. This study analyzed proteomic changes in resistant and susceptible plants that are infected with SCMV using DIGE based proteomics. We identified 17 proteins that have not been identified previously as virus-responsive proteins, and 7 new proteins without assigned functions. These proteins are interesting candidates for future research, as they may be associated with new biological functions and play important roles in plant–virus interactions. Real-time RT-PCR analysis of genes encoding several proteins of interest provided indication on whether the changes in protein abundance were regulated at the mRNA level.
The results of this study showed overlapping and specific proteomic responses to SCMV infection between resistant and susceptible ecotypes. After inoculation, the proteins involved in energy and metabolism, stress and defense responses, photosynthesis and other four functional groups showed significant changes in both ecotypes, which suggested that SCMV infection influenced these physiological processes in both the resistant Siyi and the susceptible Mo17. However, the oxidative burst was more pronounced during incompatible plant–SCMV interactions, as compared to those defined as compatible. We also observed an increase of enzymes involved in glycolysis and gluconeogenesis pathways in the resistant maize ecotype Siyi,while decrease in the susceptible maize ecotype Mo17. In addition, there is a marked increase of guanine nucleotide-binding protein beta submit in the resistant Siyi, which suggests a possible involvement of G-protein associated pathways in the resistant responses ofmaize to SCMV. These observations may possibly reveal protein targets/markers that are useful in the design of future diagnosis or plant protection strategies and provide new insights into the molecular mechanismof plant–virus interactions.
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