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Comparative phosphoproteomic analysis of blast resistant and susceptible rice cultivars in response to salicylic acid.

ABSTRACT: BACKGROUND:Salicylic acid (SA) is a significant signaling molecule that induces rice resistance against pathogen invasion. Protein phosphorylation carries out an important regulatory function in plant defense responses, while the global phosphoproteome changes in rice response to SA-mediated defense response has not been reported. In this study, a comparative phosphoproteomic profiling was conducted by two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) analysis, with two near-isogenic rice cultivars after SA treatment. RESULTS:Thirty-seven phosphoprotein spots were differentially expressed after SA treatment, twenty-nine of which were identified by MALDI-TOF/TOF MS, belonging to nine functional categories. Phosphoproteins involved in photosynthesis, antioxidative enzymes, molecular chaperones were similarly expressed in the two cultivars, suggesting SA might alleviate decreases in plant photosynthesis, regulate the antioxidant defense activities, thus improving basal resistance response in both cultivars. Meanwhile, phosphoproteins related to defense, carbohydrate metabolism, protein synthesis and degradation were differentially expressed, suggesting phosphorylation regulation mediated by SA may coordinate complex cellular activities in the two cultivars. Furthermore, the phosphorylation sites of four identified phosphoproteins were verified by NanoLC-MS/MS, and phosphorylated regulation of three enzymes (cinnamoyl-CoA reductase, phosphoglycerate mutase and ascorbate peroxidase) was validated by activity determination. CONCLUSIONS:Our study suggested that phosphorylation regulation mediated by SA may contribute to the different resistance response of the two cultivars. To our knowledge, this is the first report to measure rice phosphoproteomic changes in response to SA, which provides new insights into molecular mechanisms of SA-induced rice defense.

SUBMITTER: Sun R

PROVIDER: S-EPMC6819546 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Comparative phosphoproteomic analysis of blast resistant and susceptible rice cultivars in response to salicylic acid.

Sun Ranran R Qin Shiwen S Zhang Tong T Wang Zhenzhong Z Li Huaping H Li Yunfeng Y Nie Yanfang Y

BMC plant biology 20191028 1

<h4>Background</h4>Salicylic acid (SA) is a significant signaling molecule that induces rice resistance against pathogen invasion. Protein phosphorylation carries out an important regulatory function in plant defense responses, while the global phosphoproteome changes in rice response to SA-mediated defense response has not been reported. In this study, a comparative phosphoproteomic profiling was conducted by two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) analysis, with t ...[more]

PMID: 31660870

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Project description:BackgroundIn higher plants, salicylic acid (SA) plays important roles in inducing resistance to biotic and abiotic stresses. Tomato yellow leaf curl virus (TYLCV) causes a highly devastating viral disease in plants, particularly in tomato. However, the roles of SA in inducing tomato plant resistance to TYLCV remain unclear.ResultsIn this study, we investigated whether the exogenous application of SA can improve the resistance of tomato plants to TYLCV in two tomato cultivars, resistant 'Zhefen-702' and susceptible 'Jinpeng-1'. The impacts of SA on the accumulation of ascorbic acid (AsA) and biosynthetic gene expression, the activity of some important reactive oxygen species (ROS)-scavenging enzymes, and the expression patterns of stress-related genes were also determined. Results indicated that SA can effectively regulate the accumulation of AsA, especially in 'Jinpeng-1'. Similarly, the expression patterns of most of the AsA biosynthetic genes showed a negative relationship with AsA accumulation in the resistant and susceptible tomato cultivars. In the two tomato cultivars, the activities of ascorbate peroxidase (APX) and peroxidase (POD) in the SA + TYLCV treated plants were increased during the experiment period except at 14 days (APX in 'Jinpeng-1' was also at 4 days) post infected (dpi) with TYLCV. Simultaneously, the activity of SOD was reduced in 'Jinpeng-1' and increased in 'Zhefen-702' after treatment with SA + TYLCV. SA can substantially induce the expression of ROS-scavenging genes at different extents. From 2 to 10 dpi, the virus content in the SA + TYLCV treated plants was remarkably lower than those in the TYLCV treated plants in 'Jinpeng-1'and Zhefen-702'.ConclusionsThe above results suggest that SA can enhance tomato plant resistance by modulating the expression of genes encoding for ROS-scavenging players, altering the activity of resistance-related enzymes, and inducing the expression of pathogenesis-related genes to produce systemic acquired resistance. Simultaneously, these results confirm that SA is a resistance-inducing factor against TYLCV infection that can be effectively applied in tomato plants.

Project description:Purpose: To study the differential gene expression by comparing resistant and susceptible rice cultivars against the rice blast fungus M. Oryzae. Methods: Rice leaves were collected after M. Oryzae (MO36) infection from resistant and susceptible cultivars, preserved on deep freezer on -80O C frozen on liquid nitrogen, Total RNA from rice plant tissues was extracted using RNeasy Plant Mini Kit (Qiagen) by following manufacturer protocol. RNA libraries were prepared for sequencing using standard Illumina protocols. Results: Around 35 million pairs of filtered 150 base paired reads were obtained for each biological sample. These reads were mapped against RGAP7 using reference assembly tool of CLC Genomics Workbench. The mapped read data was summarised in to a matrix for each sample. count data was compared between the samples to identify the differentially expressed genes. genes were further filtered based on fold change, p value and FDR p value. The count data was further statistically analysed using Kal's Z test integrated into CGWB. Genes exhibiting 3 fold change and FDR p value <0.05 were filtered as either up regulated or down regulated gene. we obtained 7577 and 4290 significant (FDR adjusted p value ≤0.05) differentially expressed locus id in the resistant and susceptible rice cultivars, respectively . The number of significant DEL with fold change value greater and less than 3 were 3523 upregulated 4054 downregulated and 2190 upregulated & 2100 downregulated in resistance and susceptible cultivars, respectively. 2-way and and 4-way upregulated and downregulated gene comparisions was carried out. To identify the changes in biological process and molecular function the pathway and gene set enrichment analysis was performed in resistant BR2655 and susceptible HR12 rice cultivars. Conclusions: This study represents the first transcriptome analysis of resistant BR2655 and susceptible HR12 upon M.oryzae infection, generated by RNA-seq technology. This transcriptome analysis further helps to understand the differential gene expression analysis response to the M.oryzae. And candidate genes which are involved in triggering defense mechanism in rice plants. Further it helps to understand the mechanism of activating a cascade of defense responses in resistant and susceptible plants.

Project description:The striped stem borer, Chilo suppressalis (Walker), is a notorious pest of rice that causes large losses in China. Breeding and screening of resistance rice cultivars are effective strategies for C. suppressalis management. In this study, insect-resistant traits of 47 rice cultivars were investigated by C. suppressalis artificial infestation (AI) both in field and greenhouse experiments, using the susceptible (S) cultivar 1665 as a control. Results suggest that two rice cultivars, namely 1688 and 1654, are resistant (R) and moderately resistant (MR) to C. suppressalis, respectively. Then, a comparative transcriptome (RNA-Seq) was de novo assembled and differentially expressed genes (DEGs) with altered expression levels were investigated among cultivars 1688, 1654, and 1665, with or without C. suppressalis infestation for 24 h. A total of 2569 and 1861 genes were up-regulated, and 3852 and 1861 genes were down-regulated in cultivars 1688 and 1654, respectively after artificial infestation with C. suppressalis compared to the non-infested control (CK). For the susceptible cultivar 1665, a total of 882 genes were up-regulated and 3863 genes were down-regulated after artificial infestation with C. suppressalis compared to the CK. Twenty four DEGs belong to proteinase inhibitor, lectin and chitinase gene families; plant hormone signal transduction and plant-pathogen interaction pathways were selected as candidate genes to test their possible role in C. suppressalis resistance. RT-qPCR results revealed that 13 genes were significantly up-regulated and 8 were significantly down-regulated in the resistant cultivar 1688 with C. suppressalis artificial infestation (1688AI) compared to the CK. Three genes, LTPL164, LTPL151, and LOC Os11g32100, showed more than a 10-fold higher expression in 1688AI than in 1688CK, suggesting their potential role in insect resistance. Overall, our results provide an important foundation for further understanding the insect resistance mechanisms of selected resistant varieties that will help us to breed C. suppressalis resistant rice varieties.

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Comparative phosphoproteomic analysis of blast resistant and susceptible rice cultivars in response to salicylic acid.

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Comparative phosphoproteomic analysis of blast resistant and susceptible rice cultivars in response to salicylic acid.

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