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Proteomic Analysis of a Rice Mutant sd58 Possessing a Novel d1 Allele of Heterotrimeric G Protein Alpha Subunit (RGA1) in Salt Stress with a Focus on ROS Scavenging.

ABSTRACT: High salinity severely restrains plant growth and results in decrease of crop yield in agricultural production. Thus, it is of great significance to discover the crucial regulators involved in plant salt resistance. Here, we report a novel mutant, sd58, which displays enhanced salt tolerance and dwarf phenotype, by screening from ethyl methane sulfonate (EMS) mutagenized rice mutant library. Genetic analysis showed that sd58 was caused by a single recessive locus. Map-based cloning and allelic test revealed that the phenotypes of sd58 were due to the mutation of RGA1, encoding the alpha subunit of heterotrimeric G protein (G?). A point mutation (G to A) was identified at the splicing site (GT-AG) of the first intron in RGA1, which gives rise to the generation of abnormal mRNA splicing forms. Furthermore, 332 differentially abundant proteins (DAPs) were identified by using an Isobaric Tags for Relative and Absolute Quantitation(iTRAQ)-based proteomic technique from seedlings of sd58 and Kitaake in response to salt treatment. Gene Ontology (GO) and KEGG pathway enrichment analysis revealed these proteins were mainly involved in regulation of the processes such as metabolic pathways, photosynthesis and reactive oxygen species (ROS) homeostasis. Under salt stress, sd58 displayed lower ROS accumulation than Kitaake, which is consistent with the higher enzyme activities involved in ROS scavenging. Taken together, we propose that RGA1 is one of the regulators in salt response partially through ROS scavenging, which might be helpful in elucidating salt tolerant mechanisms of heterotrimeric G protein in rice.

SUBMITTER: Peng P

PROVIDER: S-EPMC6337198 | biostudies-literature | 2019 Jan

REPOSITORIES: biostudies-literature

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Proteomic Analysis of a Rice Mutant sd58 Possessing a Novel d1 Allele of Heterotrimeric G Protein Alpha Subunit (RGA1) in Salt Stress with a Focus on ROS Scavenging.

Peng Peng P Gao Yadi Y Li Zhe Z Yu Yanwen Y Qin Hua H Guo Yan Y Huang Rongfeng R Wang Juan J

International journal of molecular sciences 20190104 1

High salinity severely restrains plant growth and results in decrease of crop yield in agricultural production. Thus, it is of great significance to discover the crucial regulators involved in plant salt resistance. Here, we report a novel mutant, sd58, which displays enhanced salt tolerance and dwarf phenotype, by screening from ethyl methane sulfonate (EMS) mutagenized rice mutant library. Genetic analysis showed that sd58 was caused by a single recessive locus. Map-based cloning ...[more]

PMID: 30621186

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Project description:The genome-wide role of heterotrimeric G-proteins in abiotic stress response in rice has not been examined from a functional genomics perspective, despite the availability of mutants and evidences involving individual genes/processes/stresses. Our rice whole transcriptome microarray analysis (GSE 20925 at NCBI GEO) using the G-alpha subunit (RGA1) null mutant (Daikoku 1 or d1) and its corresponding wild type (Oryza sativa Japonica Nipponbare) identified 2270 unique differentially expressed genes (DEGs). Out of them, we mined for all the potentially abiotic stress-responsive genes using Gene Ontology terms, STIFDB2.0 and Rice DB. The first two approaches produced smaller subsets of the 1886 genes found at Rice DB. The GO approach revealed similar regulation of several families of stress-responsive genes in RGA1 mutant. The Genevestigator analysis of the stress-responsive subset of the RGA1-regulated genes from STIFDB revealed cold and drought-responsive clusters. Meta data analysis at Rice DB revealed large stress-response categories such as cold (878 up/810 down), drought (882 up/837 down), heat (913 up/777 down), and salt stress (889 up/841 down). One thousand four hundred ninety-eight of them are common to all the four abiotic stresses, followed by fewer genes common to smaller groups of stresses. The RGA1-regulated genes that uniquely respond to individual stresses include 111 in heat stress, eight each in cold only and drought only stresses, and two genes in salt stress only. The common DEGs (1498) belong to pathways such as the synthesis of polyamine, glycine-betaine, proline, and trehalose. Some of the common DEGs belong to abiotic stress signaling pathways such as calcium-dependent pathway, ABA independent and dependent pathway, and MAP kinase pathway in the RGA1 mutant. Gene ontology of the common stress responsive DEGs revealed 62 unique molecular functions such as transporters, enzyme regulators, transferases, hydrolases, carbon and protein metabolism, binding to nucleotides, carbohydrates, receptors and lipids, morphogenesis, flower development, and cell homeostasis. We also mined 63 miRNAs that bind to the stress responsive transcripts identified in this study, indicating their post-transcriptional regulation. Overall, these results indicate the potentially extensive role of RGA1 in the regulation of multiple abiotic stresses in rice for further validation.

Dataset Information

Proteomic Analysis of a Rice Mutant sd58 Possessing a Novel d1 Allele of Heterotrimeric G Protein Alpha Subunit (RGA1) in Salt Stress with a Focus on ROS Scavenging.

Publications

Proteomic Analysis of a Rice Mutant <i>sd58</i> Possessing a Novel <i>d1</i> Allele of Heterotrimeric G Protein Alpha Subunit (RGA1) in Salt Stress with a Focus on ROS Scavenging.

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