Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Expression profiling of a lesion mimic genotype, splNF4050-8, that confers blast resistance in rice

ABSTRACT: Lesion mimic mutants in rice are widely known as spotted leaf (spl) mutants, of which several genotypes exhibit enhanced resistance to different races of Magnaporthe grisea. Besides naturally occurring spotted leaf mutants, tissue culture-induced reverse genetic repositories also act as sources of lesion mimic mutants in rice. We systematically evaluated a large collection of Tos17 mutant panel lines, developed and maintained at the National Institute of Agrobiological Sciences, Tsukuba, Japan, for their reaction to three different races of M. grisea and identified a lesion mimic mutant, NF4050-8, that showed lesions similar to naturally occurring spl5 mutant and enhanced resistance to all three blast races tested. Microarray analysis of ~44,000 rice genes in NF4050-8 with Nipponbare as control during the progressive lesion appearance stage revealed significant up-regulation of numerous defense/pathogenesis-related genes as well as several WRKY domain-containing genes and down-regulation of haem peroxidase gene. Subsequent real-time PCR analysis of WRKY45 and PR1b genes in NF4050-8 and spl5 suggested possible constitutive activation of a defense signaling pathway downstream of Salicylic Acid (SA) but independent of NH1 in these mutant lines of rice. We used Agilent rice oligo microarrays to identify putative defense/pathogenesis-related genes. Leaf tissues of the rice genotypes Nipponbare and NF4050-8 were used in the study. Two replications of microarray experiments were carried out by hybridizing the cRNA from control and progressive lesion appearance stage on 4 x 44k microarray.

ORGANISM(S): Oryza sativa

SUBMITTER: Kameswara Rao Kottapalli

PROVIDER: E-GEOD-17791 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:In this study, integrated transcriptomics, proteomics and metabolomics approaches were applied to investigate the molecular responses of O3 in the leaves of two-weeks old rice (cv. Nipponbare) seedlings exposed to 0.2 ppm O3 for a period of 24 h. Based on the morphological alteration of O3-exposed rice leaves, transcript profiling of rice genes was performed in leaves exposed for 1, 12 and 24 h using rice DNA microarray chip, proteomics and metabolomics. This systematic survey showed that O3 triggers a chain reaction of altered gene, protein and metabolite expressions involved in multiple cellular processes in rice. Also investigated were the molecular responses in the leaves of two-weeks old rice (cv. Nipponbare) seedlings under continuous light and pure air (as a positive control for ozone exposure experiments) for a period of 24 h. Transcript profiling of rice genes was performed in leaves exposed for 1, 12 and 24 h using rice DNA microarray chip, proteomics and metabolomics. This systematic survey showed that continuous light and growth for 24 h also triggers a chain reaction of altered gene expressions involved in multiple cellular processes in rice, but different from those against ozone, in general. Experiment Overall Design: Ozone-treated Samples: An in vivo 14-days-old rice seedling model system was used, where whole seedlings were used for exposure/fumigation to the ozone (0.2 ppm). Dye-swap or reverse labeling with Cy3 and Cy5 dyes procedure was applied followed by hybridization and wash processes, and the hybridized microarrays (G4138A) were scanned using a GenePix microarray scanner followed by the Gene Pix 4000 analysis application program for image analysis and data extraction processes. The GeneSpring Ver. 4 software was used for normalization. Experiment Overall Design: Control Samples: An in vivo 14-days-old rice seedling model system was used, where whole seedlings were used for exposure to continuous light and pure air. Dye-swap or reverse labeling with Cy3 and Cy5 dyes procedure was applied followed by hybridization and wash processes, and the hybridized microarrays (G4138A) were scanned using a Standard protocol of Agilent DNA Microarray Scanner and data processing was done by Feature Extraction software Version 8.1 from Agilent Technologies.

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Expression profiling of a lesion mimic genotype, splNF4050-8, that confers blast resistance in rice

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