Project description:We obtained an Arabidopsis mutant from the Arabidopsis Biological Resource Center stock collection and verified that it was homozygous for a T-DNA insertion in the first exon of ORRM1 (SALK_072648, designated here as orrm1). The homozygous mutant did not show any phenotypic defect when grown under growth room conditions. We examined the organelle transcriptome of the mutant for editing defects because other proteins carrying RIP domains have been shown to be editing factors. We analyzed the plastid RNA editing extent with a new methodology based on RNA-seq. Briefly, total RNA is isolated from leaves and RT-PCR products corresponding to known organelle genes are obtained by using gene-specific primers. The products are mixed in equimolar ratio, sheared, and used as templates to produce an Illumina TruSeq library. This RNA-seq analysis demonstrated that ORRM1 is a plastid editing factor; 12 among 34 plastid sites exhibit a severe reduction of editing extent in the mutant relative to the wild-type
Project description:Using whole genome bisulfite sequencing to provide single-base resolution of DNA methylation status in ros1, idm1, and rdd mutants and identify hypermethylated regions comparing to wild type Arabidopsis plants.
Project description:To investigate the deposition of HTR5 in Arabidopsis, we analysed genome-wide HTR5 density in the wild-type Col-0 by ChIP-seq. We then performed HTR5 occupancy analysis using data obtained from ChIP-seq of 3 different plants including HA-HTR5/Col-0 and Col-0. Col-0 acted as negative control.
Project description:Microbe-associated molecular pattern (MAMP)-triggered immunity (MTI) is the first layer of molecular defense encountered by pathogens when they attempt to infect plants. MTI is dependent on cell surface pattern-recognition receptors (PRR) which act upstream of mitogen-activated protein kinase (MAPK) pathways. Genetic screening and mutant knock-out lines have largely contributed to our knowledge of MTI. However, genetic screening is confined to phenotype-causing mutations and scarcely enables the discovery of redundantly-acting proteins. We sought to discover protein components that contribute in MTI, using a phenotype-independent approach to discern nucleus-localized proteins after MTI induction. We report on the nuclear proteome of Columbia-0 (Col-0) and chitin elicitor receptor kinase 1 (cerk1) mutant plants 15 min after MTI induction. Our approach revealed that MAMP-treated cerk1 plants had many proteins in common with Col-0-treated plants following chitosan treatment. cerk1 plants also manifested several unique proteins that were absent from Col-0 plants when elicited with chitosan indicating that they also perceive chitosan. Detailed analysis of the identified proteins revealed a nuclear accumulation of transcriptional regulators and transcription factors, DNA-modifying enzymes, RNA-binding proteins and ribosomal proteins. No novel MAPKs were found although Receptor for activated C kinase 1, a scaffold protein involved in defense, and a nucleotide binding leucine-rich repeat protein, implicated in resistance to Leptosphaeria maculans, were discovered in the nucleus of chitosan-treated plants and absent from water-treated control plants.
Project description:This experiment was donated by The ELP Project website at elp.ucdavis.edu that was supported in part by the Arabidopsis 2010 project, NSF Division of Molecular and Cellular Biosciences, award 0115109. The study of natural genetic variation for plant disease resistance responses is a complementary approach to utilizing mutants to elucidate genetic pathways. While some key genes involved in pathways controlling disease resistance, and signaling intermediates such as salicylic acid and jasmonic acid, have been identified through mutational analyses, the use of genetic variation in natural populations permits the identification of change-of-function alleles, which likely act in a quantitative manner. Whole genome microarrays, such as Affymetrix GeneChips, allow for molecular characterization of the disease response at a genomics level and characterization of differences in gene expression due to natural variation. Differences in the level of gene expression, or expression level polymorphisms (ELPs), can be mapped in a segregating population to identify regulatory quantitative trait loci (expression QTLs) affecting host resistance responses. In order to identify an appropriate RIL population to map QTL controlling disease resistance responses, we performed a parental survey of 7 different Arabidopsis accessions. We treated vegetatively grown plants with either salicylic acid or a control solution, and harvested the plants at 3 different time points after chemical treatment. We present Affymetrix GeneChip microarray expression data for 3 biological replications of this parental survey. Experimenter name = Dina St. Clair Experimenter phone = (530) 752-1740 Experimenter department = Department of Plant Sciences Experimenter institute = UC Davis Experimenter address = UC Davis Campus Experimenter address = 114 Amundson Hall Experimenter address = Davis, California Experimenter zip/postal_code = CA Experimenter country = USA Keywords: strain_or_line; compound_treatment; time_series