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The redoxproteome: protein nitrosylation and glutathionylation as regulatory mechanisms during plant defense responses to pathogens

ABSTRACT: The present work is directed at studying changes at the proteome level in Arabidopsis thaliana leaves in response to Pseudomonas syringae virulent (Pst) and avirulent (Pst avrRpt2) strains. Arabidopsis leaves were sampled from challenged plants at 4, 8 and 24 hours post inoculation. Proteins were TCA-acetone-phenol extracted and subjected to 2-DE (5-8 pH range) and MS/MS (MALDI-TOF-TOF) analysis. Out of 800 matched spots on each of the 36 gels analysed, 147 spots were either absent in at least one of the conditions studied (time or treatments; qualitative variable spots) or differentially accumulated between time and treatments (quantitative variable spots). Out of the 24 proteins successfully identified over TAIR10 database, 23 have not been reported previously in similar proteomics studies of the Arabidopsis thaliana-Pseudomonas syringae interaction. The exhaustive statistical analysis performed, including principal component and heat map, showed that 24 hours post inoculation can clearly discriminate the challenged plants from the control. The protein change occurred early (4 hours post inoculation) following the virulent pathogen infection, whereas the change occurred later (24 hours post inoculation) following the avirulent pathogen inoculation. Concerning the variable proteins, three behavioural groups can be observed: group 1 (common protein changes in response to virulent and avirulent pathogen infection), group 2 (protein changes in response to virulent pathogen infection) and group 3 (protein changes in response to avirulent pathogen infection). Differential identified proteins following the pathogen infection belonged to different groups including those of oxidative stress defence, enzymes of metabolic pathways and molecular chaperones.

INSTRUMENT(S): instrument model, 4700 Proteomics Analyzer

ORGANISM(S): Arabidopsis Thaliana (mouse-ear Cress)

SUBMITTER: Besma Sghaier-Hammami

PROVIDER: PRD000569 | Pride | 2011-10-19

REPOSITORIES: Pride

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Project description:Transcriptional changes were monitored in the wheat cultivar Avocet*6/Yr1 following inoculation with avirulent and virulent wheat yellow rust (Puccinia striiformis f.sp. tritici) isolates using the Affymetrix wheat genome array GeneChip®. Seedlings of Avocet*6/Yr1 were grown to growth stage 12-13 (Zadoks et al., 1974) before inoculating with either the P.s. f.sp. tritici isolate 169E136 (Yr1-virulent) or 232E137 (Yr1-avirulent) or with talc for the Mock inoculation (Boyd and Minchin 2001). Plants were grown under a 16/8 hour photoperiod cycle, supplemented with sodium lighting (240 ?mol m-2 s-1) at day/night temperatures of 20°C/15°C and a relative humidity of 60%, both before and after inoculation, in a spore-free, containment level 2 greenhouse. Leaf samples from six seedlings were collected 6, 12, 24, 48 and 72 hpi for RNA extraction and transcriptomics analysis from three independent biological experiments. Leaf tissue was ground under liquid nitrogen and total RNA extracted using the TRIzol reagent method, following the manufacturer’s instructions (Invitrogen, Carlsbad, CA). RNA from each time point (16 ?g) were pooled to obtain 80 ?g of total RNA for each treatment (mock inoculated control; avirulent isolate inoculated; virulent isolate inoculated). Pooled RNA samples were further purified using the Qiagen RNeasy mini-kit according to the manufacturer instructions (Qiagen) and RNA integrity was confirmed using the Agilent 2100 Bioanalyzer (Agilent). Affymetrix GeneChip processing, including RNA quality control, microarray hybridisation and data acquisition was performed through contract research services by the John Innes Genome Laboratory, John Innes Centre, Norwich, U.K. A total of nine hybridisations were performed. ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Osman Bozkurt. The equivalent experiment is TA25 at PLEXdb.] pathogen isolates: Mock inoculated (Control)(3-replications); pathogen isolates: Yr1-avirulent wheat yellow rust isolate 232E137(3-replications); pathogen isolates: Yr1-virulent wheat yellow rust isolate 169E136(3-replications)

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The redoxproteome: protein nitrosylation and glutathionylation as regulatory mechanisms during plant defense responses to pathogens

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