Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression after Verticillium longisporum infection in a susceptible and a resistant A. thaliana near isogenic line

ABSTRACT: Resistance against Verticillium longisporum is quantitative and multigenic. The vec1 QTL in Arabidopsis thaliana controlled resistance against systemic colonization by the fungus. Gene expression was studied to identify genes and pathways controlled by vec1. Near-isogenic lines derived from the resistant ecotype Bur and the susceptible ecotype Ler were compared. NIL9 contained Ler-alleles in the genomic region between 11753 and 12285 kb within vec1 on chromosome 2, tmNIL130 contained Bur-alleles. Microarrays were performed on samples containing the hypocotyl and the shoot basis because colonization is stopped in this region in resistant genotypes. Two developmental stages were studied: time-point (1) at the onset of flowering, when systemic colonization was shown to start and time point (2) at the onset of silique maturity, when extensive fungal proliferation was shown to occur. Two pathogen treatments were studied: (1) mock-inoculated controls, (2) plants inoculated with the V. longipsorum isolate 43 (VL). Gene expression was studied in two genotypes (NIL9-susceptible; tmNIL130-resistant), under two pathogen treatments (mock-inoculated; VL-inoculated) at two time points (at the onset of flowering, 13 days post inoculation [dpi]; at the onset of silique maturity, 27-29 dpi). Three biological replicates were used for each treatment (24 microarrays altogether).

ORGANISM(S): Arabidopsis thaliana

SUBMITTER: Eva Häffner

PROVIDER: E-GEOD-70021 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Detailed information: Rice (*Oryza sativa* L. cv. Nipponbare) is a drought-susceptible species which is well suited for studies of abiotic stress response because of the comprehensive bioinformatics resource available. By withholding water from the entire root system of young rice plants, or half the root system only, it was possible to infer the relative impact of signals arriving from roots growing in wet and dry soil on the shoot proteome. The global proteome of shoots had 685 proteins in common to all three drought treatments but there were major shifts in abundance of individual proteins within 16 functional categories. The dominant changes were analyzed more deeply. First, we investigated transport and cell component organization, where some proteins were up-regulated by drought but many more down-regulated. Proteins involved in protein metabolism were up-regulated in general by drought when they were responsible for protein degradation but those involved in protein synthesis were down-regulated when water was withheld. Stress-related proteins behaved very consistently by increasing in droughted plants but notably some proteins were most abundant when roots of the same plant were growing in both wet and dry soil. This suggests that drought signals are complex interactions and not simply the additive effect of water supply to the roots. Changes in carbohydrate-processing proteins were consistent with the passive accumulation of soluble sugars in shoots under drought, with hydrolysis of sucrose and starch synthesis both enhanced. Data analysis information: The result raw files were converted to mzXML format and processed through the global proteome machine (GPM) software (version 2.1.1) of the X!Tandem algorithm (freely available at http://www.thegpm.org). The 16 gel fractions were processed serially for each experiment and the output files were generated as non-redundant, merged files with protein identifications with log (e) values less than -1, for each individual gel fraction. A protein database compiled from NCBI *O*. *sativa* with 26938 protein sequences (August 2011) was used in GPM to search the tandem mass spectra; the database also included common trypsin and human peptide contaminants. False discovery rates (FDR) were evaluated by searching against a reversed sequence database. Search parameters included MS and MS/MS tolerances of +2 Da and +0.2 Da, carbamidomethylation of cysteine as fixed modifications, oxidation of methionine as variable modifications and tolerance of two missed tryptic cleavages and K/R-P cleavages.

Project description:Bolting is a key process in the growth and development of lettuce (Lactuca sativa L.). High temperature can induce earlier bolting which decreases in both quality and production of lettuce. However, knowledge underlying lettuce bolting is still lacking. To better understand the molecular basis of bolting, a comparative proteomics analysis was conducted on lettuce stems in the bolting period induced by high temperature (33 °C) compared with a control (20 °C) using iTRAQ-based proteomics, phenotypic measures, and biological verifications. High temperature induced lettuce bolting, while control temperature did not. Of the 6656 proteins identified, 758 proteins significantly altered their expression level induced by high-temperature relative to the control, of which 409 were up-regulated and 349 down-regulated. Proteins with abundance level change were mainly involved in photosynthesis, carbohydrate metabolism, stress response, hormone synthesis, and signal transduction. These differential proteins were mainly enriched in pathways associated with photosynthesis and tryptophan metabolism involving in auxin (IAA) biosynthesis. Among the differentially expressed proteins associated with photosynthesis and tryptophan metabolism were up-regulated. Moreover, in gibberellin (GA) biosynthesis pathway, 10 of main enzymes of P450 were up-regulated. Proteins related to SAUR and GRP, implicated in IAA and GA signal transduction were up-regulated, and the phosphorylation and ubiquitination related proteins regulating IAA and GA signal transduction were also induced. These findings indicate that a high temperature enhances the function of photosynthesis, IAA and GA synthesis and signal transduction to promote the process of bolting, which is in line with the physiology and transcription levels of IAA and GA metabolism. Our data provide a first comprehensive dataset for gaining novel understanding of the molecular basis underlying lettuce bolting induced by high temperature. It is potentially important for further functional analysis and genetic manipulation for molecular breeding to breed new cultivar of lettuce to restrain early bolting, which is vital for improving vegetable quality.

Dataset Information

Gene expression after Verticillium longisporum infection in a susceptible and a resistant A. thaliana near isogenic line

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