Project description:affy_ralstonia_medicago - Ralstonia solanacearum is the causal agent of the devastating bacterial wilt disease. Its infection process was studied with an in vitro inoculation procedure on intact roots of Medicago truncatula. The pathosystem involved susceptible A17 and resistant F83005.5 M truncatula lines infected with the pathogenic strain GMI1000. The mutant A17 line, Sickle, which showed a resistant phenotype was also part of the experiment. To identify host signaling pathway triggered by R. solanacearum infection with a focus on the involvment of ethylene, we used the Medicago Affymetrix array to monitore the expression profiles and the molecular process associated with initial symptoms development (12hpi) and colonization (72hpi). In order to maximize chances to observe differential gene expression, RNA samples were extracted from the root infection zone (root tips) -Three Medicago truncatula lines, A17, F83005.5 and sickle were inoculated with GMI1000 Ralstonai solanacearum strain (107 cfu/ml). RNA were extracted from root extremities (1 cm above the root tip) at time 0, 12h and 72h post inoculation. Three biological repeats were conducted normal vs disease comparison, time course, 27 arrays - Medicago
Project description:affy_ralstonia_medicago - Ralstonia solanacearum is the causal agent of the devastating bacterial wilt disease. Its infection process was studied with an in vitro inoculation procedure on intact roots of Medicago truncatula. The pathosystem involved susceptible A17 and resistant F83005.5 M truncatula lines infected with the pathogenic strain GMI1000. The mutant A17 line, Sickle, which showed a resistant phenotype was also part of the experiment. To identify host signaling pathway triggered by R. solanacearum infection with a focus on the involvment of ethylene, we used the Medicago Affymetrix array to monitore the expression profiles and the molecular process associated with initial symptoms development (12hpi) and colonization (72hpi). In order to maximize chances to observe differential gene expression, RNA samples were extracted from the root infection zone (root tips) -Three Medicago truncatula lines, A17, F83005.5 and sickle were inoculated with GMI1000 Ralstonai solanacearum strain (107 cfu/ml). RNA were extracted from root extremities (1 cm above the root tip) at time 0, 12h and 72h post inoculation. Three biological repeats were conducted
Project description:Bacterial wilt, caused by the soil-borne bacterium Ralstonia solanacearum, is a lethal disease of mulberry, but the molecular mechanisms of the host resistance responses to R. solanacearum remain unclear. In order to better understand molecular resistance mechanisms to R. solanacearum in mulberry, we set out to define the changes in gene expression of resistance and susceptible mulberry cultivars after inoculation with R. solanacearum. Susceptible cultivar YSD10, resistance cultivar KQ10 and YS283 were inoculation with R. solanacearum, mulberry root samples were collected at 1 dpi and non-treated control in all cultivars. Then we performed RNA-Seq analyses on all mulberry root samples using Illumina HiSeq 2000.
Project description:affy_ralstonia_peeters_medicago - We have identified two essential virulence determinants (GALA7, a type III secretion effector and HpaP, a chaperone-like protein) of Ralstonia solanacearum for the infection and colonisation of the host plant Medicago truncatula. The scope of this project is to identify the GALA7 and HpaP-specific transcriptome alteration. For this purpose wild type and mutant infected root material (13h and 72h postinfection) will be analysed on M. truncatula affymetrix chips. Medicago truncatula (A17 line) are grown in vitro on Farheus medium (with Nitrogen source) plantlets are inoculated with water R. solanacearum wt, gala7 and hpap mutants, and root tips are collected at 13h and 72h postinoculation. Experiment was performed 3 times independently. 4 bacteria conditions x 2 harvest times x 3 biological repeats = 24 samples Keywords: gene knock out,normal vs disease comparison,time course,treated vs untreated comparison 24 arrays - Medicago
Project description:Bacterial wilt caused by Ralstonia solanacearum is a lethal, soil-borne disease of tomato. Control of the disease with chemicals and crop rotation is insufficient, because the pathogen is particularly well adapted for surviving in the soil and rhizosphere. Therefore, cultivar resistance is the most effective means for controlling bacterial wilt, but the molecular mechanisms of resistance responses remain unclear. We used microarrays to obtain the characteristics of the gene expression changes that are induced by R. solanacearum infection in resistant cultivar LS-89 and susceptible cultivar Ponderosa.
Project description:affy_ralstonia_peeters_medicago - We have identified two essential virulence determinants (GALA7, a type III secretion effector and HpaP, a chaperone-like protein) of Ralstonia solanacearum for the infection and colonisation of the host plant Medicago truncatula. The scope of this project is to identify the GALA7 and HpaP-specific transcriptome alteration. For this purpose wild type and mutant infected root material (13h and 72h postinfection) will be analysed on M. truncatula affymetrix chips. Medicago truncatula (A17 line) are grown in vitro on Farheus medium (with Nitrogen source) plantlets are inoculated with water R. solanacearum wt, gala7 and hpap mutants, and root tips are collected at 13h and 72h postinoculation. Experiment was performed 3 times independently. 4 bacteria conditions x 2 harvest times x 3 biological repeats = 24 samples Keywords: gene knock out,normal vs disease comparison,time course,treated vs untreated comparison
Project description:Ralstonia solanacearum is the causal agent of bacterial wilt. Arabidopsis thaliana monogenic resistance to the strain GMI1000 is conferred by the RRS1-R gene, present in the resistant ecotype ND-1 and absent in the susceptible ecotype Col-0. The corresponding protein is recognized by the avirulence protein PopP2 of R. solanacearum. In order to identify the plant proteins involved in the PopP2/RRS1-R perception complex, the screening of a root cDNA library in yeast was performed using PopP2 as bait. One interactor found was called Pip3, and we would like to better understand its biological function. Thereby we choose to compare two RNAi lines with their wild type Nd-1 background, and one KO line with its wild type Col-0 background. Also, two overexpressor lines will be compared with their wilt type Nd-1 background and two with their Col-0 background. 27 samples were used in this experiment.