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:affy_med_2011_09: In natural ecosystems most vascular plants develop symbiosis with arbuscular mycorrhizal (AM) fungi which help them acquire nutrients such as phosphorus (P) and nitrogen (N). P has long been known to control AM symbiosis which takes place only when P is limiting. For N, however, its role in controlling mycorrhization is less clear. We have chosen the model plant Medicago truncatula to analyze the impact of P limitation and both P and N limitation on Medicago root transcriptome in response to the AM fungus Rhizophagus irregularis (formerly Glomus intraradices (BEG141)). These analyses may help us uncover signaling events involved in the interaction between these symbionts as well as genes encoding transporters potentially important for nutrient exchanges in these conditions. --We will compare the root transcriptome of Medicago truncatula plants inoculated with Rhizophagus irregularis to that of non-inoculated plants grown under P limitation (or both P and N limitation) after 4 weeks of culture
Project description:The nuclei of Medicago truncatula root seedlings exposed to rhizobia were isolated at single-cell resolution with single-nuclei RNA-seq applying 10x Genomics technology.
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_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:Arbuscular mycorrhiza (AM) interactions between plants and Glomeromycota fungi primarily support phosphate aquisition of most terrestrial plant species. To unravel gene expression during early stages of Medicago truncatula root colonization by AM fungi, we used genome-wide transcriptome profiling based on mycorrhizal root fragments enriched for early fungal infection stages. We used Medicago GeneChips to detail the global programme of gene expression in response to early stages of colonization by arbuscular mycorrhizal fungi and identified genes differentially expressed during these early stages. Medicago truncatula GFP-HDEL hairy roots (genotypes A17 and DMI3) were grown in vertically-oriented petri dishes, incubated at 26M-BM-0C and inoculated with 8 Gigaspora margarita spores, which were positioned between the lateral roots. G.margarita spores germinated in 2 to 4 days. Hyphopodia were observed after 5-6 days. Root fragments which reacted to the fungal contact were collected and frozen. Non-inoculated control root fragments were harvested at a comparable age.
Project description:affy_med_2011_09: In natural ecosystems most vascular plants develop symbiosis with arbuscular mycorrhizal (AM) fungi which help them acquire nutrients such as phosphorus (P) and nitrogen (N). P has long been known to control AM symbiosis which takes place only when P is limiting. For N, however, its role in controlling mycorrhization is less clear. We have chosen the model plant Medicago truncatula to analyze the impact of P limitation and both P and N limitation on Medicago root transcriptome in response to the AM fungus Rhizophagus irregularis (formerly Glomus intraradices (BEG141)). These analyses may help us uncover signaling events involved in the interaction between these symbionts as well as genes encoding transporters potentially important for nutrient exchanges in these conditions. --We will compare the root transcriptome of Medicago truncatula plants inoculated with Rhizophagus irregularis to that of non-inoculated plants grown under P limitation (or both P and N limitation) after 4 weeks of culture 12 arrays - Medicago; wt vs mutant comparison
Project description:For transcript analysis of early hypersensitive and susceptible responses of Medicago truncatula to the powdery mildew pathogen, Erysiphe pisi, we compared transcripts from pathogen-inoculated and control (non-inoculated) plants 12 h after infection in resistant (A14), partially resistant (A20), and susceptible (DZA315.16) genotypes. Published in: Medicago truncatula to the powdery mildew 1 and anthracnose pathogens, Erysiphe pisi and Colletotrichum trifolii. Molecular Plant Pathology 8(3):307-319 Keywords: 1 time points and 3 genotypes
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:We used laser-capture microdissection (LCM) to isolate specific cells from the Medicago truncatula nodule meristem (M), the distal infection (DIZ), the proximal infection zone (PIZ), infected cells (IC) and uninfected cells (UIC) from the fixation zone. Based on Medicago GeneChips, we identified the cell- and tissue-specific programm of gene expression in Medicago truncatula root nodules.