ABSTRACT: Analysis of transcriptomic profile of pea (Pisum sativum L.) breed Finale nitrogen-fixing nodules (4 week after inoculation) by Massive Analysis of cDNA Ends (MACE).
Project description:Analysis of transcriptomic profile of pea (Pisum sativum L.) breed Frisson nitrogen-fixing nodules (4 week after inoculation) by Massive Analysis of cDNA Ends (MACE).
Project description:Analysis of transcriptomic profile of pea (Pisum sativum L.) breed Sprint-2 nitrogen-fixing nodules (3 week after inoculation) by Massive Analysis of cDNA Ends (MACE).
Project description:Analysis of transcriptomic profile of pea (Pisum sativum L.) breed Sparkle nodulated roots (2 week after inoculation) by Massive Analysis of cDNA Ends (MACE).
Project description:Analysis of transcriptomic profile of pea (Pisum sativum L.) breed NGB1238 nodulated roots (4 week after inoculation) by Massive Analysis of cDNA Ends (MACE).
Project description:The secretion of metabolites by plant roots is a key determinant of microbial growth and colonisation. We have used Pisum sativum and its natural symbiont Rhizobium leguminosarum (it can form N2 fixing nodules on pea roots) to study the natural metabolites secreted by roots. To do this root secretion was harvested from pea plants grown under sterile conditions. This root exudate was then concentrated and used as a sole carbon and nitrogen source for growth of the bacteria in the laboratory. These bacteria were harvested in mid-exponential growth and RNA extracted for microarray analysis. As control cultures the bacteria were grown on 30 mM pyruvate as a carbon source and 10 mM ammonium chloride as a nitrogen source and RNA extracted. Two colour microarrays were performed using root exudate cultures versus pyruvate ammonia grown cultures. This was done in biological triplicate.
Project description:This study aims to compare the potential of standard RNA-sequencing (RNA-Seq) and 3’ massive analysis of c-DNA ends (MACE) RNA-sequencing for the analysis of fresh tissue and describes transcriptome profiling of formalin-fixed paraffin-embedded (FFPE) archival human samples by MACE.
Project description:Rhizobium leguminosarum bv viciae strain 3841 was inoculated onto pea (Pisum sativum) seeds and nodules were harvested at 28 d. The gene expression was compared to free-living bacteria grown on succinate ammonia AMS medium.
Project description:CLE (CLAVATA3/ENDOSPERM SURROUNDING REGION-related) peptides are systemic regulators of legume-rhizobium symbiosis that negatively control the number of nitrogen-fixing nodules. In Medicago truncatula, the expression of the MtCLE35 gene is activated in response to rhizobial inoculation and nitrate treatment, and its overexpression systemically inhibits nodulation. However, little is known about the molecular mechanisms underlying MtCLE35-mediated inhibition of nodulation. In order to elucidate target genes regulated by the MtCLE35-induced signaling cascade, we analyzed the transcriptome of MtCLE35 overexpressing roots inoculated by rhizobia using MACE (Massive Analysis of cDNA Ends) sequencing. Totally, 1390 genes were found to be differentially expressed between MtCLE35-overexpressing (35S::MtCLE35) and control (35S::GUS) roots after rhizobial inoculation, among them 268 genes were upregulated and 1122 genes were downregulated. Among downregulated genes, many known regulators of legume-rhizobia symbiosis were found, where upregulated genes included, among others, genes associated with defense response and cellular redox status. Furthermore, stable transgenic plants overexpressing the MtCLE35 gene were obtained in this study, and gene expression analysis of selected differentially expressed genes in rhizobia inoculated roots was performed in such plants. Collectively, our data suggest that overexpression of the MtCLE35 gene prevents the induction of nodulation program, which is accompanied by the expression of defense-related gene in MtCLE35-overexpressing rhizobia-inoculated M. truncatula roots.
Project description:Plants of the resistant Pisum sativum subsp. syriacum accession P665 and the susceptible pea cultivar Messire were inoculated with M. pinodes.The experiment was conducted in three replicates. 16, 24 and 48 hours after inoculation RNA was isolated from leaves of infected plants and transcribed into cDNA. For each time point and replicate, Cy-labelled cDNA samples from resistant and susceptible plants were mixed and hybridized to Mt16kOLI1Plus microarray