RNA-seq of Medicago truncatula A17 and the ethylene insensitive sickle mutant interacting with the fungal root pathogen, Rhizoctonia solani AG8.
ABSTRACT: This study compares the response of wild type (A17) and ethylene insensitive mutant (sickle) lines of the model legume Medicago truncatula to infection by the root-infecting necrotrophic fungal pathogen, Rhizoctonia solani AG8 (WAC10335). Two time points were taken, 2 and 7 days after inoculation along with corresponding mock-treated controls.
Project description:Paired-end RNA-Seq libraries were constructed for three root sections of the roots of barley cv. Clipper, and landrace Sahara, grown under control and salt-treated (100 mM NaCl) conditions on agar plates in quadruplicate. Experiments were conducted in a temperature-controlled growth cabinet at 17 C in the dark. After three days of germination, seminal roots were dissected according to the following steps: A 1.5 mm long section marked Zone 1 (meristematic zone) was taken from the root tip. A second section (Zone 2) was dissected from the elongation zone up to a third section, Zone 3 (maturation zone), which was excised at the point of visible root hair elongation up to 34 of the entire root. Four biological replicates were generated for each sample in four separate experiments totaling 48 samples. All RNA-seq libraries were constructed and paired-end sequenced (100 bp) on an Illumina HiSeq 2000 system at the Australian Genome Research Facility (Melbourne, Australia).
Project description:Trees establish a symbiotic relationship with specialized soil fungi, called ectomycorrhizae, which is essential for nutrition, growth and health of temperate forest ecosystems. Understanding the mechanisms governing the establishment and functioning of ectomycorrhiza is important because of the role of forests in sequestering CO2 and also to develop ways to optimize tree productivity and sustainability. Here, we investigated the response of an oak species to ectomycorrhiza formation using a two dimensional differential in gel electrophoresis (2D-DIGE) and MALDI-TOF/TOF mass spectrometry proteomics approach. At the root level, changes in the abundance of 34 unique oak proteins were detected and revealed proteins involved in carbon and energy metabolism, protein processing and degradation, response to oxidative stress, lipid metabolism/transport, nitrogen and phosphorous assimilation and cell wall modification. Proteins supporting the importance of the secretory pathway functioning, in particular of the endoplasmic reticulum, during ectomycorrhiza functioning were identified. These proteins were identified as components of the endoplasmic reticulum folding/chaperoning machinery and proteins involved in the ER quality control system. This study constitutes an important contribution for the understanding of the mechanisms underlying the response of plants to ectomycorrhizal symbiosis establishment.
Project description:cortical cell of non-mycorrhizal roots (cor), arbuscule-containing cells (arb) and non-arbuscule-containing cells (nac) of M. truncatula roots colonized with Glomus intraradices were collected by laser capture microdissection (LCM) and used for RNA extraction and Medicago microarray hybridisation
Project description:This experiment was designed to study the interactions between Medicago truncatula and the charcoal rot pathogen Macrophomina phaeolina. Two-week-old plants grown in Magenta boxes supplied with 1/2 MS salt and 1% sucrose were inoculated with M. phaseolina covered wheat seeds, and roots were harvested at 24, 36 and 48 hours after inoculation. Control plants were mock inoculated with a sterile wheat seed, and roots were harvest 24 hours later. Pooled RNAs were used in the array experiment using Affymetrix GeneChip(r) Medicago Genome Array.
Project description:Publication title: Pseudonodule formation by wild type and symbiotic mutant Medicago truncatula in response to auxin transport inhibitors This SuperSeries is composed of the following subset Series: GSE27991: Expression data of Medicago truncatula Jemalong A17 roots treated with auxin transport inhibitors GSE28171: Expression data of Medicago truncatula Jemalong A17 roots treated with S. meliloti exoA mutant or auxin transport inhibitors GSE28172: Expression data of Medicago truncatula skl1-1 roots treated with S. meliloti wild-type or auxin transport inhibitors GSE28173: Genes differentially expressed in wild-type Medicago truncatula plants during nodulation Refer to individual Series
Project description:We used an Affymetrix oligonucleotide microarray consisting of 9,935 tentative consensus (TC)sequences, which are based on cDNA libraries (Mitra et al., 2004 [PMID:15220482]). We examined gene expression of wild-type M. truncatula plants after inoculation with wild-type S. meliloti at 1 d, 4 d, 7d, 14 d, and 21 d. We chose these time points because they span the range of development of the Rhizobium-legume symbiosis, from initiation of the interaction through nitrogen fixation. The gene expression data for the 1-day time point (Mitra et al., 2004 [PMID 15220482]) and the other time points was previously published (Starker el al., 2006 [PMID 16407449]). Three or five independent biological replicates were performed at each time point (1, 4, 7, 14 and 21 days after treatment) and each treatment (buffer and wild-type bacteria).
Project description:Medicago truncatula, which has a relatively small diploid genome, has been adopted as a model species for legume genomics. To enhance its value as a model, we have generated a gene expression atlas that provides a global view of gene expression in all major organ systems of this species, with special emphasis on nodule and seed development.
Project description:To identify host signaling pathways triggered by P. omnivora<br>infection, we used microarrays to monitor the expression profiles<br>and the molecular process associated with initial entry at 3 days post-inoculation and colonization at 5 days post-inoculation