Project description:Blumeria graminis f.sp. hordei is an obligate biotrohic fungal pathogen causing powdery mildew in barley. As for other biotrophic fungi, haustorial structures are at the centre of the biotrophic interaction and molecular exchanges, delivering fungal effectors or virulence factors, and taking nutrient from the host. Haustoria are originiated by the fungus, following successful penetration of the initial penetration peg through the plant cell call. Haustorial structures mainly of fungal origin, but they are surrounding by a plant component, the extrauhaustorial membrane and matrix (EHM and EHMx) forming the extrahuastorial complex (EHMc). The plant protein make-up of the plant extrahaustorial components remained unexplored, and this is a first study trying to describe plant proteome associated with haustoria using samples enriched for these structures. Therefore, proteomes of haustoria enriched samples from the epidermis of barley leaves infected with Blumeria graminins f.sp. hordei, the causing agent of barley powdery mildew, were compared to infected epidermis and un-infected epidermis to identify haustoria associated plant proteins. Haustoria were enriched from infected epidermis by digesting epidermal cell walls with cell wall degrading enzymes prior to enrichment for haustorial structures. Proteins identified in these samples were compared to infected and uninfected epidermis samples using a non-targeted label free semi-quantitation method.
Project description:Transcriptional changes were monitored in roots of the barley cultivar Regina following inoculation with zoospores of the adapted plasmodiophorid virus vector Polymyxa graminis using the Affymetrix Barley1 GeneChip®. Barley cv. Regina seeds were imbibed in distilled water over night and then transferred on to moist heat treated (90°C over night) sand in plastic trays that were sealed. Trays were incubated at room temperature for 4 days to allow the seeds to germinate to Zadocks stage 07 of the decimal code for the growth stages of cereals (Tottman et al., 1979 Annals of Applied Biology, 93: 221-234; Zadocks et al., 1974 Weed Research, 14: 415-421). 110 barley (cv. Regina) seedling (Zadocks stage 07) were placed into a 90 mm diameter plastic Petri dish (3 Petri dishes per treatment) and 60 mL of P. graminis zoospore suspension added (1 x 10 6 spores mL-1). Control plants were treated exactly the same way except zoospore-free buffer (0.1 g L-1 Phostrogen, 0.5% Bovine serum albumen) was added. Zoospore challenge (and control) occurred at an ambient temperature of 22°C. Roots were excised from ten plants per treatment at ten time points (15’, 30’, 45’ 1h, 2h, 3h, 4h, 5h, 6h, 7h) and frozen in liquid nitrogen. Three independent biological replicate experiments were done. Total RNA was isolated from each sample using TRI-reagent following the manufacturer’s instructions (Invitrogen) and then treated with DNase I (Ambion, Texas, USA) according to the manufacturer’s protocol. RNA samples for microarray hybridisation were further purified using RNeasy Mini Spin column purification (Qiagen, Hilden, Germany). The integrity of the RNA samples was confirmed using the BioAnalyzer 2100 (Agilent Technologies). Affymetrix GeneChip processing, including RNA quality control, microarray hybridisation and data acquisition was performed through contract research services by Geneservice Ltd (www.geneservice.co.uk). A total of six hybridisations were performed. Publication: European Journal of Plant Pathology 123(1):5-15 ****[PLEXdb(http://www.plexdb.org) has submitted this series at GEO on behalf of the original contributor, Graham RD McGrann. The equivalent experiment is BB74 at PLEXdb.] pathogen isolates: Mock(3-replications); pathogen isolates: Polymyxa graminis(3-replications)
Project description:Waterlogging is a major abiotic stress causing oxygen depletion and carbon dioxide accumulation in the rhizosphere. Barley is more susceptible to waterlogging stress than other cereals. To gain a better understanding of the effect of waterlogging stress in barley, we carried out a genome-wide gene expression analysis in roots of Yerong and Deder2 barley genotypes under waterlogging and control (well-watered) conditions by RNA-Sequencing, using Illumina HiSeq™ 4000 platform.
Project description:We addressed the question how the interaction between the beneficial root endophyte Serendipita vermifera (Sv) and the pathogen Bipolaris sorokiniana (Bs) affects fungal behavior and determines barley host responses using a gnotobiotic natural soil-based split-root system for phenotypic and transcriptional analyses.
Project description:We hypothesized that the genome segments of cultivated barley should show certain similarity with its ancestral wild barley. Instead of whole genome sequences, we employed RNA-Seq to investigated the genomic origin of modern cultivated barley using some representative wild barley genotypes from the Near East and Tibet, and representative world-wide selections of cultivated barley.
Project description:Time course: Interaction of Blumeria graminis f. sp. hordei with Hordeum vulgare, Ingrid (leaf) and Blumeria graminis f. sp. tritici with Hordeum vulgare, Ingrid (leaf)
Project description:Time course: Interaction of Blumeria graminis f. sp. hordei with Hordeum vulgare, Ingrid (leaf epidermis) and Blumeria graminis f. sp. tritici with Hordeum vulgare, Ingrid (leaf epidermis)
