Project description:Purpose: The powdery mildew fungus, Blumeria graminis, is an obligate biotrophic pathogen of cereals and has significant impact on food security (Dean et al., 2012). B. graminis f. sp. hordei (Bgh) is the causal agent of powdery mildew on barley (Hordeum vulgare L.). We sought to address the temporal regulation of membrane trafficking associated gene expression in barley-powdery mildew interactions. We created an isogenic panel of immune signaling mutants to address three main questions: (i) which Blumeria secreted proteins are differentially regulated in response to different compromised genotypes, (ii) which barley membrane trafficking genes are altered in response to pathogen attack, and (iii) how are these genes interacting across genotypes and infection stages.
Project description:Purpose: The powdery mildew fungus, Blumeria graminis, is an obligate biotrophic pathogen of cereals and has significant impact on food security (Dean et al., 2012. Molecular Plant Pathology 13 (4): 414-430. DOI: 10.1111/j.1364-3703.2011.00783.x). Blumeria graminis f. sp. hordei (Bgh) is the causal agent of powdery mildew on barley (Hordeum vulgare L.). We sought to identify small RNAs (sRNAs) from both barley and Bgh that regulate gene expression both within species and cross-kingdom.
Project description:Purpose: The powdery mildew fungus, Blumeria graminis, is an obligate biotrophic pathogen of cereals and has significant impact on food security (Dean et al., 2012. Molecular Plant Pathology 13 (4): 414-430. DOI: 10.1111/j.1364-3703.2011.00783.x). Blumeria graminis f. sp. hordei (Bgh) is the causal agent of powdery mildew on barley (Hordeum vulgare L.). We sought to discover novel transcripts expressed following barley infection with blumeria.
Project description:The powdery mildew fungus, Blumeria graminis, is an obligate biotrophic pathogen of cereals and has significant impact on food security (Dean et al., 2012. Molecular Plant Pathology 13 (4): 414-430. DOI: 10.1111/j.1364-3703.2011.00783.x). Blumeria graminis f. sp. hordei (Bgh) is the causal agent of powdery mildew on barley (Hordeum vulgare L.). We sought to identify small RNA-derived transcript cleavage sites from both barley and Bgh that regulate gene expression at the post-transcriptional level both within species and cross-kingdom.
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:We performed RNA-sequencing of Bgh-infected barley leaves at two different time-points after infection to examine gene expression in the barley powdery mildew isolate DH14 during plant pathogenesis.
Project description:Powdery mildew is a very common plant disease and only few plants are immune. Host interactions have been identified and characterized for the pathosystems barley-B. graminis f. sp. tritici (Bgt) and wheat-B. graminis f. sp. hordei (Bgh), whereas no data are reported about powdery mildew and nonhost plants, such as rice. On the other hand rice nonhost resistance is widely unexploited and only few expression data are available. To characterize rice response during nonhost interaction with Bgh, a global expression analysis was performed by using the GeneChip® Rice Genome Array. To describe rice gene expression profiles during nonhost interaction, 2 week-old rice plantlets were inoculated with Bgh. Treated (inoculated) and control (mock) samples were collected 24 hours post-inoculation for GeneChip® Rice Genome Array hybridization.
Project description:Powdery mildew is a very common plant disease and only few plants are immune. Host interactions have been identified and characterized for the pathosystems barley-B. graminis f. sp. tritici (Bgt) and wheat-B. graminis f. sp. hordei (Bgh), whereas no data are reported about powdery mildew and nonhost plants, such as rice. On the other hand rice nonhost resistance is widely unexploited and only few expression data are available. To characterize rice response during nonhost interaction with Bgh, a global expression analysis was performed by using the GeneChipM-BM-. Rice Genome Array. To describe rice gene expression profiles during nonhost interaction, 2 week-old rice plantlets were inoculated with Bgh. Treated (inoculated) and control (mock) samples were collected 24 hours post-inoculation for GeneChipM-BM-. Rice Genome Array hybridization. For transcript proM-oM-,M-^Aling experiments with powdery mildew, entire leaves were sampled from rice plantlets 2 weeks old (cv. Nipponbare). Treated and control (mock) rice leaves were collected 24 hours post inoculation. Three biological replicates for inoculated and control plants were extracted and analysed independently with the GeneChipM-BM-. Rice Genome Array.
Project description:Arabidopsis does not support the growth and asexual reproduction of the barley pathogen, Blumeria graminis f. sp. hordei Bgh). A majority of germlings fail to penetrate the epidermal cell wall and papillae. To gain additional insight into this interaction, we determined whether the salicylic acid (SA) or jasmonate (JA)/ethylene (ET) defence pathways played a role in blocking barley powdery mildew infections. Only the eds1 mutant and NahG transgenics supported a modest increase in penetration success by the barley powdery mildew. We also compared the global gene expression patterns of Arabidopsis inoculated with the non-host barley powdery mildew to those inoculated with a virulent, host powdery mildew, Erysiphe cichoracearum. Genes repressed by inoculations with non-host and host powdery mildews relative to non-inoculated control plants accounted for two-thirds of the differentially expressed genes. A majority of these genes encoded components of photosynthesis and general metabolism. Consistent with this observation, Arabidopsis growth was inhibited following inoculation with Bgh, suggesting a shift in resource allocation from growth to defence. A number of defence-associated genes were induced during both interactions. These genes likely are components of basal defence responses, which do not effectively block host powdery mildew infections. In addition, genes encoding defensins, anti-microbial peptides whose expression is under the control of the JA/ET signalling pathway, were induced exclusively by non-host pathogens. Ectopic activation of JA/ET signalling protected Arabidopsis against two biotrophic host pathogens. Taken together, these data suggest that biotrophic host pathogens must either suppress or fail to elicit the JA/ET signal transduction pathway.
Project description:Attack by the host powdery mildew Erysiphe cichoracearum usually results in successful penetration and rapid proliferation of the fungus on Arabidopsis. By contrast, the nonhost barley powdery mildew Blumeria graminis f. sp. hordei (Bgh) typically fails to penetrate Arabidopsis epidermal cells. In both instances the plant secretes cell wall appositions or papillae beneath the penetration peg of the fungus. Genetic screens for mutations that result in increased penetration of Bgh on Arabidopsis have recently identified the PEN1 syntaxin. Here we examine the role of PEN1 and of its closest homologue, SYP122, identified as a syntaxin whose expression is responsive to infection. pen1 syp122 double mutants are both dwarfed and necrotic, suggesting that the two syntaxins have overlapping functions. Although syp122-1 and the cell wall mur mutants have considerably more pronounced primary cell wall defects than pen1 mutants, these have relatively subtle or no effects on penetration resistance. Upon fungal attack, PEN1 appears to be actively recruited to papillae, and there is a 2-h delay in papillae formation in the pen1-1 mutant. We conclude that SYP122 may have a general function in secretion, including a role in cell wall deposition. By contrast, PEN1 appears to have a basal function in secretion and a specialized defense-related function, being required for the polarized secretion events that give rise to papilla formation.