Project description:Late blight, caused by the oomycete Phytophthora infestans, is one of the most damaging potato diseases. Genetic resistance is one of the most effective means to control the destruction caused by this pathogen. Transgenic potato lines harboring a resistance gene, RB, confer broad-spectrum, rate-reducing late blight resistance. A microarray approach was used to understand what genes are manipulated in the potato background after the addition of the RB gene that contribute to the late blight resistant phenotype. Keywords: Time course, disease state analysis
Project description:Potato Late blight is one the most important crop diseases worldwide. Even though potato has been studied for many years, the potato disease late blight still has a huge negative effect on the potato production. A total of three commercially available field potato cultivars of different resistance to late blight infection: Kuras (moderate), Sarpo Mira (highly resistant) and Bintje (very suseptable) under controlled green house growing conditions innoculated with a diversity of P. infestans populations. We used label-free quantitative proteomics to investigate the infection with P. infestans in a time-course study over 258 hours. Several key issues limits proteome analysis of potato leaf tissue4–6. Firstly, the immense complexity of the plant proteome which is further complicated by the presence of highly abundant proteins, such as ribulose bisphosphate carboxylase/oxygenase (RuBisCO). Secondly, plant leaf and potato in particular contain abundant levels amounts of phenols and polyphenols which hinder or, unless precautions are taken, completely prevent a successful protein extraction.
Project description:Late blight, caused by the oomycete Phytophthora infestans, is one of the most damaging potato diseases. Genetic resistance is one of the most effective means to control the destruction caused by this pathogen. Transgenic potato lines harboring a resistance gene, RB, confer broad-spectrum, rate-reducing late blight resistance. A microarray approach was used to understand what genes are manipulated in the potato background after the addition of the RB gene that contribute to the late blight resistant phenotype. Keywords: Time course, disease state analysis CRD (3x2x2) Split-Split Plot: 3 sampling time points after inoculation (2, 5, 10 hours), Two genotypes (Katahdin with and without the RB gene), Inoculation with P. infestans or mock inoculation with water. 48 arrays were hybridized in total; 12 in each biological replicate. Each genotype with the mock and late blight inoculated samples was hybridized on two arrays using a dye-swap procedure. Each genotype had a total of 6 arrays across the three sampling time points.
Project description:Potato (Solanum tuberosum) late blight disease caused by Phytophthora. infestans is one of the most critical crop diseases in the world. We investigated the proteome of Solanum tubersoum (cv. Sarpo Mira) by quantitative proteomics after foliar application of encysted zoospore suspension from P. infestans isolate. Also, we examined changes in protein abundance of cv. Sarpo Mira in response to P. infestans infection at three key timepoints 0 hpi (L), 48 hpi (M) and 120 hpi (H), divided into early and late disease stages and analyzed the protein-protein interaction during the disease stages. In response to P. infestans oomycete infection, the iTRAQ comparative proteome survey revealed a total of 1229 differentially expressed proteins (DEPs) in the pairwise comparison of disease stages, which included DEPs specific to early disease stage, DEPs specific to late disease stage and commonly shared DEPs. Over 80% of the change in protein abundance were up-regulated in the early stages of infection while about 61% of the DEPs in the later disease stage were down-regulated. The analysis of expression patterns, functional category, and enrichment tests highlighted significant coordination and enrichment of cell wall-associated defense response proteins in the early stages and cellular protein modification process as well as membrane protein complex formation in the late stages. Furthermore, strong protein interactions were observed in the late disease stage that might relate to disease pathogenesis.
Project description:Identification of novel potato candidate genes for quantitative resistance to Phytophthora infestans by SuperSAGE transcriptome analysis