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Some Orbach Lab Images A genomics approach to understanding pathogenicity of the rice blast fungus, Magnaporthe oryzae Marc Orbach Division of Plant Pathology and Microbiology University of Arizona The Orbach Lab: Using Genetics and Genomics to understand pathogenicity in plant and animal pathogens ? Marc Orbach Division of Plant Pathology and Microbiology Approaches • Mutagenesis, both random and directed to identify pathogenicity genes • Gene expression analysis to identify infection stage-specific genes • Comparative genomics to identify potential conserved pathogenicity factors • GFP reporter genes to observe expression Rice Blast Infection Cycle Attachment Germination 1 hour Sporulation 3-5 days Colonization After 24-48 hours Penetration 24-48 hours Appressorium Development 4-8 hours Melanization http://www.ibwf.de/funagro_index.htm#Model Organism Magnaporthe appressorial mutants generated by insertional mutagenesis Shape defects WT Failure to initiate development Bulbous hyphaewall defects Failure to complete development Surface sensing defect? Mutant testing on rice plants Spray inoculate 14 day rice seedlings (3-4 leaf stage) with 4 mls of conidial suspension (1x105 conidia/ml) Record data at 7 DPI. Cultivar 51583 70-15 : m28 : m32 : m33 : m31 : gelatin WT control Coccidioides - a dimorphic animal pathogen Saprobic phase Parasitic phase • What regulates the dimorphic switch? • What is the environmental niche of the fungus? • What are fungal pathogenicity factors? • Do plant and animal pathogens have common pathogenicity factors? Coccidioides Spherules in vitro Nuclear-localized GFP Coccidioides in vivo spherules Nuclear-localized GFP