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Plant diseases caused by necrotrophic fungi are of world wide economic importance and result in millions of dollars in crop losses annually. In contrast to organisms exhibiting a biotrophic lifestyle, necrotrophic fungi actively kill host tissue prior to colonization usually through the secretion of toxic substances. Necrotrophic diseases are not well controlled by resistant plant cultivars. Modern fungicides can help to control necrotrophic pathogens, but many consider these chemicals to be both environmentally and economically undesirable. The ultimate goal of this project is to identify and develop improved disease resistance strategies for these types of fungal pathogens. This project is focused on the development of an ideal model pathosystem for exploring interactions between plants and necrotrophic fungi. This pathosystem consists of select species found within the plant family Brassicaceae and the necrotrophic, toxin producing fungus, Alternaria brassicicola. This project will further our understanding of host-necrotrophic pathogen relationships in Brassicaceae by first identifying a set of ESTs related to a compatible interaction. A comparative genomics study of pathogen-induced defense responses in resistant and susceptible hosts will be performed using Arabidopsis full-genome microarrays and will give critical insight into the role of specific genes and signaling pathways involved in these responses. A functional genomics approach using knockout mutatgenesis will be performed to elucidate pathogenicity mechanisms in the fungus. A publicly accessible database for this information will be created. This project also includes a major educational component involving an outreach program that will use knowledge and materials obtained from this research as an aid in educating K-12 students about genomics. This project will provide education and scientific training to undergraduates, graduate students, and postdoctoral scientists and will focus on the inclusion of underrepresented groups.
Objective 1: To generate expressed sequence tags (ESTs) from a subtracted cDNA library enriched for genes expressed during a compatible interaction of Brassica oleracea and Alternaria brassicicola. Sequencing of approximately 5,000 ESTs will provide knowledge regarding the susceptible response of a cultivated Brassica species during A. brassicicola infection and a source of in planta-expressed fungal genes.
Objective 2: To dissect fungal pathogenicity mechanisms. Both targeted and random mutagenesis of A. brassicicola will be performed to identify genes important for pathogenicity. The role of a host specific toxin (AB toxin) in pathogenesis and the host factors important for its induction will also be investigated.
Objective 3: To characterize global gene expression patterns in the A. brassicicola-Brassicaceae pathosystem. Microarrays will be used to analyze temporal aspects of global transcriptional profiles of host genes during various compatible and incompatible interactions and in toxin-related studies.
Objective 4: To integrate data and materials from this project into The Science House outreach program. The Science House will act as the primary educational outreach agent in 9 states (including Colorado) by enhancing the current K-12 education component of the NSF-funded Whole genome analysis of pathogen-host recognition and subsequent responses in the rice blast patho-system (NSF No. BDI-0115642) with information, materials and expertise generated from this proposal. This project will also provide education and scientific training for undergraduate and graduate students, technical personnel, and postdoctoral scientists.
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