2010 Annual Report 1a.Objectives (from AD-416) The long-term objective of this project is to develop a stable, biologically relevant taxonomy for the genus Alternaria with particular reference to the small-spored members of the genus, which are the taxa most often involved in phytosanitary issues and which are also frequently misidentified. Associated with this objective is the development of molecular protocols for detection and identification of target species of phytosanitary importance that are based upon the systematics work. Certain species of Alternaria (e.g., A. gaisen, A. yaliinficiens) are currently known to have a limited geographic distribution that restricts the movement of deciduous tree fruit to the U.S. and other world markets. Other species, many of which are undescribed or frequently misidentified, occur in the United States and in other countries, and need to be characterized and described so that regulatory actions regarding Alternaria on intercepted plant products can be based upon a firm and scientific understanding of the Alternaria taxa that occur on tree fruits and other substrates in the U.S. Over the next 5 years we will focus on the following objective: Objective 1: Isolate and accumulate sequence data from genomic transcripts of known Alternaria spp. which are detected during development of phenotypic traits that vary among different sub-generic groups sensu Simmons and Roberts (1993) and species within those groups, with particular emphasis on sporulation pattern, conidial morphology and, if necessary, germination-mediated metabolite production. 1b.Approach (from AD-416) A fundamental understanding of species-diversity (identity) among Alternaria strains associated with tree fruit will be developed using morphological characters and species-specific DNA sequences identified via subtractive hybridization protocols. Research will be conducted as necessary to address market- or issue-specific questions that arise from interactions with trading partners. Replacing 5350-22000-014-00D (11/03). 3.Progress Report Activities for this project address Problem Statement 1B: Detection, Identification, Characterization, and Classification of Pathogens in the NP 303 Action Plan. Within this problem statement, three sub-problems most accurately describe the current and future research efforts within the project. 1. Although accurately classifying pathogens is important for understanding disease etiology, transmission, and control, the systematics of plant pathogens is plagued by large gaps in knowledge about specific groups of pathogens. 2. Because agricultural globalization has expanded, the need for taxonomic and other biological knowledge of foreign pathogens similarly has increased. 3. Because new molecular approaches may yield accurate tests only when developed within a sound systematic framework, such method development will also require complementary morphological research. ARS scientists characterized small pieces of DNA that are specific for certain species of exotic mold pathogens. This information will allow development of DNA-based tests to determine if exotic pathogens are hitchhiking into the USA on imported commodities. One such piece of DNA is for a gene family known as “aegerolysin”, which we detected in and isolated from the exotic pathogen Alternaria gaisen, which causes a disease called “black spot” of Japanese pear. We have shown this gene sequence is variable enough to be useful for discriminating between some of the most difficult to identify Alternaria species, the ones with small spores.