Progress 10/01/08 to 09/30/09
Outputs
Progress Report Objectives (from AD-416) Develop a broad spectrum PCR-based method for the detection of a wide range of DMV isolates. Produce antisera to selected DMV proteins and their use in ELISA-based virus detection assay. Evaluate and develop a reliable protocol for meristem tip culture for producing virus-free material. Approach (from AD-416) Based on sequence comparisons of several U.S. isolates and European isolates, PCR primers will be designed that would represent the conserved regions and will be tested for their ability to detect a wide range of DMV isolates. Proteins will be gel-purified and used for immunization. Sera will be collected and tested for their specificity to detect DMV in infected dahlia plants. Using standard tissue culture protocols, meristem culture will be evaluated for its effectiveness in eliminating DMV from cultures that are already infected. Documents Grant with WSU. Formerely 5358-22000-028-05G (8/2007). Significant Activities that Support Special Target Populations Accomplishments for this project include developing a genus-wide PCR- based assay for caulimoviruses including those associated with dahlia, molecular characterization of a new caulimovirus from dahlia (Dahlia common mosaic virus), determination of relative incidence of three caulimoviruses in dahlias in the US, and the identification of caulimoviral sequences in wild species of dahlia from their natural habitats. Information generated is being used to develop virus-specific PCR assays, ELISA-based serological tests, and their use in identification and elimination of virus infected material from propagating stock. Methods of ADODR monitoring included stakeholders meetings, e-mail and phone calls.
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Progress 10/01/06 to 09/30/07
Outputs
Progress Report Objectives (from AD-416) Develop a broad spectrum PCR-based method for the detection of a wide range of DMV isolates. Produce antisera to selected DMV proteins and their use in ELISA-based virus detection assay. Evaluate and develop a reliable protocol for meristem tip culture for producing virus-free material. Approach (from AD-416) Based on sequence comparisons of several US isolates and European isolates, PCR primers will be designed that would represent the conserved regions and will be tested for their ability to detect a wide range of DMV isolates. Proteins will be gel-purified and used for immunization. Sera will be collected and tested for their specificity to detect DMV in infected dahlia plants. Using standard tissue culture protocols, meristem culture will be evaluated for its effectiveness in eliminating DMV from cultures that are already infected. Documents Grant with WSU. Significant Activities that Support Special Target Populations This report serves to document research conducted under a grant agreement between ARS and Washington State University. Additional details of research can be found in the report for the parent project 5358-22000-033- 00D, Integrated Management of Virus Diseases of Small Fruit Crops. Dr. Pappu and collaborators conducted the following research towards the agreements objectives: A survey for the incidence and relative distribution of distinct caulimoviruses associated with dahlia mosaic in the United States and Europe showed that there are at least three distinct caulimoviruses in dahlia and their relative incidence varied considerably. This has important implications for virus testing as material should be tested for all three viruses to avoid propagating and/or distributing infected material. Primers were designed and tested that could detect all three viruses. A refereed journal article describing the results of this survey is in press in Plant Disease. With the information generated on the incidence of different caulimoviruses in dahlia, we completed the complete genome characterization of one of them (DMV-D10). A second distinct caulimovirus is being characterized at the molecular level. We hope to obtain the complete sequence of the genome and conduct comparative studies with other known caulimoviruses. A conserved coding region was identified and protein was expressed in E. coli. Antiserum will be produced to the E. coli-expressed protein. The antiserum will be characterized for its specificity to detect DMV in an ELISA-based assay. Pahalawatta, V., R. Miglino, K.L. Druffel, A. Jodlowska, A. R. van Schadewijk, and H.R. Pappu. 2007. Incidence and relative distribution of distinct caulimoviruses (Genus Caulimovirus, Family Caulimoviridae) associated with dahlia mosaic in Dahlia pinnata. Plant Disease. In press. Pahalawatta, V., K.B. Druffel, S.D. Wyatt, and H.R. Pappu. 2007. Biological and Molecular Studies on Dahlia mosaic virus. Phytopathology 97. In press. Paper to be presented at the annual meeting of the American Phytopathological Society, July 28- August 01, 2007. San Diego, CA. Pappu, H.R. 2007. Virus Diseases of Dahlia. A web page with information on diagnosis and management of viral diseases of dahlia. www.dahlia.wsu.edu. Pappu, H.R. 2007. Dahlia virus research activities. Bulletin of the American Dahlia Society. March 2007. Pp. 78-79. ADODR Statement: The ADODR met with the PI at meetings during the year and discussed results through phone calls and e-mail.
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