Project Number: 5358-21000-037-55
Project Type: Reimbursable

Start Date: Sep 28, 2012
End Date: Sep 27, 2014

Objective:
1. Develop improved virus elimination procedures for the berry crops. 2. Perform virus elimination therapy and produce virus-tested material. 3. Establish a foundation block in a screenhouse with virus-tested planting, retest existing G1 material in the block. 4. Develop laboratory-based diagnostic tests for uncharacterized viruses of the berry crops. 5. Develop new technologies for detection, emphasis will be based on deep sequencing as a method to do broad spectrum testing on clean plants. 6. Develop serological (ELISA) assays for key viruses that are most easily vectored in different regions of the US. 7. Inform nurseries and growers about clean plant activities and promote clean plants and the NCPN activities through a website to reach out to a broader audience, including stakeholders throughout the US.

Approach:
Extracted dsRNAs will be shipped (under permit approval) to the University of Arkansas where they will be converted to cDNA. Sequence analysis will be done at the USDA-ARS-HCRL and University of Arkansas. For deep sequencing both Illumina and 454 technologies will be evaluated for detection of all viruses in a plant. We will also be using Illumina to sequence micro RNAs from plants, since that has been shown recently to be useful in detecting RNA and DNA viruses. Detection primers for RT-PCR will be evaluated at USDA-ARS-HCRL for viruses under permit and at both locations for viruses present in the areas. Virus clean-up will be done at- and virus-free plants will be maintained at USDA-ARS-HCRL.