Location: Horticultural Crops Research
Project Number: 2072-21220-003-01-G
Project Type: Grant
Start Date: Sep 9, 2014
End Date: Sep 30, 2018
Study genetic variability and population genetics of Grapevine leafroll-associated virus 3 for the development of improved diagnostic tools for applications in area-wide clean plant programs and to implement effective management strategies for reducing the spread and economic impact of grapevine leafroll disease in the Pacific Northwest.
Grapevines will be sampled and total nucleic acids will be extracted from petiole samples collected from grapevines showing or suspected for GLD and tested for GLRaV-3 by single tube-one step reverse transcription-polymerase chain reaction (RT-PCR) with primers specific for the heat shock protein 70 homologue (Hsp70h) gene of the virus. The HSP70h and coat protein (CP) genes of representative isolates of GLRaV-3 collected from vineyards will be amplified in RT-PCR assays. The amplified DNA fragments specific to each gene will be cloned and sequenced using standard molecular biology procedures. Based on these results, we will design additional primers to amplify other genomic regions of the virus, besides HSP70h and CP genes, to generate sequence data for determining genetic diversity in GLRaV-3. The derived nucleotide sequence information will be used to determine intra- and inter-vineyard variability as well as cultivar- and/or location-specific variability among the GLRaV-3 using various statistical and bioinformatics techniques. Amendment #3 adds funds to continue the current research objectives and will be used to collect virus isolates and conduct virus detection, cloning, sequencing and sequence analysis of the virus genome.