Location: Crop Improvement and Protection Research
Project Number: 2038-22000-013-05
Start Date: Sep 16, 2013
End Date: Mar 15, 2015
• Using established Bemisia tabaci whitefly colonies raised on Brassica oleracea, and a Trialeurodes vaporariorum whitefly colony raised on Physalis wrightii, whiteflies will be transferred virus-infected and healthy tomato plants, respectively, as well as young and old blackberry and Nicotiana benthamiana plants. All experiments will be conducted in triplicate. • RNA will be prepared at Salinas and sent to BTI for preparation RNA-seq and sRNA libraries. • RNA-seq and sRNA sequencing will be multiplexed and conducted on HiSeq at the Cornell Genomic Service Center by BTI. • Bioinformatics analysis on RNA-seq and sRNA (miRNA) will be performed at BTI and genes differentially expressed under comparative conditions will be identified. • Selected RNA expression levels will be evaluated at the USDA-ARS in Salinas from whitefly extract to validate results of RNAseq analysis. • Once whitefly functional genes or their predicted precursor sequences are identified, dsRNAs will be designed and synthesized. • These dsRNAs in varied concentration will be added to artificial diet or allowed uptake into tomato cuttings for ingestion by whiteflies to evaluate impact of each treatment on whitefly mortality and/or impact on reducing virus acquisition and transmission by the whitefly. This work will be conducted at USDA-ARS in Salinas and Charleston. • USDA/ARS-Charleston will provide purified whitefly DNA to BTI for use in genome sequencing. BTI will conduct sequencing, assembly, annotation, and genome evolution and comparative genomics analysis. USDA-ARS-Salinas will contribute funds to partially support a postdoctoral position to be located at BTI for this work.