Deep sequencing analysis of RNAs from a grapevine showing Syrah decline symptoms reveals a multiple virus infection that includes a novel virus

In a search for viruses associated with decline symptoms of Syrah grapevines, we have undertaken an analysis of total plant RNA sequences using Life Sciences 454 high-through put sequencing. 67.5 megabases of sequence data were derived from reverse-transcribed cDNA fragments, and screened for sequences of viral or viroid origin. The data revealed that a vine showing decline symptoms Supported a mixed infection that included seven different RNA genomes. Fragments identified as derived from viruses or viroids spanned a similar to ten thousand fold range in relative prevalence, from 48,278 fragments derived from Rupestris stern pitting-associated virus to 4 fragments from Australian grapevine viroid. 1527 fragments were identified as derived from all unknown marafivirus. Its complete genome was sequenced and characterized, and an RT-PCR test was developed to analyze its held distribution and to demonstrate its presence in leafhoppers (vector for marafiviruses) collected from diseased vines. Initial surveys detected a limited presence of the virus in grape-growing regions of California. (C) 2009 Elsevier Inc. All rights reserved.