TRANSCRIPTOMIC ANALYSIS OF SALIVARY GLANDS AND MIDGUTS FROM BEMISIA TABACI AND THE EFFECTS OF BEGOMOVIRUS INFECTION
Project Number: 5347-22620-021-01
Specific Cooperative Agreement
Start Date: Sep 20, 2010
End Date: Aug 01, 2012
Insect transmission of plant viruses is a major problem in agriculture that requires proactive research to limit its occurrence. The objective of the proposed cooperative research is to better understand the genes expressed in Bemisia tabaci salivary gland and midgut tissues in whiteflies harboring the begomovirus, Squash leaf curl virus. Next-generation massively-parallel DNA sequencing of Bemisia tabaci expressed sequence tag (EST) libraries will provide a more comprehensive representation of genes that are expressed in these tissues than is currently available and how vectoring this begomovirus affects gene expression. These ESTs could represent novel targets for disrupting normal homeostasis of the insect pest and/or viral disease transmission. As many plant disease causing begomoviruses must cross the epithelial barrier within salivary glands or digestive tracts of their arthropod vector, these tissues represent logical targets for understanding the processes involved in virus transmission. How whiteflies respond to viral infection will yield important new information about how begomoviruses enter their insect hosts and are transmitted to plants.
Next-generation sequencing will be used to provide a view of gene expression (e.g. transcriptome) in tissues of Bemisia tabaci vectoring Squash leaf curl virus. The approach follows standard procedures adapted for next-generation sequencing of transcriptomes, including tissue dissection from B. tabaci (naïve and infected with Squash leaf curl virus), RNA extraction, cDNA library construction, massively parallel DNA sequencing of ESTs, annotation, and organization in a PAVE database for mining. Post-sequencing analysis includes verification of changes in gene expression and functional analysis of proteins encoded by transcripts. Documents SCA with U of AZ. Formerly 5347-22620-020-01S (11/10).