2012 Annual Report 1a.Objectives (from AD-416): Use a metagenomics approach to identify new viruses infecting the fire ant Solenopsis invicta and subsequently use genomic tools and resources to study the biology (distribution, pathogenicity, etiology, molecular evolutionary history) of several of these viruses. 1b.Approach (from AD-416): 1) Perform a large-scale EST study using 454 pyrosequencing technology to identify potential viruses infecting fire ants using bioinformatics tools and manual annotation of genes. 2) Confirm that newly discovered viruses occur within S. invicta and are not contaminants. 3) Sequence the entire genomes of newly discovered viruses. 4) Use an existing cDNA microarray to determine the host response to viral infections by comparing and identifying genes differentially expressed between virus-infected and uninfected fire ants. 5) Sequence multiple genomes of a single virus and multiple alleles of host genes that respond to viral infection and examine their molecular evolution. 3.Progress Report: These results directly support and correspond to inhouse project objective 1: Develop functional genomic resources and employ these resources to examine the genetic basis of biological traits that can potentially be used for biologically based control, including implications for the geographic origins of infestations; and 2: Expand current biocontrol efforts by discovering and developing new parasites and pathogens; improving mass culture and field release systems; and defining host specificity of natural enemies. The objective of this cooperative research project is to use a metagenomics approach to identify new viruses infecting fire ants and subsequently use genomic tools and resources to study the biology (distribution, pathogenicity, etiology, molecular evolutionary history) of newly discovered viruses. Four cDNA libraries constructed during previous year were sequenced using 454 pyrosequencing technology. Formal analyses of resulting sequence data for viruses using bioinformatics tools revealed two RNA viruses and a single DNA virus, all of which were previously unknown to exist in fire ants. Complete genome sequencing of the DNA virus, along with several additional laboratory and field studies, are underway. Research progress was monitored by by email and telephone correspondence.