Entomopathogenic Nematode Genome Consortium: Sequencing and Annotation of Heterorhabditis bacteriophora Project Summary: This project aims to sequence the genome of the entomopathogenic nematode Heterorhabditis bacteriophora strain TTO1

Heterorhabditis bacteriophora Project Summary: This project aims to sequence the genome of the entomopathogenic nematode Heterorhabditis bacteriophora strain TTO1 as a representative of 45 other species that are used as biological control agents for insect pests worldwide. This nematode is closely related to the two already sequenced free-living nematodes, Caenorhabditis elegans and C. briggsae, and is a sister taxon to several vertebrate parasitic nematodes, but possesses unique attributes including an obligate mutualistic association with the insect pathogenic bacterium, Photorhabdus luminescens and ability to parasitize model insects including Drosophila melanogaster. Furthermore, the recently sequenced genome of P. luminescens provides unique insights into genes associated with pathogenicity and mutualism. Aside from the significance to biological control, the intellectual merit of the project lies in the comparison between the genomes of H. bacteriophora and C. elegans and C. briggsae which will enhance our understanding of the evolutionary and biological processes in animals involved in mutualism and parasitism. Although, H. bacteriophora is similar in morphology and basic biology to C. elegans, its genome is less than half the size of C. elegans (~40 Mb vs 100 Mb). Thus, the genome sequence of H. bacteriophora will provide a new view of the patterns and processes that shape the genomes in response to mutualism and parasitism, and will identify novel genes and regulatory sequences involved in infection, parasitism, pathogenicity, and mutualism. The project will address questions such as; Are the genes involved in basic biology of H. bacteriophora (e.g., dauer formation and sex determination) the same as those of C. elegans? Did unique genome features arise de novo, via horizontal gene transfer, or by modification of existing genes and pathways? Therefore, the specific objectives of this proposal are to: (i) sequence and annotate the genome of H. bacteriophora and disseminate the information through the WormBase database, (ii) compare H. bacteriophora, C. elegans, C. briggsae, Brugia malayi (a human parasitic nematode), and other in-progress Secernentean genomes for synteny, trans-splicing, regulatory DNA and proteins, and unique genes, and (iii) engage students, scientific community, extension personnel, and general public in the genome project through education, outreach, and workshops. The broader impacts of the project include (i) revolutionization of research in over 100 academic and industrial laboratories developing entomopathogenic nematodes and their symbiotic bacteria as biological control agents worldwide, (ii) facilitation of functional genomic research geared towards the enhancement of infective juvenile longevity, stress tolerance, and virulence that will result in improved efficacy and wider use of nematode in biocontrol, leading to reduced reliance on chemical insecticides, (iii) establishment of H. bacteriophora as a tractable model for the study of parasitism, mutualism, and pathogenicity, (iv) provision of avenues for investigating the obligate tripartite interactions among H. bacteriophora, P. luminescens and insect larvae, including recognition and signaling, leading to immediate and direct applications in agriculture (e.g., targets for pharmacological intervention and genetic manipulation), (v) training of undergraduate, graduate, and post-graduate students including minorities and women in genome annotation, and (vi) enhanced awareness of the value of genome sequencing among extension personnel, farmers, and public. Chances of our success are greatly enhanced as H. bacteriophora shares many attributes that enabled C. elegans become an established model system and techniques for establishing inbred lines, genetic selection, hybridization, mutagenesis, and transgene expression have already been established for both H. bacteriophora and its symbiotic bacterium. We are also well prepared because our team members have experience with genomic projects including C. elegans and C. briggsae.