Abstract
BackgroundStick insects (Phasmatodea) have a high incidence of parthenogenesis and other alternative reproductive strategies, yet the genetic basis of reproduction is poorly understood. Phasmatodea includes nearly 3000 species, yet only the genome of Timema cristinae has been published to date. Clitarchus hookeri is a geographical parthenogenetic stick insect distributed across New Zealand. Sexual reproduction dominates in northern habitats but is replaced by parthenogenesis in the south. Here, we present a de novo genome assembly of a female C. hookeri and use it to detect candidate genes associated with gamete production and development in females and males. We also explore the factors underlying large genome size in stick insects.ResultsThe C. hookeri genome assembly was 4.2 Gb, similar to the flow cytometry estimate, making it the second largest insect genome sequenced and assembled to date. Like the large genome of Locusta migratoria, the genome of C. hookeri is also highly repetitive and the predicted gene models are much longer than those from most other sequenced insect genomes, largely due to longer introns. Miniature inverted repeat transposable elements (MITEs), absent in the much smaller T. cristinae genome, is the most abundant repeat type in the C. hookeri genome assembly. Mapping RNA-Seq reads from female and male gonadal transcriptomes onto the genome assembly resulted in the identification of 39,940 gene loci, 15.8% and 37.6% of which showed female-biased and male-biased expression, respectively. The genes that were over-expressed in females were mostly associated with molecular transportation, developmental process, oocyte growth and reproductive process; whereas, the male-biased genes were enriched in rhythmic process, molecular transducer activity and synapse. Several genes involved in the juvenile hormone synthesis pathway were also identified.ConclusionsThe evolution of large insect genomes such as L. migratoria and C. hookeri genomes is most likely due to the accumulation of repetitive regions and intron elongation. MITEs contributed significantly to the growth of C. hookeri genome size yet are surprisingly absent from the T. cristinae genome. Sex-biased genes identified from gonadal tissues, including genes involved in juvenile hormone synthesis, provide interesting candidates for the further study of flexible reproduction in stick insects.
Highlights
Stick insects (Phasmatodea) have a high incidence of parthenogenesis and other alternative reproductive strategies, yet the genetic basis of reproduction is poorly understood
Genome sequence and analysis The genome size of C. hookeri was estimated at approximately 4.4 Gb using flow cytometry
Sequencing of multiple libraries derived from DNA of a single female (CLI525) yielded 400.4 Gb of PE reads for contig construction with an estimated coverage of 90.9 × the estimated genome size and the initial contigs constructed from these data had a N50 of 3715 bp
Summary
Stick insects (Phasmatodea) have a high incidence of parthenogenesis and other alternative reproductive strategies, yet the genetic basis of reproduction is poorly understood. Clitarchus hookeri is a geographical parthenogenetic stick insect distributed across New Zealand. Clitarchus hookeri is one of the most common New Zealand stick insect species and is distributed across a wide range of habitats on both the North and South Islands with a lower population density at higher latitudes and altitudes [2]. C. hookeri is thought to have hybridized with the obligate parthenogenetic genus Acanthoxyla [4,5,6,7] All these features make C. hookeri an ideal species for the study of geographical parthenogenesis, hybridisation, and mating behaviour [2,3,4,5, 7,8,9,10,11,12,13].
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