RNA Interference Method for Gene Function Analysis in the Japanese Rhinoceros Beetle Trypoxylus dichotomus

Sakura, Kazuki; Morita, Shinichi; Niimi, Teruyuki

doi:10.21769/bioprotoc.4396

Cited by 1 publication

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“…For the critical period from prepupa to adult, when horn formation occurs, a soil-free breeding system has been established that permits non-invasive continuous monitoring and the precise staging of horn development 12 . Second, an RNA interference (RNAi) technique has been established in T. dichotomus , which allows functional assessment of genes of interest 11 – 17 . Larval RNAi is performed by injecting double-stranded RNA (dsRNA) into the 1st thoracic segment (T1 segment) of the last instar larva just before the prepupal stage.…”

Section: Introductionmentioning

confidence: 99%

The draft genome sequence of the Japanese rhinoceros beetle Trypoxylus dichotomus septentrionalis towards an understanding of horn formation

Morita

Shibata

Nishiyama

et al. 2023

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The Japanese rhinoceros beetle Trypoxylus dichotomus is a giant beetle with distinctive exaggerated horns present on the head and prothoracic regions of the male. T. dichotomus has been used as a research model in various fields such as evolutionary developmental biology, ecology, ethology, biomimetics, and drug discovery. In this study, de novo assembly of 615 Mb, representing 80% of the genome estimated by flow cytometry, was obtained using the 10 × Chromium platform. The scaffold N50 length of the genome assembly was 8.02 Mb, with repetitive elements predicted to comprise 49.5% of the assembly. In total, 23,987 protein-coding genes were predicted in the genome. In addition, de novo assembly of the mitochondrial genome yielded a contig of 20,217 bp. We also analyzed the transcriptome by generating 16 RNA-seq libraries from a variety of tissues of both sexes and developmental stages, which allowed us to identify 13 co-expressed gene modules. We focused on the genes related to horn formation and obtained new insights into the evolution of the gene repertoire and sexual dimorphism as exemplified by the sex-specific splicing pattern of the doublesex gene. This genomic information will be an excellent resource for further functional and evolutionary analyses, including the evolutionary origin and genetic regulation of beetle horns and the molecular mechanisms underlying sexual dimorphism.

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Section: Introductionmentioning

confidence: 99%