title: Germ-line transformation and RNAi of the ladybird beetle, Harmonia axyridis.
abstract: To elucidate the molecular mechanisms underlying the tremendous diversity of insect wing colour patterns, it is imperative to identify and functionally characterize the genes involved in this developmental process. Here we report the first successful germ-line transformation using the transposable element vector piggyBac in the ladybird beetle Harmonia axyridis, which demonstrates typical genetic polymorphism in its wing colour patterns. The transformation efficiency by piggyBac was 3.7% per fertile G(0). We investigated the effectiveness of RNAi in Harmonia by injecting EGFP (enhanced green fluorescent protein) dsRNA into early transgenic EGFP-expressing embryos and observed substantial reduction of EGFP fluorescence in 87.2% of hatched larvae. Application of these new genetic tools to non-model insects such as Harmonia will facilitate the broad understanding of developmental mechanisms and evolutionary processes that are inaccessible using established model systems.
authors.affiliation: Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kuwayama
authors.forename: T
authors.initials: T
authors.lastname: Yaginuma
authors.forename: O
authors.initials: O
authors.lastname: Yamashita
authors.forename: T
authors.initials: T
authors.lastname: Niimi
mesh_terms: D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D001517:Coleoptera / Q000235:genetics* / Q000502:physiology; D017931:DNA Primers; D004251:DNA Transposable Elements / Q000235:genetics; D014644:Genetic Variation; D005822:Genetic Vectors; D005854:Germ Cells / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D010858:Pigmentation / Q000235:genetics* / Q000502:physiology; D034622:RNA Interference; D014170:Transformation, Genetic / Q000502:physiology*; D014921:Wings, Animal / Q000502:physiology*
chemical_list: D017931:DNA Primers; D004251:DNA Transposable Elements; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins