De Novo Transcriptome Sequencing of Serangium japonicum (Coleoptera: Coccinellidae) and Application of Two Assembled Unigenes.

Abstract

The ladybird beetle Serangium japonicum is an important predator of whiteflies. Investigations of the molecular mechanisms of this predatory beetle have been hindered by the scarcity of gene sequence data. To obtain gene sequences for the ladybird beetle and determine differences in gene expression between the summer and winter seasons, paired-end sequencing was performed. Real-time PCR was used to validate differences in Krueppel homolog 1 gene (Kr-h1) mRNA expression in summer vs. winter samples. To determined the diversity of the population, annotated cytochrome c oxidase subunit I gene (COX1) gene fragments were amplified from several ladybird beetle populations. The analysis yielded 191,246 assembled unigenes, 127,016 of which (66.4%) were annotated. These functional annotations of gene sequences are currently available from the National Center for Biotechnology Information (NCBI), and will provide a basis for studying the molecular mechanisms underlying the biological characteristics of S. japonicum. We found a change in expression of ribosome-associated genes across seasons, and postulate that this change is because of seasonal variation in temperature and photoperiod. The differential expression of Kr-h1 suggests that S. japonicum can successfully overwinter because the adults enter diapause. To explain the effects of season on Kr-h1 gene expression, we hypothesize a model in which that a short photoperiod affects the density of Ca2+, the subsequent activity of methyl farnesoate epoxidase and the synthesis of JH, and in turn Kr-h1 gene expression. COX1 annotation was concordant with the morphological ID. The same COX1 sequence was found in the samples from several provinces in China. Therefore, the COX1 sequence is worth further study to distinguish beetle species and populations.