Dataset Information

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.

SUBMITTER: Hu YH

PROVIDER: S-EPMC6945030 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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

Hu Ya Hui YH Liu Yong Y Wei Lin L Chen Hao Tao HT

G3 (Bethesda, Md.) 20200107 1

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 <i ...[more]

PMID: 31722887

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Project description:Background:In recent years, next generation high throughput sequencing technologies have proven to be useful tools for investigations concerning the genomics or transcriptomics also of non-model species. Consequently, ornithologists have adopted these technologies and the respective bioinformatics tools to survey the genomes and transcriptomes of a few avian non-model species. The Common Blackbird is one of the most common bird species living in European cities, which has successfully colonized urban areas and for which no reference genome or transcriptome is publicly available. However, to target questions like genome wide gene expression analysis, a reference genome or transcriptome is needed. Methods:Therefore, in this study two Common Blackbirds were sacrificed, their mRNA was isolated and analyzed by RNA-Seq to de novo assemble a transcriptome and characterize it. Illumina reads (125 bp paired-end) and a Velvet/Oases pipeline led to 162,158 transcripts. For the annotation (using Blast+), an unfiltered protein database was used. SNPs were identified using SAMtools and BCFtools. Furthermore, mRNA from three single tissues (brain, heart and liver) of the same two Common Blackbirds were sequenced by Illumina (75 bp single-end reads). The draft transcriptome and the three single tissues were compared by their BLAST hits with the package VennDiagram in R. Results:Following the annotation against protein databases, we found evidence for 15,580 genes in the transcriptome (all well characterized hits after annotation). On 18% of the assembled transcripts, 144,742 SNPs were identified which are, consequently, 0.09% of all nucleotides in the assembled transcriptome. In the transcriptome and in the single tissues (brain, heart and liver), 10,182 shared genes were found. Discussion:Using a next-generation technology and bioinformatics tools, we made a first step towards the genomic investigation of the Common Blackbird. The de novo assembled transcriptome is usable for downstream analyses such as differential gene expression analysis and SNP identification. This study shows the importance of the approach to sequence single tissues to understand functions of tissues, proteins and the phenotype.

Dataset Information

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

Publications

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

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