Abstract
Adults of the lady beetle species Harmonia axyridis (Pallas) are bred artificially en masse for classic biological control, which requires egg-laying by the H. axyridis ovary. Development-related genes may impact the growth of the H. axyridis adult ovary but have not been reported. Here, we used integrative time-series RNA-seq analysis of the ovary in H. axyridis adults to detect development-related genes. A total of 28,558 unigenes were functionally annotated using seven types of databases to obtain an annotated unigene database for ovaries in H. axyridis adults. We also analysed differentially expressed genes (DEGs) between samples. Based on a combination of the results of this bioinformatics analysis with literature reports and gene expression level changes in four different stages, we focused on the development of oocyte reproductive stem cell and yolk formation process and identified 26 genes with high similarity to development-related genes. 20 DEGs were randomly chosen for quantitative real-time PCR (qRT-PCR) to validate the accuracy of the RNA-seq results. This study establishes a robust pipeline for the discovery of key genes using high-throughput sequencing and the identification of a class of development-related genes for characterization.
Highlights
RNA-seq provides a far more precise measurement of transcript levels and their isoforms than other methods[12]
We performed quantitative reverse transcription analysis to validate the RNA-seq results. This transcriptome-wide study of mRNAs in the H. axyridis ovary will facilitate the study of the functions of development-related genes and provide potential RNAi targets to accelerate the breeding of H. axyridis for use in biological control
After Illumina Solexa deep sequencing, 21.5 Gb of clean data were obtained from the transcriptome sequencing of the H. axyridis adult ovary
Summary
RNA-seq provides a far more precise measurement of transcript levels and their isoforms than other methods[12]. L transcriptome sequence obtained with RNA-seq, which was considered as paradigm in insects based on the new generation of transcriptome sequencing technology[13]. L, which confirmed that the short-read sequencing platform was feasible in non-model insect transcriptome studies[14]. We applied RNA-seq to obtain ovary transcriptomes from H. axyridis adults in Stages (1–4) (S1–S4), and performed an in depth analysis of these data to detect development-related genes. We performed quantitative reverse transcription (qRT-PCR) analysis to validate the RNA-seq results. This transcriptome-wide study of mRNAs in the H. axyridis ovary will facilitate the study of the functions of development-related genes and provide potential RNAi targets to accelerate the breeding of H. axyridis for use in biological control
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