De novo Transcriptome Analysis of Chinese Citrus Fly, Bactrocera minax (Diptera: Tephritidae), by High-Throughput Illumina Sequencing

PLoS One. 2016 Jun 22;11(6):e0157656. doi: 10.1371/journal.pone.0157656. eCollection 2016.

Abstract

The Chinese citrus fly, Bactrocera minax (Enderlein), is one of the most devastating pests of citrus in the temperate areas of Asia. So far, studies involving molecular biology and physiology of B. minax are still scarce, partly because of the lack of genomic information and inability to rear this insect in laboratory. In this study, de novo assembly of a transcriptome was performed using Illumina sequencing technology. A total of 20,928,907 clean reads were obtained and assembled into 33,324 unigenes, with an average length of 908.44 bp. Unigenes were annotated by alignment against NCBI non-redundant protein (Nr), Swiss-Prot, Clusters of Orthologous Groups (COG), Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes Pathway (KEGG) database. Genes potentially involved in stress tolerance, including 20 heat shock protein (Hsps) genes, 26 glutathione S-transferases (GSTs) genes, and 2 ferritin subunit genes, were identified. These genes may play roles in stress tolerance in B. minax diapause stage. It has previously been found that 20E application on B. minax pupae could avert diapause, but the underlying mechanisms remain unknown. Thus, genes encoding enzymes in 20E biosynthesis pathway, including Neverland, Spook, Phantom, Disembodied, Shadow, Shade, and Cyp18a1, and genes encoding 20E receptor proteins, ecdysone receptor (EcR) and ultraspiracle (USP), were identified. The expression patterns of 20E-related genes among developmental stages and between 20E-treated and untreated pupae demonstrated their roles in diapause program. In addition, 1,909 simple sequence repeats (SSRs) were detected, which will contribute to molecular marker development. The findings in this study greatly improve our genetic understanding of B. minax, and lay the foundation for future studies on this species.

MeSH terms

  • Animals
  • Citrus / parasitology*
  • Databases, Genetic
  • Ecdysone / biosynthesis
  • Female
  • Gene Expression Profiling / methods*
  • Gene Expression Regulation, Developmental
  • Gene Ontology
  • Genes, Insect
  • Heat-Shock Proteins / genetics
  • Heat-Shock Proteins / metabolism
  • High-Throughput Nucleotide Sequencing / methods*
  • Insect Proteins / genetics
  • Insect Proteins / metabolism
  • Male
  • Microsatellite Repeats / genetics
  • Molecular Sequence Annotation
  • Phylogeny
  • Pupa / genetics
  • Sequence Homology, Nucleic Acid
  • Tephritidae / genetics*
  • Transcriptome / genetics

Substances

  • Heat-Shock Proteins
  • Insect Proteins
  • Ecdysone

Grants and funding

This study was financially supported by the National Natural Science Foundation of China (31401742), the China Postdoctoral Science Foundation (2014M552307), and the Fundamental Research Funds for the Central Universities (XDJK2013C149 and 2362015XK04). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.