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Isolation, annotation and applications of expressed sequence tags from the olive fly, Bactrocera oleae

Abstract

The olive fruit fly, Bactrocera oleae, is the major pest of the olive tree. Despite its importance, very little genetic and molecular knowledge is available. The present study is a first attempt to identify and characterize B. oleae expressed sequence tags (ESTs). One hundred and ninety-five randomly selected cDNA clones were isolated and the obtained sequences were annotated through BLASTX similarity searches. A set of 159 unique putative transcripts were functionally assigned using Gene Ontology terms in broad categories of biological process, molecular function and cellular component based on D. melanogaster matches. Moreover, the cytogenetic location of 35 ESTs was determined by in situ hybridization to B. oleae polytene chromosomes. The resulting low-resolution EST map more than doubles the available entry points to the insect’s genome and can assist syntenic comparisons with other distant species. The deduced codon usage of the isolated ESTs suggested a conserved pattern of B. oleae with its closest relatives. Additionally, the comparative analysis of B. oleae ESTs with the homologous D. melanogaster genes led to the development of 17 nuclear EPIC-PCR markers for the amplification of intron sequences of 11 Tephritidae species. Sequencing analysis of several cross-amplified intron sequences revealed a high degree of conservation among Bactrocera species and a varying transferability of the generated markers across the examined genera, suggesting that this method can provide a useful tool for the clarification of phylogenetic relationships among different species, particularly in cases of species complexes.

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Acknowledgments

We thank Dr Komitopoulou at the Biology Department of Athens University (Greece) for the generous gift of the B. oleae cDNA library, which was used for the EST isolation. This research was partially supported by the Pythagoras II grant to KDM from the Greek Ministry of National Education and Religious Affairs and the Graduate Programs of the Biochemistry and Biotechnology Department of the University of Thessaly.

Author information

Correspondence to K. D. Mathiopoulos.

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Communicated by S. Hohmann.

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Tsoumani, K.T., Augustinos, A.A., Kakani, E.G. et al. Isolation, annotation and applications of expressed sequence tags from the olive fly, Bactrocera oleae . Mol Genet Genomics 285, 33–45 (2011). https://doi.org/10.1007/s00438-010-0583-y

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Keywords

  • Tephritidae
  • ESTs
  • Gene Ontology (GO)
  • In situ hybridization
  • EPIC markers
  • Codon usage