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Molecular diagnostics of economically important Ceratitis fruit fly species (Diptera: Tephritidae) in Africa using PCR and RFLP analyses

Published online by Cambridge University Press:  09 March 2007

N.B. Barr ,
R.S. Copeland ,
M. De Meyer ,
D. Masiga ,
H.G. Kibogo ,
M.K. Billah ,
E. Osir ,
R.A. Wharton  and
B.A. McPheron
N.B. Barr*
Affiliation:
Department of Entomology, Pennsylvania State University, University Park, PA 16802, USA Center for Plant Health Science and Technology, Pest Detection Diagnostics and Management Laboratory, USDA-APHIS, Moore Air Base, Edinburg, TX 78541, USA
R.S. Copeland
Affiliation:
Department of Entomology, Texas A&M University, College Station, TX 77843, USA International Centre of Insect Physiology and Ecology, PO Box 30772, Nairobi, Kenya
M. De Meyer
Affiliation:
Royal Museum for Central Africa, Entomology Section, Belgium
D. Masiga
Affiliation:
International Centre of Insect Physiology and Ecology, PO Box 30772, Nairobi, Kenya Department of Biochemistry and Biotechnology, Kenyatta University, Nairobi, Kenya
H.G. Kibogo
Affiliation:
International Centre of Insect Physiology and Ecology, PO Box 30772, Nairobi, Kenya
M.K. Billah
Affiliation:
International Centre of Insect Physiology and Ecology, PO Box 30772, Nairobi, Kenya
E. Osir
Affiliation:
International Centre of Insect Physiology and Ecology, PO Box 30772, Nairobi, Kenya
R.A. Wharton
Affiliation:
Department of Entomology, Texas A&M University, College Station, TX 77843, USA
B.A. McPheron
Affiliation:
Department of Entomology, Pennsylvania State University, University Park, PA 16802, USA
*
*Fax: (956) 580 7300 E-mail: [email protected]
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Abstract

The predominantly Afrotropical fruit fly genus Ceratitis contains many species of agricultural importance. Consequently, quarantine of Ceratitis species is a major concern for governmental regulatory agencies. Although diagnostic keys exist for identification of all described Ceratitis species, these tools are based on adult characters. Flies intercepted at ports of entry are usually immatures, and Ceratitis species cannot be diagnosed based on larval morphology. To facilitate identification of Ceratitis pests at ports of entry, this study explores the utility of DNA-based diagnostic tools for a select group of Ceratitis species and related tephritids, some of which infest agriculturally important crops in Africa. The application of the polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method to analyse three mitochondrial genes (12S ribosomal RNA, 16S ribosomal RNA, and NADH-dehydrogenase subunit 6) is sufficient to diagnose 25 species and two species clusters. PCR analysis of the internal transcribed spacer region 1 (ITS-1) is able to distinguish three of the five species left unresolved by mitochondrial DNA analysis.

Keywords

mitochondrial DNAITSCeratitisquarantine
Type
Research Article
Information
Bulletin of Entomological Research , Volume 96 , Issue 5 , October 2006 , pp. 505 - 521
Copyright
Copyright © Cambridge University Press 2006

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