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Fruit fly (Diptera: Tephritidae) species identification: a rapid molecular diagnostic technique for quarantine application

Published online by Cambridge University Press:  10 July 2009

K. F. Armstrong ,
C. M. Cameron  and
E. R. Frampton
K. F. Armstrong
Affiliation:
Department of Entomology and Animal Ecology, Lincoln University, PO Box 84, Lincoln, Christchurch, New Zealand:
C. M. Cameron
Affiliation:
Department of Entomology and Animal Ecology, Lincoln University, PO Box 84, Lincoln, Christchurch, New Zealand:
E. R. Frampton
Affiliation:
MAF Regulatory Authority (Plants), Ministry of Agriculture, PO Box 24, Lincoln, Christchurch, New Zealand
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Abstract

New Zealand is currently the only major fruit producing country in the world that is free of economically important fruit flies. As part of the effort to maintain this status, there is a need to supplement quarantine decision-making procedures with a means of rapidly identifying immature life stage infestations to the species level. Here we describe a molecular method that achieves this, using simple restriction patterns of ribosomal DNA (rDNA) as diagnostic markers. The 18S and 18S plus internal transcribed spacer (ITS) regions were amplified from larval DNA by the polymerase chain reaction (PCR). Nineteen species, spanning four genera (including five subgenera of Bactrocera) were analysed. Restriction analysis of the 18S PCR product provided poor resolution, even at the generic level. Digestion of the 18S + ITS PCR product, however, generated thirteen diagnostic haplotypes as defined by the composite restriction patterns from RsaI, Sau3a HaeIII and AluI. No variation was detected at these restriction sites within or between populations. Twenty two restriction enzymes have been screened, but diagnostic RFLPs have yet to been found for six out of the ten Bactrocera (Bactrocera) species; B. passiflorae (Froggatt) cannot be distinguished from B. facialis (Coquillet), nor B. kirki (Froggatt) from B. trilineola (Froggatt) or B. neohumeralis (Hardy) from B. tryoni (Froggatt). Geographic origin could assist in distinguishing the first four species, but the latter pair are very closely related with overlapping origins, hosts and adult morphology. All six species, however, are considered high risk with respect to their likely establishment in New Zealand. Therefore diagnosis based on this molecular technique would support the same quarantine decision. We consider this method could be useful as a diagnostic technique and discuss directions for further development.

Type
Original Articles
Information
Bulletin of Entomological Research , Volume 87 , Issue 2 , April 1997 , pp. 111 - 118
Copyright
Copyright © Cambridge University Press 1997

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