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1 December 2006 A Rapid Single-Step Multiplex Method for Discriminating Between Trichogramma (Hymenoptera: Trichogrammatidae) Species in Australia
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Abstract

Inaccurate species identification confounds insect ecological studies. Examining aspects of Trichogramma ecology pertinent to the novel insect resistance management strategy for future transgenic cotton, Gossypium hirsutum L., production in the Ord River Irrigation Area (ORIA) of Western Australia required accurate differentiation between morphologically similar Trichogramma species. Established molecular diagnostic methods for Trichogramma identification use species-specific sequence difference in the internal transcribed spacer (ITS)-2 chromosomal region; yet, difficulties arise discerning polymerase chain reaction (PCR) fragments of similar base pair length by gel electrophoresis. This necessitates the restriction enzyme digestion of PCR-amplified ITS-2 fragments to readily differentiate Trichogramma australicum Girault and Trichogramma pretiosum Riley. To overcome the time and expense associated with a two-step diagnostic procedure, we developed a “one-step” multiplex PCR technique using species-specific primers designed to the ITS-2 region. This approach allowed for a high-throughput analysis of samples as part of ongoing ecological studies examining Trichogramma biological control potential in the ORIA where these two species occur in sympatry.

A. P. Davies, C. L. Lange, and S. L. O’Neill "A Rapid Single-Step Multiplex Method for Discriminating Between Trichogramma (Hymenoptera: Trichogrammatidae) Species in Australia," Journal of Economic Entomology 99(6), (1 December 2006). https://doi.org/10.1603/0022-0493-99.6.2142
Received: 2 May 2006; Accepted: 1 August 2006; Published: 1 December 2006
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