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Detecting key parasitoids of lepidopteran pests by multiplex PCR

Traugott, Michael; Zangerl, Petra; Juen, Anita; Schallhart, Nikolaus and Pfiffner, Lukas (2006) Detecting key parasitoids of lepidopteran pests by multiplex PCR. Biological Control, 39, pp. 39-46.

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Summary

The use of diagnostic polymerase chain reaction (PCR) is a valuable approach to study host–parasitoid interactions. It circumvents problems inherent to rearing parasitoids from Weld-collected hosts or host dissection. Here, we present a PCR-based detection method for Diadegma semiclausum, Cotesia glomerata, and Cotesia rubecula, which are key parasitoids of the three lepidopteran cabbage pests Plutella xylostella, Pieris brassicae, and Pieris rapae, respectively. Primer pairs for the three parasitoid species and Pl. xylostella were developed: they were used either separately in singleplex PCR or combined in multiplex PCR to (1) screen simultaneously for the two Cotesia species or to (2) detect the parasitoid D. semiclausum and identify its host Pl. xylostella in one PCR. The new idea to simultaneously identify parasitoid and host by molecular markers is useful when the host, in our case early-instar larvae of Pl. xylostella, is morphologically diYcult to distinguish from other host species also occurring in the same habitat. Concentration–response trials revealed comparable detection sensitivity of singleplex and multiplex PCR, with detection limits ranging from 0.03 to 2.2 pg of parasitoid DNA/l PCR. Furthermore, the different developmental time of immature D. semiclausum and C. glomerata did not inXuence parasitoid detection success in either assay type. Based on multiplex PCR screening of Weld-collected caterpillars, we found in Pl. xylostella, P. brassicae, and P. rapae parasitism rates of 33.4% by D. semiclausum, 52% by C. glomerata, and 53.4% by C. rubecula, respectively.
© 2006 Elsevier Inc. All rights reserved.


EPrint Type:Journal paper
Keywords:Plutella xylostella, Diadegma semiclausum, Pieris, Cotesia, Diagnostic PCR, Multiplex PCR, Biological control, Pflanzenschutz, Schädlinge, Nützlinge, Entomologie,
Subjects: Crop husbandry > Crop health, quality, protection
Research affiliation: Switzerland > FiBL - Research Institute of Organic Agriculture Switzerland > Entomolgy
Related Links:http://www.fibl.org/forschung/entomologie/index.php, http://www.elsevier.com/locate/ybcon
Deposited By: Pfiffner, Dr. Lukas
ID Code:13143
Deposited On:07 Mar 2008
Last Modified:12 Apr 2010 07:36
Document Language:English
Status:Published
Refereed:Peer-reviewed and accepted

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