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Environ Entomol

Title:		Development of a multiplex PCR for identification of vineyard mealybugs
Author(s):		Daane KM; Middleton MC; Sforza R; Cooper ML; Walton VM; Walsh DB; Zaviezo T; Almeida RP;
Address:		"Department of Environmental Science, Policy and Management, University of California, Berkeley, CA 94720-3114, USA. daane@uckac.edu"
Journal Title:		Environ Entomol
Year:		2011
Volume:		40
Issue:		6
Page Number:		1595 - 1603
DOI:		10.1603/EN11075
ISSN/ISBN:		1938-2936 (Electronic) 0046-225X (Linking)
Abstract:		"A simple molecular tool was developed and tested to identify seven mealybug species found in North American vineyards: Pseudococcus maritimus Ehrhorn, Pseudococcus viburni (Signoret), Pseudococcus longispinus (Targioni-Tozzeti), Pseudococcus calceolariae (Maskell), Planococcus ficus (Signoret), Planococcus citri (Risso), and Ferrisia gilli Gullan. The developed multiplex PCR is based on the mitochondrial cytochrome c oxidase subunit one gene. In tests, this single-step multiplex PCR correctly identified 95 of 95 mealybug samples, representing all seven species and collected from diverse geographic regions. To test the sensitivity, single specimen samples with different Pl. ficus developmental stages (egg to adult female and adult male) were processed PCR and the resulting output provided consistent positive identification. To test the utility of this protocol for adult males caught in sex baited pheromone traps, Pl. ficus adult males were placed in pheromone traps, aged at a constant temperature of 26+/-2 degrees C, and processed with the multiplex each day thereafter for 8 d. Results showed consistent positive identification for up to 6 d (range, 6-8 d). Results are discussed with respect to the usefulness of this molecular tool for the identification of mealybugs in pest management programs and biosecurity of invasive mealybugs"
Keywords:		Aging Animals Electron Transport Complex IV/genetics Female Hemiptera/enzymology/*genetics/growth & development Insect Proteins/*genetics Male Molecular Sequence Data Multiplex Polymerase Chain Reaction/*methods North America Nymph/enzymology/genetics/gro;
Notes:		"MedlineDaane, Kent M Middleton, Mathew C Sforza, Rene Cooper, Monica L Walton, Vaughn M Walsh, Douglas B Zaviezo, Tania Almeida, Rodrigo P P eng Evaluation Study Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. England 2012/01/06 Environ Entomol. 2011 Dec; 40(6):1595-603. doi: 10.1603/EN11075"