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Molecular markers for Peristenus spp. (Hymenoptera: Braconidae) parasitoids associated with Lygus spp. (Hemiptera: Miridae)

Published online by Cambridge University Press:  02 April 2012

M. Erlandson
Affiliation:
Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon Saskatchewan, Canada S7N 0X2
L. Braun
Affiliation:
Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon Saskatchewan, Canada S7N 0X2
D. Baldwin
Affiliation:
Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon Saskatchewan, Canada S7N 0X2
J. Soroka
Affiliation:
Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon Saskatchewan, Canada S7N 0X2
M. Ashfaq
Affiliation:
Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon Saskatchewan, Canada S7N 0X2
D. Hegedus*
Affiliation:
Agriculture and Agri-Food Canada, Saskatoon Research Centre, 107 Science Place, Saskatoon Saskatchewan, Canada S7N 0X2
*
1Corresponding author (e-mail: hegedusd@agr.gc.ca).

Abstract

Molecular markers for identifying Peristenus spp. parasitoids to species level and preliminary molecular markers to distinguish two groups of Lygus spp. common to the Canadian prairies were developed. Peristenus species-specific polymerase chain reaction (PCR) primers were developed based on DNA sequence data from a 1600-bp region of the internal transcribed spacer region between the 5.8S and 18S nuclear rRNA genes (ITS2). These primers were able to distinguish Peristenus digoneutis Loan, Peristenus stygicus Loan, and Peristenus pallipes (Curtis). Their ability to identify to species-level parasites dissected from field-collected Lygus spp. nymphs was examined by analysis of DNA from 100 parasite samples. Of those samples showing positive PCR amplification with both control (ITS2) and species-specific primers, all were positive for P. pallipes; none of the samples amplified appropriately sized products with P. digoneutis specific or P. stygicus specific primers. These findings were validated using restriction enzyme digests of amplified regions of the Peristenus spp. cytochrome oxidase 1 gene. Both methods were consistent with earlier studies that showed P. pallipes to be the only species of the genus Peristenus to be associated with Lygus spp. on the Canadian prairies. PCR primers based on DNA sequence data from a 550-bp region of the mitochondrial 16S rRNA gene were designed to discriminate Lyguslineolaris (Palisot de Beauvois) from Lygus borealis (Kelton), and Lygus elisus (Van Duzee). These PCR primers were used to identify field-collected nymphs, with most being identified as either L. borealis/L. elisus (72–82%) orL. lineolaris (14–18%). These estimates of species composition closely reflected those of subsequent adult population surveys from the same fields.

Résumé

Des marqueurs moléculaires capables d'identifier les parasitoïdes Peristenus spp. au niveau spécifique et des marqueurs moléculaires préliminaires capables de distinguer les deux groupes d'espèces de Lygus communes dans les Prairies canadiennes ont été élaborés. Des amorces spécifiques à Peristenus ont été obtenues par la réaction de polymérisation en chaîne (PCR) à partir de séquences d'ADN de 1600 paires de bases dans la région de l'espaceur transcrit interne, entre les gènes 5,8S et 18S de l'ARNr nucléaire (ITS2). Ces amorces sont capables de distinguer Peristenus digoneutis Loan, Peristenus stygicus Loan et Peristenus pallipes (Curtis). Leur capacité d'identifier à l'espèce les parasites trouvés dans des larves de Lygus spp. en nature a été évaluée par analyse de l'ADN de 100 échantillons de parasites. Parmi les échantillons qui ont pu être amplifiés au cours de la procédure PCR, au moyen de l'amorce témoin (ITS2) et des amorces spécifiques, tous correspondaient à P. pallipes et aucun des échantillons n'a amplifié des produits de taille appropriée au moyen des amorces spécifiques à P. digoneutis ou P. stygicus. Ces résultats ont été validés par utilisation d'enzymes de restriction sur les régions amplifiées du gène de la cytochrome oxydase 1 de Peristenus spp. Les deux méthodes donnent des résultats comparables à ceux d'études antérieures qui ont révélé que P. pallipes est la seule espèce de Peristenus à être associée à Lygus spp. dans les Prairies canadiennes. Des amorces PCR basées sur des séquences d'ADN dans une région à 550 paires de bases du gène 16S de l'ARNr mitochondrial ont été conçues pour distinguer Lygus lineolaris (Palisot de Beauvois) de Lygus borealis (Kelton) et de Lygus elisus (Van Duzee). Ces amorces ont servi à identifier des larves récoltées en nature, la plupart appartenant aux espèces L. borealis/L. elisus (72–82%) ou L. lineolaris (12–14%). Ces compositions spécifiques ressemblent de très près à celles qui s'observent dans les populations subséquentes d'adultes dans les mêmes champs.

[Traduit par la Rédaction]

Type
Articles
Copyright
Copyright © Entomological Society of Canada 2003

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