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AMPLIFIED MITOCHONDRIAL DNA AS A DIAGNOSTIC MARKER FOR SPECIES OF CONIFER-FEEDING CHORISTONEURA (LEPIDOPTERA: TORTRICIDAE)

Published online by Cambridge University Press:  31 May 2012

Felix A.H. Sperling  and
Donal A. Hickey
Felix A.H. Sperling
Affiliation:
Department of Biology, University of Ottawa, Ottawa, Ontario, Canada K1N 6N5
Donal A. Hickey
Affiliation:
Department of Biology, University of Ottawa, Ottawa, Ontario, Canada K1N 6N5
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Abstract

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We describe a method for identifying conifer-feeding species and lineages of Choristoneura Lederer in Canada and Alaska. The method relies on amplification of mitochondrial (mt) DNA by the polymerase chain reaction (PCR); amplified DNA is then digested with restriction enzymes to give characteristic DNA fragment patterns. We used the cytochrome oxidase I and II genes of mtDNA, which were previously shown to contain numerous nucleotide differences at the level of species. Ten restriction enzymes were surveyed and a combination of two of these (EcoR V + Hinf I) was sufficient to distinguish the major mtDNA lineages. Choristoneura fumiferana (Clemens), C. pinus Freeman, and C. rosaceana (Harris) were readily distinguished from each other and from an assemblage of three putative western species (C. occidentalis Freeman. C. orae Freeman, and C. biennis Freeman). The three western species have the same mtDNA marker pattern in most individuals and, although ecologically differentiated, their populations may actually be conspecific. At one locality in Alaska, pheromone traps bailed with lures for C. fumiferana attract moths with C. fumiferana mtDNA, and lures for C. orae attract moths with mtDNA that is characteristic of the western assemblage. This demonstrates geographic overlap of genetically distinct species in Alaska. The same two separate mtDNA lineages co-occur at two localities in Alberta, but pheromone attraction is unknown. In British Columbia, populations identified as C. biennis and C. occidentalis contain a few individuals with divergent mtDNA genotypes, the significance of which remains unclear. Amplified mtDNA thus provides a convenient, reliable marker for surveying genetic variation within species and for studying interactions among species of the C. fumiferana group.

Résumé

Nous décrivons ici une méthode d’identification des espèces et des lignées de Choristoneura Lederer parasites des conifères au Canada et en Alaska. La méthode est basée sur l’amplification en chaîne des segments d’ADN mitochondrial (mt) par la polymérase : l’ADN amplifié est ensuite digéré par des enzymes de restriction, ce qui permet de reconnaître les fragments caractéristiques d’ADN. Nous avons utilisé les gènes cytochrome oxydase I et II de l’ADNmt, dans lesquels de nombreux nucléotides diffèrent selon l’espèce. Dix enzymes de restriction ont été utilisés et la combinaison de deux d’entre eux (EcoR V + Hinf I) s’est avérée suffisante pour distinguer la plupart des lignées d’ADNmt. Choristoneura fumiferana (Clemens). C. pinus Freeman et C. rosaceana (Harris) se distinguent facilement l’une de l’autre et se distinguent aussi d’un ensemble de trois espèces probables de l’ouest (C. occidentalis Freeman, C. orae, Freeman et C. biennis Freeman). Les individus des trois espèces de l’ouest possèdent les mêmes fragments d’ADNmt et il est possible que les diverses populations, bien qu’écologiquement différentes, soient conspécifiques. À un endroit en Alaska, des pièges à phéromone ont été garnis de substances propres à attirer des C. fumiferana et ces pièges ont attiré des papillons à ADNmt de C. fumiferana; les pièges garnis de substances propres à attirer des C. orae ont attiré des papillons à ADNmt caractéristique du groupe des espèces de l’ouest. Il y a donc chevauchement géographique d’espèces génétiquement distinctes en Alaska. Les deux mêmes lignées d’ADNmt se retrouvent en deux localités d’Alberta, mais l’effet des phéromones à ces endroits est inconnu. En Colombie-Britannique, les populations identifiées comme C. biennis et C. occidentalis contiennent quelques individus à génotypes d’ADNmt différents, un phénomène qui reste inexpliqué. L’ADNmt amplifié est donc un marqueur commode et fiable dans les études de la variation génétique chez les diverses espèces et des interactions entre espèces au sein du groupe C. fumiferana.

[Traduit par la Rédaction]

Type
Articles
Information
The Canadian Entomologist , Volume 127 , Issue 3 , June 1995 , pp. 277 - 288
Copyright
Copyright © Entomological Society of Canada 1995

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