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- Title
Molecular evidence for a fourth species within theIsotoma viridisgroup (Insecta, Collembola).
- Authors
Burkhardt, Ulrich; Filser, Juliane
- Abstract
Burkhardt, U.&Filser, J. (2005). Molecular evidence for a fourth species within theIsotoma viridisgroup (Insecta, Collembola).— Zoologica Scripta, **, ***–***.For identification of single species within theIsotoma viridisgroup, we present polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) as a fast and efficient DNA-based molecular method. We used five PCR primers amplifying the cytochrome oxidase II (COII) region (760 bp) of the mitochondrial DNA. The sequences clearly separated four species (I. viridis,I. riparia,I. anglicanaandI. caerulea) out of samples from Norway, Sweden, Germany and Switzerland. Examination of genetic variation and phylogenetic relationship did not support the separation of two colour pattern forms ofI. viridisinto distinct species. For RFLP, several restriction enzymes were tested for their ability to produce not only species-specific restriction fragment patterns but to discriminate more than one species per enzyme used with as few cleavage sites as possible. Such a design should render a clear fragment pattern when performing a double digest. These demands appear to be fulfilled best by the combination of the restriction enzymesMfeI,NciI and one ofAciI,BstEII,NdeI, orSfcI. From the enzymes tested in a previous study,AseI proved to be reliable, whereasMboI can no longer be recommended.
- Subjects
SPECIES; MOLECULES; COLLEMBOLA; INSECTS; CYTOCHROME oxidase; DNA
- Publication
Zoologica Scripta, 2005, Vol 34, Issue 2, p177
- ISSN
0300-3256
- Publication type
Article
- DOI
10.1111/j.1463-6409.2005.00181.x