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Molecular evidence for a fourth species within the Isotoma viridis group (Insecta, Collembola)
Author(s) -
Burkhardt Ulrich,
Filser Juliane
Publication year - 2005
Publication title -
zoologica scripta
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.204
H-Index - 64
eISSN - 1463-6409
pISSN - 0300-3256
DOI - 10.1111/j.1463-6409.2005.00181.x
Subject(s) - biology , restriction fragment length polymorphism , restriction enzyme , phylogenetic tree , restriction fragment , restriction site , mitochondrial dna , genetics , polymerase chain reaction , cytochrome b , dna , zoology , gene
For identification of single species within the Isotoma viridis group, 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. anglicana and I. 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 of I. viridis into 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 enzymes Mfe I, Nci I and one of Aci I, Bst EII, Nde I, or Sfc I. From the enzymes tested in a previous study, Ase I proved to be reliable, whereas Mbo I can no longer be recommended.