<strong>Toward a comprehensive COI DNA barcode library for Swiss Stoneflies (Insecta: Plecoptera) with special emphasis on the genus <em>Leuctra</em></strong>

Abstract

The Swiss Barcode of Life initiative (SwissBOL) aims to inventory the genetic biodiversity in Switzerland using a short DNA sequence. DNA barcoding provides an additional tool for species identification that complements traditional morphological approaches. We report on the establishment of a DNA barcode library for Plecoptera, taxa that are of great importance as bioindicators of water quality and that often present difficulties in species-level identification for larvae and female specimens. Non-destructive DNA extraction, PCR amplification and sequencing of part of the mitochondrial gene Cytochrome Oxidase I (COI) was conducted for 440 individuals (one to eight per species) belonging to 90 species (of the 112 reported from Switzerland). Intra and interspecific distances were calculated and gene trees reconstructed. In most cases, COI was efficient in delimiting stonefly species. Some doubtful specimens were subsequently re-examined and a few misidentifications were found, especially in some problematic groups in the genus Leuctra Stephens, 1836. Larger genetic distances in some species (e.g. Leuctra nigra (Olivier 1811)) indicate the possible presence of sibling species, while in a few cases closely related species are genetically difficult to separate (within the Leuctra fusca species group).

Labiobaetis Novikova &amp; Kluge in Borneo (Ephemeroptera, Baetidae)

“…Compared to the usual distances between different Labiobaetis species in that region and because there is no morphological difference, this distance is surprising, but can be still considered as intraspecific. Ball et al (2005) also reported a case with 6% intraspecific distance in a mayfly in North America and intraspecific K2P distances of more than 3.5% are also not uncommon within Plecoptera (Gill et al 2015, Gattolliat et al 2016.…”

Reinstatement of Leuctra biellensis Festa, 1942 (Plecoptera, Leuctridae)

“…Molecular study: DNA was extracted from specimens stored in the collection of the Museum of Zoology in Lausanne, using a non-destructive method allowing a posteriori morphological identification. 658 bp of the mitochondrial protein-coding gene cytochrome c oxydase subunit I (CO1) was amplified using the primers LCO1490 and HCO2198 (for details see Gattolliat et al 2016).…”

“…All specimens are listed in Table 1. Sequences of Leuctra hippopus Kempny, 1899 and Leuctra pseudorosinae Aubert, 1954 were added to the data matrix as outgroups, as these have the most similar mitochondrial haplotypes according to Gattolliat et al (2016).…”

“…Against the background of global warming and other anthropogenic pressures that are threatening freshwater aquatic biodiversity, Ephemeroptera (mayflies), Plecoptera (stoneflies) and Trichoptera (caddisflies) are considered as particularly vulnerable groups (Tierno de Figueroa et al 2010, Conti et al 2014, Errochdi et al 2014. Between 2013 and 2015, DNA barcode sequences were obtained for 90 of the 112 stonefly species reported from Switzerland (Gattolliat et al 2016). Complementary field searches were done in 2015 and 2016 to elucidate several cases of isolated specimens on the gene tree.…”

“…First results showed high intraspecific genetic distances between some populations preliminary identified as Leuctra nigra (Olivier, 1811). It was hypothesized that two species were involved, one in the North of the Alps and one in the South (Gattolliat et al 2016).…”