Corresponding authors: Ľuboš Hrivniak (
Academic editor: Eduardo Dominguez
A new species,
Hrivniak Ľ, Sroka P, Bojková J, Godunko RJ, Manko P (2021) A new species of
The global distribution of
Currently, the following species are known from the Caucasus and adjacent areas (
In this study, we provide a detailed morphological investigation and description of the lineage labelled as
This species is distributed in the Hyrcanian forest of southeastern Azerbaijan and northwestern Iran (Fig.
Location of study area (left corner), overall sampling (right corner), and distribution of
Our extensive sampling in the Caucasus and surrounding areas (Fig.
The new species was found to be related to
Although the discovery of distinct lineages represents an essential step for evolutionary studies, their morphological determination is required for practical purposes of nature conservation and biomonitoring surveys. Thus, we aim to complete our phylogenetic studies with morphological investigations and taxonomy of evolutionary lineages delimited by molecular data.
The main aims of this study are to: i) investigate larval morphology of the lineage
The material used for this study was collected by J. Bojková, T. Soldán, and J. Imanpour Namin in Iran (May 2016), and Ľ. Hrivniak, P. Manko, D. Murányi, and M. Žiak in Azerbaijan (September 2018). Larvae were collected by hand net and preserved in 75–96% EtOH. Other
Parts of larval specimens were mounted on microscopic slides using HydroMatrix (MicroTech Lab, Graz, Austria) mounting medium. In order to remove the muscle tissue for an investigation of the cuticular structures, specimens were left overnight in a 10% solution of NaOH prior to slide mounting. Drawings were made using an Olympus SZX7 stereo microscope and an Olympus BX41 microscope, both equipped with a drawing tube. Photographs were obtained using a Leica DFC450 camera on a Leica Z16 APO macroscope and stacked in Helicon Focus v. 5.3 X64. All photographs were subsequently enhanced with Adobe Photoshop CS5. Diagnostic characters for the description of larva were chosen according to
Total genomic DNA of the species was extracted from legs using the DEP-25 DNA Extraction Kit (TopBio s.r.o., Prague, Czech Republic) according to the manufacturer’s protocol. Mitochondrial cytochrome oxidase subunit I (
Species were delimited using the single locus (
Analyses were performed using the SPLITS package for R (
Inter- and intra-specific uncorrected pairwise genetic distances were calculated in MEGA X (Kumar et al. 2018). Distance-based species delimitation was performed using Automatic Barcode Gap Discovery (
Localities of
1 larva (mounted on slide): Azerbaijan, Lənkəran Province, SW of Sim village, unnamed brook (tributary of Digo River);
4 larvae (one barcoded and mounted on slide; labelled N4): Iran: Gilan Province, NW of Sangdeh village, unnamed brook (
1 larva (barcoded; labelled N2): Iran, Gilan Province, S of Tushi village (S of Siahkal village), Shamrud River (RT of Sefid-rud River);
4 larvae (2 barcoded and mounted on slide; labelled CFC4, CFC3): same data as holotype.
1 larva (barcoded; labelled N3): Iran, Gilan Province, W Chelvand village (S of Lavandvil village), Chelavand River (about 2.5 km from its mouth);
The name refers to the distribution of the species in the Hyrcanian forest.
The species is distributed in northwestern Iran and southeastern Azerbaijan (Fig.
General coloration of larvae yellowish-brown, with dark brown to reddish maculation. Body length of male mature larva 8.25 mm (
Unknown.
The main larval diagnostic characters of
The
The intraspecific pairwise genetic distances between specimens of
Although the combination of larval morphological diagnostic characters listed above clearly determine
Coloration pattern of abdominal sterna II–VI makes
Weakly pigmented specimens of
The oblique stripes on abdominal sterna II–VI in
In
Well-defined triangular medial maculae on abdominal terga V–VII are characteristic also for
We are grateful to Ani Bikashvili, Sofia Gabelashvili, Bela Japoshvili, Chris Jonko, Ilhama Kerimova, Dávid Murányi, Jozef Oboňa, Michal Rendoš, Iwona Słowińska, Nataly Snegovaya, and Matej Žiak for their help during the fieldwork. This research was conducted with institutional support (RVO: 60077344) of the Institute of Entomology (Biology Centre, CAS) for LH, PS, and RJG and supported by the Cultural and Educational Grant Agency (The Ministry of Education, Science, Research and Sport of the Slovak Republic) under contract no. 005PU-4/2019 with PM. The sampling campaign in Azerbaijan was supported by the International Visegrad Fund (project no. 21810533).