Morphological and molecular characterisation of the Popijač’s Yellow Sally, Isoperla popijaci sp. nov., a new stenoendemic stonefly species from Croatia (Plecoptera, Perlodidae)

Abstract

A new species of the Yellow Sally genus (Isoperla Banks, 1906) is described, based on morphological (males and females adults, larval and egg) and molecular (the barcode region of the cytochrome c oxidase subunit I gene (COI)) features. Popijač’s Yellow Sally, I. popijaci Hlebec & Sivec, sp. nov. inhabits two karstic sources of the Krasulja rivulet in Croatia. Male and female of the new species are characterised by colouration patterns of the head and pronotum; the dimensions of the female subgenital plate; the medial penial armature and oval-shaped egg without collar and anchor. The larvae differ from their congeners by the uniquely coloured head and pronotum. Based on morphological characteristics I. popijacisp. nov. belongs to the I. tripartita species group. Phylogenetic and taxonomic relationships were reconstructed using three methods of phylogenetic inference and three species delimitation methods. As I. popijacisp. nov. occurs at a narrow area of the Krasulja rivulet in Krbava field, the study puts emphasis on the conservation and hotspot importance of the temporary rivers in the Dinaric karst. Furthermore, the study accentuates the necessity for further research on the genetic diversity of Plecoptera in Croatia.

DNA barcoding for biodiversity assessment: Croatian stoneflies (Insecta: Plecoptera)

“…For samples older than 10 years, the DNA barcode region was amplified as shorter overlapping fragments with primer set: (c) MLepF1/LepR1 and MLepR1/LepF1 ( Hajibabaei et al, 2006 ). All polymerase chain reactions (PCRs) were carried out in a total volume of 20 µl with the PCR mixture prepared following Hlebec et al (2021) . Thermocycling conditions are given in Table S2 .…”