Review of the genus Kerzhnerocossus Yakovlev, 2011 (Lepidoptera: Cossidae) with descriptions of two new species from Russia and Mongolia Author Saldaitis, Aidas Author Yakovlev, Roman V. Author Truuverk, Andro text Zootaxa 2017 4294 3 389 394 journal article 32538 10.11646/zootaxa.4294.3.9 33e106cc-ce9e-4961-a008-618db6baa57f 1175-5326 840186 78C13075-B932-4424-AFAD-9547F7D013B4 Kerzhnerocossus tannuolus sp. nov. ( Figs 1, 2, 5 , 6, 7 ) Type material . Holotype : male ( Fig. 1 ), S[outhern] Siberia, C [entral] Tuva , Vostochnyi Tannu-Ola Mts. , Durgen r[iver]., Bai-Khaak vill. env., 1250 m , 12̄ 13.vii.2004 , 51°04´N / 94°32´E , leg. J. Hron , M. Česanek & J. Rekelj (slide OP 3497m , GenBank accession code: MF 071456 ) ( Fig. 6 ), (coll.ASV/ WIGJ ). Paratypes : 1 male ( Fig. 2 ), with the same data of locality as the holotype (slide OP 3459m , GenBank accession code: MF 071457 ) ( Fig. 7 ), (coll. OP), 21.vii.2002 , leg. M. Česanek ; 1 male , Russia , Tuva Republic, Tere-Khol lake , sand dunes, 9̄ 12.vi.1995 , 50°01´N 95°03´E , Jalava & Kullberg leg. ( MZH ). Diagnosis. Kerzhnerocossus tannuolus differs from other members of the genus by its larger size (wingspan 28̄ 30 mm vs 24̄ 25 mm ), different forewing pattern and undulated hindwing bands rather than the reticulated pattern as in K. sambainu ( Fig. 4 ) and K. kamelini sp. nov. ( Fig. 3 ). In its male genitalia the valva has a small semicircular process in the center of the costal edge. In K. kamelini ( Fig. 8 ) the process on the costal edge is trapezoidal, the transtilla process is triangular, the angle of the transtilla process apex is about 80° (in K. sambainu ( Fig. 9 ) the angle is about 60°) and the phallus is slightly curved from the base through the middle while in both K. sambainu and K. kamelini the phallus is strongly curved in the middle. FIGURES 1–5. Kerzhnerocossus spp. adults & distribution map. 1. K. tannuolus , male, holotype, [Russia], Tuva, OP3497m (ASV/WIGJ); 2. K. tannuolus , male, paratype, [Russia], Tuva, OP3459m (OP); 3. K. kamelini , male, holotype, Mongolia, Hovd aimak (ZISP); 4. K. sambainu , male, paratype, Mongolia, East aimak (ZISP); 5. Distribution map of the genus Kerzhnerocossus . Description. Wingspan 28̄ 30 mm ( holotype 30 mm ), length of forewing 13̄ 14 mm ( holotype 14 mm ). Head very small, width 0.2 times width of thorax; antenna bipectinate, length of processes exceeding rod diameter by 1.5 times; body densely covered with pale grey scales; forewing cream, with thick reticulated pattern of brown undulated bands and wide transverse bands in the postdiscal and submarginal areas with a series of transverse brown strokes along costal edge, apex rounded; fringe mottled, dark at veins and pale between veins; hindwing cream, with thin reticulated pattern and sputtering of brown scales. Male genitalia ( Fig. 6 ). Uncus triangular, smoothly narrowing to apex, apex blunt, semicircular; gnathos arms thin, long, in fusion form small gnathos with fine spikes on surface; valve relatively narrow with small semicircular process in middle third of costal edge; distal third of valva membranous, transtilla process triangular, angle of transtilla process apex about 80°; juxta small, trapezoidal; saccus small, semicircular; phallus shorter than valve, spoon-like, slightly curved on border of basal and middle thirds; vesica aperture in dorso-apical position half length of the phallus; vesica without cornuti. Female unknown. FIGURES 6–9. Kerzhnerocossus ssp. male genitalia. 6. K. tannuolus , male, holotype, OP3497m (ASV/WIGJ); 7. K. tannuolus , male, paratype, OP3459m (OP); 8. K. kamelini , male, holotype (ZISP); 9. K. sambainu , male, paratype (ZISP). Remarks. One of the paratype specimens has a double uncus ( Fig. 7 ). Despite this unique aberration, the DNA results confirm that the specimen is conspecific with the holotype. There are slight differences in the appearance of the specimens. Paratype from Tere-Khol lake, sand dunes is more light with more expressed postdiscal and submarginal bands. FIGURES 10–11. K. tannuolus biotope. 10. Russia, Tuva, Vostochnyi Tannu-Ola Mts., Durgen river valley general view (photo Martin Česanek, 1997); 11. Russia, Tuva, Vostochnyi Tannu-Ola Mts., Durgen river valley more detailed view (photo Martin Česanek, 1997) Molecular analysis. DNA was extracted from the legs of two K. tannuolus specimens, using a High Pure PCR Template Preparation Kit' (Roche Diagnostics GmbH, Mannheim, Germany). The extraction was carried out following the manufacturer's instructions, with the exception that the first incubation step was 55°C for up to 3 h rather than 1 h. Extracted genomic DNA is housed in the Museum of Natural History of the University of Tartu. Reaction mixtures for PCR and cycle sequencing followed protocol described by Õunap et al . (2016) , with the number of cycles being 45 for PCR, and temperature of the annealing step being 47C for PCR and 45C for cycle sequencing. Sequencing the whole barcode fragment of the COI gene using one primer pair failed. Therefore this gene region was sequenced in two partially overlapping fragments, using primers cov1f (5’-TCG CTT ATT ATT CAG CCA TTT TAT T-3’, Õunap et al. , 2008 ) and cov1r (5’-CTG CAC CAT TTT CTA CAA TTC TTC T-3’, Õunap et al. , 2008 ) for 5’ half, and primers ron (5’-GGA TCA CCT GAT ATA GCA TTC CC-3’, Caterino & Sperling, 1999 ) and nancy (5’-CCC GGT AAA ATT AAA ATA TAA ACT TC-3’, Õunap et al. , 2005 ) for 3’ half. Consensus sequences were created in GENEIOUS R7.1.7 (http://www.geneious.com, Kearse et al . 2012 ) using sequence data from both DNA strands. Sequences were aligned by eye and edited in BIOEDIT 7.2.5 ( Hall 1999 ). Specimen ID with DNA voucher code and GenBank accession code of the COI sequences, are listed as follows: K.tannuolus HT; K.tannuolus 1; MF 071456 and K.tannuolus PT; K.tannuolus 2; MF 071457 . Molecular variation based on the Kimura two-parameter distance model ( Kimura, 1980 ) for 658 bp COI DNA barcodes between two specimens of K. tannuolus was only 0.15 %. Therefore we conclude that the barcoded specimens of K. tannuolus belong to the same species, despite the aberrant genitalia of the paratype . Biology and distribution. Three males were found during the day on 9-12 June 1995 , 12–13 July 2004 and 21 July 2002 in a remote part of Russian Siberia , Tuva region . The species was collected at 1250 meters a.s.l. on the right bank of the Durgen river on north slope of the East Tannu-Ola mountains and sand dunes near Tere-Khol lake . The type locality of K. tannuolus is situated on flat river bank with Larix sibirica Ledeb. forest, with Caragana jubata Pall. shrubs and Salix and Betula (pers. comm. Martin Česanek). Etymology. The new species is named after the type locality.