Cataclysme subtilisparsata Wehrli , 1932 ( Lepidoptera , Geometridae , Larentiinae ) recognized as bona species – an integrative approach

Based on morphological and molecular analyses of “Cataclysme riguata Hb. var. subtilisparsata Wehrli, 1932” and C. riguata (Hübner, 1813) the former is raised to species rank. C. subtilisparsata is re-described here and compared with C. riguata. Molecular data derived from DNA barcoding are discussed. We illustrate male and female genitalia of both species and present data on life history and morphology of the early stages of C. subtilisparsata.


Introduction
Cataclysme Hübner, [1825] is a small West-Palaearctic genus of moths belonging to the tribe Cataclysmini Herbulot, 1961 (Larentiinae, Geometridae).Viidalepp (2011) and Hausmann and Viidalepp (2012) listed the tribal diagnostic characters.In both checklists of Scoble (1999) and Scoble and Hausmann (2007) seven species are listed for this genus.Recently Viidalepp (2009) and Choi and Stüning (2011) revised the Eastern Palaearctic genus Paraplaneta Warren, 1895, recognizing eight species for Paraplaneta and leaving five species in the genus Cataclysme.All members of Cataclysme are of medium size, their coloration varying from light brownish to dark grey.On the wing upper side all transverse lines are well developed, sinuate or dentate, distinct, and whitish.The coloration of the head, frons and vertex correspond rather well to that of the wings.The palpi are short, reduced in size, the proboscis is well developed and chaetosemata are present as two elongate patches.The abdomen and anal tuft are concolorous with wings.Male antennae are broadly flattened laterally, with very short ciliae, which are bipectinate in the closely related genus Paraplaneta.The forewing R5 is stalked with M1, and the areole is present.In male genitalia the uncus is flat, bifid with projections distally rounded and basally fused (Viidalepp 2009).The male genitalia have a "pseudojuxta" (see streak in Fig. 7a), an autapomorphy of Cataclysme and Paraplaneta within the tribe Cataclysmini (Choi and Stüning 2011).The phallus is long, with termino-lateral spinulose crests ('dorsal-external carina' in Viidalepp 2011) and forked vesica with lines of cornuti (the latter are not present in Paraplaneta).The female genitalia (except for corpus bursae) are strongly sclerotized.The ductus bursae is furrowed, the ostium cleft and fused to sternite A7 (Viidalepp 2011).The species are bivoltine or facultative bivoltine and inhabit Mediterranean macchia such as all kinds of steppe biotopes from forest to open xero-montane steppes.So far as is known the larvae are oligophagous feeding on species of Galium and Asperula (Rubiaceae).
Cataclysme subtilisparsata Wehrli, 1932 was described as a variation of Cataclysme riguata (Hübner 1813) based on two specimens (1 ♂, 1 ♀; collected by Pfeiffer near Maraş in June 1929).Prout (1938) regarded the first as a subspecies of the latter, while Scoble (1999) did not mention that taxon at all.The status of this taxon remained unclear until now, due to the lack of sufficient material.Specimens recently collected in south-east Turkey, however, are allowing an integrative taxonomic revision of this taxon.Breeding experiments were undertaken to obtain more information about larval morphology and bionomic data.Results of morphological (genitalia) and DNA-barcode analyses show C. subtilisparsata to be a distinct species (the sixth of the genus) and not a form or subspecies of C. riguata.

Morphological studies
Specimens were photographed before performing a standard method of dissection (Robinson 1976).Genitalia slides were photographed using Zeiss digital stereomicroscope (ZEISS-SteREO: Discovery.V20).Specimens were identified based on comparison with the syntype and the original description (Wehrli 1932).

DNA amplification and sequencing
PCR amplification and sequencing of 658 bp of COI mtDNA of the three freshly collected specimens of C. subtilisparsata was successful using standard protocols (Ivanova et al. 2006) at the Canadian Centre for DNA Barcoding (CCDB; Guelph), in the framework of the Lepidoptera Campaign of the international Barcode of Life program iBOL (see: www.lepbarcoding.org).

Data analysis
Sequences were aligned using BOLD platform (www.boldsystems.org).For construction of the neighbour-joining tree (using K2P model: Kimura 1980) and for calculation of the genetic distances we used MEGA5 (Tamura et al. 2011).For analysis the DNA barcodes of 17 individuals (of five Cataclysme and one Paraplaneta species) with fragment length >500bp were used.All sequences can be accessed in public projects on the barcode of Life Data Systems (BOLD; data-set DS-Cataclys, www.boldsystems.org;cf.Ratnasingham and Hebert 2007), such as in GenBank (for list of analyzed specimens and their GenBank accession numbers see Appendix).
Male genitalia.Uncus flat, bifid, projections distally rounded.Valva broadly sclerotized at costa, with a rounded lobe and a deep, sub-apical incision.Apical projection thin, with small rounded tip.Juxta narrow and largely reduced, situated between the oval basal parts of the valvae and behind the pseudojuxta (only partly visible in Figs 5a, 7a), saccus well developed, broad.Phallus straight, long and slender with termino-lateral spinulose crests; vesica biforked with numerous cornuti (Figs 5a, b).
Diagnosis.C. subtilisparsata differs from the closely related C. riguata by its slightly larger size.Wingspan in the latter 20-25 mm (n>100) in the former 23-27 mm (n=16), in one specimen from Hakkari, however, only 21 mm.The ground colour is notably lighter, on average.Specimens with darkened basal and medium field never occur in C. riguata.Despite a wide range of variation the transverse lines, especially basal and antemedial lines, are often more zigzagging and thus more reminiscent of C. uniformata (Bellier 1862) than of C. riguata.Furthermore, the forewing cell spots are usually absent in the latter.In male genitalia, pseudojuxta of C. riguata (Fig. 7a) round, in C. subtilisparsata elongate sub-rectangular (Fig. 5a).In female genitalia, ductus bursae of C. riguata (Fig. 8) larger, more robust and more strongly sclerotised than in C. subtilisparsata (Fig. 6) DNA barcoding.Genetic similarity and interspecific distances are shown in the neighbour-joining tree (Fig. 9).Exact distance values are listed in Table 1.Based on these data Calaclysme subtilisparsata is more than 7% divergent from all other examined Calaclysme and Paraplaneta species, confirming our hypothesis of species rank for C. subtilisparsata.However, sequencing of more specimens from northern Iran and all regions of Turkey is highly recommended.Furthermore, the identity of the taxon festivata needs to be investigated (cf.Fig. 9) and its lectotype designated.We consider here the populations from Kyrgyzstan and Uzbekistan as belonging to this taxon.The interpretation of Scoble (1999) (mentioning 'Amur' as locus typicus) is erroneous; Staudinger (1892) clearly states that "the Central Asian populations from Alai, Alexander Mountains, Osch, Usgent, Namangan and Prov.Samarkand" should bear this name.Preliminary data furthermore suggest that there is another taxon forming a separate genetic cluster, so far recorded from Georgia, eastern Turkey and Altai mountains.We do not exclude the possibility that this cluster refers to "Cataclysme riguata elbursica Wagner, 1937".Cataclysme shirniensis Ebert, 1965 (described from N. Afghanistan) is not included in the present study due to the lack of material.Bionomics.Similar to other Cataclysme species, C. subtilisparsata is a bivoltine species.The flight period of the first generation lasts from mid-May to the first third of June.The second brood occurs in July (result of in-vitro breeding experiments by first author and in-vivo by Ralf Fiebig in Nemrut-mountain, pers.comm.).The species inhabits steep, more humid east-and north-facing escarpments and outcrops from 1500-2100 m above sea level.The slopes are mainly covered with stands of thorny cushion plants dominated by xero-montane Acantholimon (Plumbaginaceae) and Astragalus (Fabaceae) mixed with  The species shares its habitat with Ennominae species: Charissa pfeifferi (Wehrli, 1951), Charissa mutilata (Staudinger, 1879) and Gnophos libanotica (Wehrli, 1931).Larva.Full-grown larva (L5) moderately slender, length 3 cm.Ground colour dorsally light green.Head beige.Epistigmatal line fine, whitish.Stigmatal line broad, ivory coloured, with a yellow tinge, indistinct.Stigmata bright yellow, bordered by a fine black margin.The whole body is covered scarcely with fine blackish setae, with small blackish patches at their bases.Ventrum uniform whitish-green (Fig. 11).
Geographic distribution.So far Cataclysme subtilisparsata is known only from the high mountain chains of south-east Turkey from Ceyhan Valley in the west to the mountain ridge south of Van in the east and Mazandaran in north Iran (see Fig 9).

Figure 9 .
Figure 9. Un-rooted neighbour-joining tree based on individuals belonging to six species of the genera Calaclysme and Paraplaneta (calculated using the Kimura 2-parameter model with MEGA 5 (Tamura et al. 2011)).

Table 1 .
(Tamura et al. 2011)ces between six species of the genera Cataclysme and Paraplaneta (in %) (based on COI 5' mt-DNA gene fragments, calculated using the Kimura 2-parameter model with MEGA 6(Tamura et al. 2011)).The distances between C. subtilisparsata and other taxa have shown in bold.