Oenochroma barcodificata Hausmann & Young

Hausmann, Axel, Hebert, Paul D. N., Mitchell, Andrew, Rougerie, Rodolphe, Sommerer, Manfred, Edwards, Ted & Young, Catherine J., 2009, Revision of the Australian Oenochroma vinaria Guenée, 1858 species-complex (Lepidoptera: Geometridae, Oenochrominae): DNA barcoding reveals cryptic diversity and assesses status of type specimen without dissection, Zootaxa 2239, pp. 1-21 : 7-18

Oenochroma barcodificata Hausmann & Young , sp. nov.

Type material. Holotype: Ƥ, ‘TAS[ MANIA], Chimney Pot Hill, nr Ridgeway, 20 OCT. 1996, eucalypt forest, leg. P.B. McQuillan’, coll. TMAG, genit.slide ‘ ZSM G 13863’, specimen ID ‘ BC ZSM Lep 0 2948 ’ [sequence ID GWORB 1068-07]. Paratypes: coll. BIO: 63 Australia, Tasmania, Hobart, Kingston Beach, lat. -42.986 x long. 147.317, alt. 110 m., 0 2., 11. and 18. XII.2005, 10. X.2006, 27. XII.2006, 22. I.2007, leg. R. D. Ward, sequence IDs: LOTSA 134-06/06-TASA-00134, LOTSA 125-06/06-TASA-00125, LOTSA 122-06/06-TASA- 0 0 122, LOTSC 391-07/06-TASB-01390, LOTSC 814-07/06-TASB-01813, LOTSD 507-08/07-TASB-0507; coll. M. Sommerer: 1Ƥ, Tasmania sept., vic. Mt. Roland, Silver Ridge Lodge, 41°27’ S / 146°15’ E, 320 m, 25-28.I.2006, leg. M. Sommerer, BC ZSM Lep 0 8033. coll. ANIC (153, 16Ƥ): 13 ‘Mt Wellington 870m [Tas.] 6 Dec. 1980 L. Hill’; 13 ‘ 43.03S 146.17E Huon Camping Area Tas. 25 Jan 1983 J.C. Cardale’; 13 ‘Lake Fenton 1060m Tas. 14 Jan 1978 P.B. McQuillan’ ‘ ANIC Genitalia slide 18601’; 13 ‘Mt Read nr Rosebery Tas. 4–5 Dec 1990 P.B. McQuillan’; 13 ‘ 41.51S 145.33E Mt Read Tas. 500m 9 Jan 1989 P.B. McQuillan E.S. Nielsen’; 13 ‘ 43.25S 146.09E Melaleuca Tas. 15 Jan 1991 E.S. Nielsen E.D. Edwards’; 23 ‘ 43.25S 146.09E Melaleuca Tas. 28 Nov 1991 E.S. Nielsen M. Horak’; 13 ‘ 43.25S 146.09E Melaleuca Tas. 25 Nov 1991 E.S. Nielsen M. Horak’; 13 ‘ 43.25S 146.09E Melaleuca Tas. 14 Jan 1991 E.S. Nielsen E.D. Edwards’; 13 ‘ 43.25S 146.09E Melaleuca Tas. 17 Jan 1991 E.S. Nielsen E.D. Edwards’; 13 ‘ 43.23S 146.08E Claytons Bathurst Harb. Tas. 16 Jan 1991 E.S. Nielsen E.D. Edwards’; 1Ƥ ‘Mt Wellington 1000m [Tas.] 8 Dec 1980 L. Hill’; 1Ƥ “Mt Wellington 870m [Tas.] 6 Dec 1980 L. Hill’; 1Ƥ “ 42.02S 146.33E 12km NNE Bronte Park Tas. 20 Jan 1983 J.C. Cardale’; 1Ƥ ‘ 42.02S 146.33E 12km NNE Bronte Park Tas. 2 Feb 1983 J.C. Cardale’; 2Ƥ “Pensford Tas. 920m [Tas.] 22 Dec 1981 L. Hill’; 1Ƥ ‘ 40.58S 148.01E 1km SSE Gladstone Tas. 29 Jan 1983 J.C. Cardale’; 1Ƥ ‘Mt Nelson Tas. 160m 3 Nov 1979 P.B. McQuillan’; 1Ƥ ‘ 41.51S 145.33E Mt Read 500m Tas. 9 Jan 1989 P.B. McQuillan E.S. Nielsen’; 1Ƥ ‘ 43.25S 146.09E Melaleuca Tas. 14 Jan 1991 E.S. Nielsen E.D. Edwards’; 1Ƥ ‘ 43.23S 146.08E Claytons Bathurst Harb. Tas. 16 Jan 1991 E.S. Nielsen E.D. Edwards’; coll. TMAG: 13 ‘Tas. Pt. Sorell. Tas. U. V. L. 14.x.1988 L. Hill’ ‘Registration No. F4702’ ‘ TMAG genitalia slide F4703’; 13 ‘ 41°52’S 146°30’E TAS. Central Plateau L. Augusta to L. Ada 1150 m 3 FEB. 1994. P. B. McQuillan uvl’ ‘Registration No. F4704’ ‘ TMAG genitalia slide No. F4705’; 13 ‘ 41°52’S 146°30’E TAS. Central Plateau L. Augusta to L. Ada 1150 m 3 FEB. 1994. P. B. McQuillan uvl’ ‘Registration No. F4706’ ‘‘ TMAG wing slide No. F4707’; 13 ‘S.W. TAS. 12 Trees Hill (Strathgordon) U.V.L. 17.xi.1989. T. Semmens’ ‘Registration No. F4708’ ‘ TMAG genitalia slide F4709’; 13 ‘40°6’8.53”, 148°0’15.07” Bluff Rd. Whitemark Flinders Is. Tas. 31 Oct. 2008 C. J. Young U.V.L’. ‘Registration No. F4710’; 1Ƥ ‘41°11’5”, 146°19’5” Stony Rise Centre, Devonport. Tas. 23–29 Sept. 2004 L. Hill’; coll. ZSM: 1Ƥ S.W. TAS. 12 Trees Hill (Strathgordon) U.V.L. 17.xi.1989. T. Semmens’ ‘Accession No. 104062’; 1Ƥ ‘Mt Wellington 870m [Tas.] 12V UV 3 Nov. 1981 L. Hill’; 23 ‘40°6’8.53”, 148°0’15.07” Bluff Rd. Whitemark Flinders Is. Tas. 31 Oct. 2008 C. J. Young U.V.L’. ‘Accession Nos 104058, 104060’.

Other material examined: New South Wales, coll. ANIC (all excluded from the type series): 1Ƥ ‘Newnes S[tate].F[orest]. [ NSW] 20 Nov 1993 J.C. Keast’; 1Ƥ ‘ 36.28S 148.27E Rangers Stn 6km NE by E Thredbo NSW 1260m 6 Jan 2002 E.D. Edwards’; 1Ƥ ‘ 36.23S 148.25E 1.5km NNW Smiggin Holes NSW 1700m 9 Jan 2002 E.D. Edwards’; 1Ƥ ‘ 36.23S 148.25E Saddle 2km NW Smiggin Holes Kosciusko Nat. Pk 1680m 23 Jan 1987 E.D. Edwards’; 1Ƥ ‘ 1.3 km E of Island Bend NSW 1158m 30 Nov 1970 I.F.B. Common J.S. Dugdale’; 23 ‘ 36.23S 148.22E Guthega Village Kosciusko N.P. 1700m [ NSW] 24 Jan 1987 E.D. Edwards’, one with ‘ ANIC genitalia slide 18603’; 13 ‘ 36.23S 148.25E Saddle 2 km NW Smiggin Holes Kosciusko N.P. 1680m [ NSW] 23 Jan 1987 E.D. Edwards’.

Description. Wingspan 3 42–47 mm. Habitus and external characters ( Figs. 2–3 ): Forewing apex tapered, termen convex. Hindwing termen straight, angled at apex and tornus. Ground colour variable, usually purplish with dark suffusion and with blackish dotting of forewing costa. Fringe crimson. Antemedial line of forewing vague, at costa usually well marked by a costal spot. Postmedial line of forewing almost straight but slightly undulating, ochre with dark grey dots basally, obliquely leading into forewing apex. Border of postmedial line on the hindwing more violet and forked towards costa. Forewing apex with blackish fringe. Cell spots slightly elongate, black, with small hyaline filling. Underside of forewing with a large, round dark violet spot close to the inner termen at 2/3, i.e. at the position of the dotted postmedial line. Underside of hindwing with large white and grey speckled spot close to the costa at 2/3, i.e. at the position of the dotted postmedial line. Palpi, frons, and vertex concolorous with ground colour. Frons flat. Palpi at upperside of tip with dark grey scales, last segment narrow, length of palpi ca 1.5 times diameter of eye in both sexes. Proboscis well developed. Antennal flagellum thick in both sexes, male antennae unipectinate to 2/3 of length, female antennae filiform. Frenulum strong in 3, absent from Ƥ. Wing venation, forewing: R separate from Rs, anastomosing with Sc for a short distance in the forewing, R2–5 stalked. Hindwing: M2 located at mid-point between M1 and M3. Hindleg with four very short spurs in both sexes. Ansa narrow at the base, widening mesally, tapering apically. Last abdominal segment with extremely large interior coremata. 3 genitalia ( Fig. 8 ): Uncus long, narrow. Gnathos slender; medial process broad, sub-acute posteriorly, posterior surface covered with rows of short, broad spicules. Juxta large, well-sclerotised, divided. Saccus broad rectangular, with central invagination. Costa of valva sclerotised, setose. Valva asymmetrically adorned with subapical processes on ventral margin: left valva with single, large, sclerotised subapical, flattened, acute process, right valva with two short, broad processes. Aedeagus comparatively broad; vesica well sclerotised posteriorly, with longitudinal ridges; long narrow sclerotised process attached anteriorly, no discrete cornuti; caecum long, slender, tapered. Ƥ genitalia ( Fig. 10 ): Apophyses anteriores, posteriores long, slender. Lamella postvaginalis membranous. Lamella antevaginalis poorly developed. Sternum A7 slightly sclerotised. Ductus bursae short, close to corpus bursae strongly sclerotised, towards antrum dilating. Corpus bursae with strong anterior and posterior dilatations, constricted between; posterior half sclerotised, strongly ridged; anterior half membranous, plicate. Signum absent.

Morphological diagnosis (most congeners illustrated in BOLD (2008)): Oenochroma vinaria in habitus and external characters ( Figs. 4 , 6 ) very close to Oenochroma barcodificata and externally only distinguished, usually, by the straighter postmedian fascia of the forewing, often with a continuous dark proximal border. Dark suffusion of ground colour and blackish dotting of forewing costa usually reduced. Oenochroma pallida Warren, 1898 differs from both Oenochroma barcodificata and Oenochroma vinaria at once by the fore tibiae having an anterior apical hook (cf. Prout 1910, Turner 1932), and by the ochreous brown fringe of wing termen, the larger forewing cell spot on a paler ground colour, inner termen of hindwing underside with a narrow, dark spot; Oenochroma orthodesma Lower, 1894 has pale ochreous grey ground colour and ochreous fringe, an ochreous postmedian line edged anteriorly with pale yellow, the (ochreous) discal dots mostly wanting, hindwing at apex with pink suffusion and no spot on underside; Oenochroma decolorata Warren, 1896 , has grey forewings with fine brown irroration and purplish fringe, and a pale ferruginous postmedial fascia on both wings; Oenochroma phyllomorpha Lower, 1899 is of light brown ground colour, the forewing postmedial line is sinuate, fringe fuscous, and the cell spot lacking; Oenochroma cycnoptera Lower, 1894 has anterior tibiae with a strong apical hook (cf. Turner 1932), a very faintly ochreous postmedian line not reaching apex of forewing, fringe pale brownish, hindwings pale, without pattern and with whitish fringe; the New Caledonian Oenochroma unifasciata Holloway, 1979 , is broad-winged, without discal dots to the forewing.

Molecular diagnosis (see Fig. 1 and Table 2): DNA barcode analysis revealed a 3.34% K2P divergence between Oenochroma vinaria and O. barcodificata . Each species displays a very low mean intraspecific variation, with 0.05% (SE=0.03%, maximum distance of 0.33%) and 0.03% (SE=0.03%, maximum distance of 0.15%) for O. vinaria and O. barcodificata respectively and are thus unambiguously characterized by their DNA barcodes.

Distribution. Recorded in south-eastern and northern parts of Tasmania at altitudes from 0 to 870 m above sea-level, probably distributed all over the island. Further material from the central and southern tablelands of New South Wales dissected (T Edwards) but excluded from the type series to maintain geographical homogeneity in the type material.

O. barcodificata 0.03% 0.03% 0.15% n=9

O. pallida 0.00% n=3

O. decolorata 0.15% n=2

continued.

B mean distance standard error minimal distance O. vinaria vs O. barcodificata 3.34% 0.65% 3.1% O. barcodificata vs O. pallida 5.35% 1.17% 5.2% O. barcodificata vs O. decolorata 6.31% 1.02% 6.2%

Biology and morphology of immature stages.

Egg: Marked on all surfaces by round to hexagonal, concave cells with broad walls ( Figs 13, 15–18 ), micropylar cell walls narrow ( Fig. 14 ). Aeropyles, slightly elevated, openings very small ( Fig. 16 ), present on all surfaces apart from top of wider lateral side ( Fig. 17 ). Micropylar cells slightly recessed. Micropyles offcentred ( Fig. 13 ). Chorion undulating, irregularly pitted ( Figs 16–18 ). Colour: Off-white ( Fig. 12 ), becoming irregularly blotched rust-red, then transparent grey on maturity. Size (mm): (n= 4) L = 1.01 ± 0.01 (SE), W = 0.82 ± 0.01 (SE), T = 0.69 ± 0.01 (SE). Width/length: 0.81. Aeropylar opening size (micrometers) (n=10): L = 1.44 ± 0.07, W = 1.08 ± 0.04; relative aeropylar opening size [aeropyle length (micrometers)/egg length (mm)]: 1.4. Micropyles ( Figs 13, 14 ): distinct; no. of openings: 6; no. of cells in rosette: 9; no. of rows of cells in micropylar area: 5. Shape: Broad, bluntly ovoid, dorso-ventrally flattened, anterior pole angled to horizontal axis ( Figs 12 , 13, 15 ).

Oviposition: 2Ƥ code nos.: G87, G307. Oviposition period: 26 Sept.–31 Oct. Batch size range: 5–60 (n = 2). Realised fecundity (range): 4–60. Incubation time (days): 9. Batch configuration and attachment: Attached, singly.

Larval description. First instar (newly emerged): Head capsule width: 0.53 ± 0.03 mm (SE) (n=4). Head ground colour burnt yellow with chevroned dark-brown blotches, mouthparts lighter in colour, stemmata black, frons convex. Thorax and abdomen ground colour cream, except A6-A10 yellow, with numerous longitudinal wavy, dark brown stripes; posterior third of each segment, except A6-A10 and thoracic segments white; thoracic stripes less dense; venter dark chocolate brown. Thorax and abdomen becoming a uniform chocolate brown broken by numerous thin, wavy cream stripes after feeding. Setae very short, white, blunt on small dark brown pinacula on small raised black tubercles. Thoracic legs white with sparse dark brown spots. Prolegs on A5, A6, A10, rudimentary on A5. Spiracles very small, brown. Anal shield, no stripes, yellow with sparse orange streaks. Crochet arrangement, incompletely interrupted mesoseries. Resting position about 30° from substrate.

Second instar: Head capsule width: 0.91 ± 0.01 mm (SE) (n=7). Head ground colour light yellow with dark-brown blotches, dorso-lateral orange stripes, mouthparts pale brown, stemmata black, frons convex. Thorax and abdomen ground colour dark brown, broken by numerous longitudinal, narrow wavy, white stripes, venter smoky brown. A1 with dark brown spot on dorso-lateral line adjacent to anterior margin, directly adjacent and anterior to burnt orange blotch directly ventral to mid-dorsal line; A3 with two small orange blotches directly adjacent to mid-dorsal line, about one third segment length from posterior margin in some specimens, venter with black speckles. Setae very short, black, blunt on small dark brown pinacula on small raised black tubercles. Thoracic legs white with small brown blotches. Prolegs on A5, A6, A10, rudimentary on A5, same ground colour as body. Spiracles small, cream, peritremes black. Anal shield, same ground colour as body.

Third instar: Head capsule width: 1.48 ± 0.03 mm (SE) (n=5). Head ground colour cream with chevroned black blotches dorsally just lateral of mid-dorsal line, becoming two large circular, black blotches with orange centres, just posterior of frontoclypeus, dorso-lateral orange stripes, frontoclypeus pale brown streaked with black and brown, other mouthparts pale brown; stemmata black, frons convex. Thorax and abdomen ground colour cream, broad, dense-grey mid-dorsal band, centred by narrow white stripe connecting mid-segmental diamond shapes, thin wavy, grey stripes on rest of dorsum becoming yellowish-brown laterally. A1 with medial horn-like protuberances just lateral of mid-dorsal line, apex with black pinacula and setae, colour burnt orange with diagonal yellow stripe extending from mid-dorsal line to apex, black velvety blotch just anterior of protuberance at dorso-lateral position; similar small protuberances on A3. A8 D1 pinacula large, raised, bright burnt orange, venter paler from A3 to A10. Setae short, black, blunt, longer, transparent on venter A6- 10. Thoracic legs cream with small black blotches. Prolegs on A5, A6, A10, rudimentary on A5, same ground colour as body. Spiracles pale brown, peritremes black. Anal shield, same ground colour as body.

Fourth instar: Head capsule width: 2.31 ± 0.04 mm (SE) (n=4). Head ground colour light orange-brown mottled heavily with dark orange-brown, lighter stripes extend from mid-dorsal line to frons, becoming lighter and wider anteriorly, mouthparts pale brown, stemmata black, antennae brown; frons convex. Thorax and abdomen ground colour light brown, scattered sparsely with small white spots; dorsal band white speckled with black, on thoracic segments band is defined loosely by lines of black spots, on abdominal segments band is constricted at segment margins and bulges at segment centres; large fleshy horn-like dorsolateral protuberances on A1 just posterior of middle of segment, protuberances on A2 similar to A1 but smaller, apices of protuberances mottled orange-brown, large orange blotch mottled yellow, just anterior of protuberances, not extending laterally ventral of protuberances; venter pale brown speckled black; D1 pinacula on A8 on bright orange protuberances. Thoracic legs pale brown with sparse small black blotches. Prolegs on A5, A6, A10, rudimentary on A5, same ground colour as body. Spiracles orange, peritremes black, on white blotched with grey large spot.

Fifth instar ( Fig. 19 ): Head capsule width: 3.78 ± 0.03 mm (SE) (n=4). Length: 45–50 mm (n=3). Similar to 4th instar, except for the following. Head ground colour pinkish brown with faint orange marbling, mouthparts pale brown, stemmata black, antennae reddish brown; frons convex. Thorax and abdomen ground colour light brown, scattered sparsely with small white spots; dorsal stripe light brown speckled with small black spots, two lines of small white spots circled in black define dorsal stripe on abdominal segments. Two large fleshy dorso-lateral protuberances on A1, apices black anteriorly, yellow posteriorly, horns preceded by large mid-dorsal yellow blotch narrowing anteriorly to A1 anterior margin; relatively smaller protuberance on A3 similar to protuberance on A1. Venter pale brown blotched dark brown, white between A6 –A10. D1 pinaculi on A8 on bright orange protuberances. Black spots surround pinaculi of sub-dorsal setae. Setae short, pale brown, blunt. Thoracic legs pale brown with sparse small black blotches. Prolegs on A5, A6, A10, rudimentary on A5, same ground colour as body. Spiracles orange, peritremes black. Chaetotaxy: trisetose SV setae on A1; SV1, SV3 and V1 setae on A1 unaligned; three lateral setae on A6 proleg.

Pupa. Length: ɗ 23, 24 mm; Ψ, 24, 25 mm. Width: ɗ 7, 7 mm; Ψ, 6, 8 mm. Colour: reddish-brown on maturity; wings duller than shiny abdomen. Silken cocoon constructed from soil, debris and body fluids. Pupa large, stout. Labrum sub-trapezoidal, slightly convex, well-defined. “Mandibles” flat, rugose, margin between labrum and “mandibles” broadly ridged. Labial palpus large, pentagonal, rugose. Border between genae, maxilla less steep than that between oculus and pro-leg; border between pro-tibia and antenna about same length as border between pro-tibia and oculus. Pro-tibia same length as mid-tibia. Pro-femora not visible, Protibia meets maxilla at 2/3 length of maxilla, mid-tibia meets maxilla at 5/6 length of maxilla, antenna broad, almost reaches apex of maxilla, hind-leg barely visible. Hindwing becoming concealed at A4. Meta-notum short. Wing-bud callosity pronounced. Thoracic spiracles not visible, spiracles visible on A2-A8, poorly developed on A8; elliptical, elevated, pre-spiracular slit visible. Punctation on A1-A8, punctures small, shallow, numerous on A2-a7, sparse on A1, A8; randomly arranged, uniformly sparse on venter. Setae very short. Anal area large, anal slit bordered by longitudinal furrows. Male genitalia area, simple longitudinal slit. Cremaster, long, slender, roughly trigonal, posterior third, very rugose; one pair of terminal, robust, hamate, long, parallel setae, directed ventrally; pronounced dorsal and lateral grooves, six pubescent teeth on anterior margin of A10.

Larval hostplants. Reared on Grevillea sp. ( Proteaceae ).

Etymology. The species name refers to the barcoding campaign for Australia, and especially the fact that the new species could be distinguished by DNA barcoding without the need for dissecting (and thus damaging) an antique type specimen.

Figures

FIGURE 1. Neighbor joining tree for 34 Australian specimens in the genus Oenochroma (Kimura 2 Parameter, built with MEGA 4; all codon positions unweighted) based on sequences of the mtDNA COI gene (barcoding fragment 5 ’). Values above branches are bootstrap support values superior to 95 %. Terminals are identified by their process ID code on BOLD.

FIGURES 8. Male genitalia. Oenochroma barcodificata sp. nov., Tasmania (paratype TASAG Accession No. 104048) (photo CY). Scale bars 2 mm.

FIGURE 12. Egg of Oenochroma barcodificata. sp. nov. freshly laid egg, scale bar 1 mm. (photo CY)

FIGURE 19. Oenochroma barcodificata sp. nov. mature larva on Grevillea sp. Scale bar 10 mm (photo CY)

FIGURES 2, 3. Oenochroma barcodificata sp. nov., Ƥ holotype, Tasmania. 2: dorsal view. 3: labels (photos AH). Scale bar 2 cm.

FIGURES 10, 11. Female genitalia. 10: Oenochroma barcodificata sp. nov., Tasmania (paratype, DNA barcode LOTSA 125 - 06 / 06 - TASA- 00125) (photo CY). 11: Oenochroma vinaria, South Australia (gen. prp. ZSM G 13864) (photo AH). Scale bars 2 mm.

FIGURES 4, 5. Oenochroma vinaria, Ƥ lectotype of Guenée (photos RR / AM). 4: dorsal view. 5: labels (photos AM, RR). Scale bar 2 cm.

FIGURES 6, 7. Oenochroma vinaria, Ƥ lectotype of Monoctenia decora Wlk. 6: dorsal view. 7: labels (photos Peter Marriott and Peter Lilywhite, MVMA, Melbourne, Australia).

FIGURES 13 – 18. Egg of Oenochroma barcodificata. sp. nov. 13: anterior pole, scale bar 300 micrometers. 14: micropylar area, scale bar 200 micrometers. 15: whole egg, scale bar 1 mm. 16: aeropyles, scale bar 35 micrometers. 17: chorion on middle top of egg, scale bar 50 micrometers. 18: lateral chorion, scale bar 500 Μm. (photos CY).