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Zootaxa 1759: 43–50 (2008) www.mapress.com / zootaxa/

ISSN 1175-5326 (print edition)

Copyright © 2008 · Magnolia Press

ISSN 1175-5334 (online edition)

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A new species of Philophylla Rondani (Diptera: Tephritidae: Trypetini) from New Caledonia, recognized based on female postabdominal structure and molecular sequence data HO-YEON HAN1 & ALLEN L. NORRBOM2 1

Department of Life Science, Yonsei University, Wonju-si, Gangwon-do, Korea.E-mail: [email protected] Systematic Entomology Laboratory, PSI, ARS, USDA, c/o National Museum of Natural History, MRC-168, Washington, DC, U.S.A.Email: [email protected] 2

Abstract Philophylla millei, n. sp., from New Caledonia is described and its relationship analyzed. This species was recorded previously as Anastrephoides sp. based on a single female, which closely resembles the eastern Palaearctic species Anastrephoides matsumurai Shiraki. Whether this similarity reflects the close relationship of these species or a case of convergent evolution was examined using morphology and molecular data. We examined both male and female specimens of the New Caledonian trypetine species and sequenced the mitochondrial 16S ribosomal RNA gene of this species and related trypetine species. This new species is a member of the genus Philophylla Rondani based both on the female postabdominal structure as well as DNA sequence data. Key words: Diptera, Tephritidae, Trypetini, Philophylla millei, New Caledonia, 16S

Introduction The fruit fly fauna of New Caledonia was reviewed by Norrbom and Hancock (2004) who listed 25 species including three new species and six species newly reported from the island. Almost half the listed species are known only from New Caledonia, showing a high level of endemism. Norrbom and Hancock (2004) reported the only New Caledonian species of the tribe Trypetini as Anastrephoides sp. based on a single female, which closely resembles the eastern Palaearctic species Anastrephoides matsumurai Shiraki. We investigated whether this similarity reflects the close relationship of these two species or a case of convergent evolution. We clarify its generic classification, provide a detailed new species description, and discuss its phylogenetic position based both on morphological and molecular data.

Materials and methods The terminology and morphological interpretations used in this paper follow the glossary of White et al. (1999) and Han and Norrbom (2005). The type specimens of Philophylla millei are deposited in the Museum National d’Histoire Naturelle, National Collection of Insects, Paris, France. Voucher specimens of other species used in this study are deposited in Department of Life Science, Yonsei University, Wonju Campus, Korea. Molecular methods follow Han (2000) and Han and Ro (2005). We added the sequences of Anastrephoides matsumurai and five Philophylla species to the original 16S rDNA data set of Han (2000) to test the

Accepted by D. Bicklel: 5 Apr. 2008; published: 30 Apr. 2008

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phylogenetic position of the new species. As a result, 1289 bp long aligned sequences were assembled for 30 species and the data were analyzed using the same analytical methods of Han (2000) for comparative purpose. Species, collection and voucher data, and GenBank accession numbers are presented in Table 1. The alignment is available as a Nexus file from the first author upon request.

TABLE 1. Collection and voucher information of the tephritid flies sequenced. Status of the voucher specimens, voucher specimen numbers, and GenBank accession numbers are indicated in parentheses. Data concerning other flies analyzed in this study are listed in Han (2000). Anastrephoides matsumurai Shiraki. KOREA: Gangwon-do, Wonju-si, Heungeop-myeon, Hwaechon, 1.VII.1996, H.Y. Han & H.W. Byun. (both wings of the sequenced specimen mounted on a rectangular paper; YSUW08001; EU010373). Philophylla pulla (Ito). KOREA: Gyeongsangnam-do, Geoje-si, Mt. Nojasan, 4-5.VI.1997, D.S. Ku (both wings of the sequenced specimen mounted on a rectangular paper; YSUW08002; EU010374). Philophylla caesio (Harris) SWITZERLAND: SG 400m, Attenrhein, 4.VIII.1987, B. Merz (both wings of the sequenced specimen mounted on a rectangular paper; YSUW08003; EU010375). Philophylla marumoi (Miyake). KOREA: Gangwon-do, Inje-gun, Girin-myeon, Mt. Jeombongsan, 4.VIII.2006, D.S. Choi et al. (DNA extracted from the abdomen without destroying cuticular structures; the detached abdomen placed in a genitalia vial with the pinned voucher specimen; YSUW08004; EU010376). Philophylla fossata (Fabricius). KOREA: Gangwon-do, Pyeongchang-gun, Yongpyeong-myeon, Mt. Gyaebangsan, south valley, 10.VIII.1996, H.Y. Han & H.W. Byun. (Same specimen without one leg; YSUW08005; EU010377). Philophylla millei Han and Norrbom, n. sp. NEW CALEDONIA (the holotype—see Type Material; YSUW08006; EU010378).

Philophylla millei Han and Norrbom, n. sp. (Figs 1–15) Anastrephoides sp. Norrbom and Hancock, 2004: 67 (in New Caledonian list).

Type material. HOLOTYPE male: NEW CALEDONIA, Sarraméa Col d’Amieu, Malaise trap, 8.III.2006, C. Mille (the genitalia dissected and kept in a genitalia vial; left mid and hind legs removed for DNA extraction). PARATYPE female, 9.7 km NW Sarraméa, Malaise trap along Melaleuca Path, 15.I.1996, M.E. Irwin, D.W. Webb & E.I. Schlinger (the postabdomen dissected and kept in a genitalia vial). Both types are deposited in Museum National d’Histoire Naturelle, Paris. Description. Body (Fig. 1) almost entirely shiny to subshiny yellow brown with dark brown setae and setulae; wing length 3.9–4.2 mm; thorax length 1.9 mm. Head (Figs 2, 3) yellow brown with ocellar tubercle dark brown, frons-head ratio (narrowest width of frons in dorsal view / width of head) 0.28, eye ratio (shortest eye diameter / longest eye diameter) 0.72–0.74, gena-eye ratio (genal height between ventral eye margin and ventral genal margin anterior to genal seta / longest eye diameter; gena measured with head tilted slightly dorsally so that gena is at its broadest) 0.08–0.10, arista-antenna ratio (length of arista / length of antenna excluding arista) 1.9–2.1; frons with 2 orbital and 3 evenly spaced frontal setae; medial vertical seta 0.7–0.8x as long as longest diameter of eye; lateral vertical seta 0.6x as long as medial vertical seta; postocellar seta 0.4x as long as medial vertical seta; paravertical seta hair-like, about 0.3x as long as medial vertical seta; ocellar seta 2.5x as long as ocellar tubercle; scape and pedicel yellow brown with short dark brown setulae; flagellomere 1 yellow brown; arista short pubescent, dark brown except for yellow brown basal thickened part; face more or less flat; parafacial narrow, 0.2x as wide as flagellomere 1; facial ridge with fine dark brown setulae; genal seta strong (Figs 2, 3); postgena moderately swollen with relatively long dark brown setulae; postgenal seta strong; occiput flat, yellow brown; postocular setae extended 0.6x distance from upper

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eye margin to lower eye margin; supracervical setae dark brown; mouthparts short with labella yellow brown setulose and palpi dark brown setulose. Thorax entirely yellow brown with dark brown setae and setulae; scutum subshiny yellow brown, slightly microtrichose; 1 postpronotal, 2 scapular, 1 acrostichal, 1 dorsocentral, 1 intra-alar, 1 presutural supra-alar, 1 postsutural supra-alar, 1 postalar and 2 notopleural setae; dorsocentral seta slightly posterior to level of postsutural supra-alar seta; scutellum flat with sparse fine dark brown marginal setulae; basal scutellar seta 2.5x as long as scutellum; apical seta 1.7x as long as scutellum, crossing subapically; proepisternum densely covered with long dark brown setulae; ventral anepisternal seta slightly shorter than dorsal one; katepisternal seta well developed; mediotergite shiny yellow brown. Legs entirely yellow brown with dark brown setae and setulae; fore femur with 7 posteroventral setae; fore tarsomere 1 as long as remaining tarsomeres together; tibial spur 1.8x as long as wide; hind femur with 2 subapical dorsal setae; hind tibia with row of short anterodorsal setae. Wing (Figs 1, 4) hyaline with orange brown to brown pattern, including: subbasal band from crossvein h to apex of cell bcu, cell bc and extreme base of cell br yellow, cell c brown on basal 1/3 and apical 1/5; brown subcostal band from pterostigma to BM-Cu, covering fork of Rs, becoming orange brown in cells br and dm where connected to radial-medial band so that apical half of br lacks hyaline area, also connected to subbasal band in cells br and bm; orange brown radial-medial band extending obliquely from costal margin in middle of cell r1, covering R-M, and turning posteriorly to reach wing margin in cell cu1 slightly distal to vein A1+Cu2, connected to orange brown to brown anterior apical band in cell r1 and to subcostal band in cells br and dm to form S-shaped mark; orange brown to brown posterior apical and subapical bands broadly connected to form inverted V-shaped mark, subapical band also broadly connected posteriorly to radial-medial band but separated anteriorly from anterior apical band. Wingthorax ratio (wing length / thorax length) 2.1–2.3, vein R4+5 ratio (distance along vein R4+5 between crossvein R-M and vein R4+5 apex / distance between crossvein R-M and basal node of vein R4+5) 2.9–3.0, vein M ratio (distance along vein M between crossveins R-M and DM-Cu / distance between crossveins R-M and BM-Cu) 1.0–1.1, subcosta-costa ratio (length of pterostigma / length of costal cell, both measured along vein C) 0.29– 0.32; R4+5 dorsally with 13–14 tiny setulae between node and R-M, 9–15 setulae apical to R-M; cell bm, cell bcu, anal lobe, and alula, except along fold, entirely microtrichose; posteroapical extension of cell bcu about half as narrow as width of wing veins, about 1.5x as long as crossvein BM-Cu. Male abdomen subshiny yellow brown with dark brown setae and setulae, slightly longer than wide; epandrium (Figs 10, 11) yellow brown with yellow brown to brown setae and setulae; lateral surstylus yellow brown with more or less pointed apex in profile (Fig. 11), anterior lobe modified into prensiseta-like structure; medial surstylus with single prensiseta; prensiseta together with modified anterior lobe of lateral surstylus superficially appear as two prensisetae (Fig. 10); proctiger pale yellow with yellow brown setulae; medial sclerite of glans (Fig. 12) without internal pattern of granulation; dorsal sclerite of glans (Fig. 12) with faint internal sculpture pattern of narrowly rhombic cells dorsally. Female abdomen entirely subshiny yellow brown with dark brown setae and setulae; preabdomen slightly longer than wide; oviscape (Fig. 13) slightly longer than wide, entirely yellow brown, dorsally with a pair of strong lateral marginal setae and ventrally with a pair of moderate sublateral marginal setae; eversible membrane medially with strong triangular denticles and posteriorly with smaller denticles; dorsal and ventral taeniae about 0.3x as long as total length of membrane; aculeus (Fig. 15) broad, parallel-sided with apical onethird tapered; tip triangular, with moderately large lateral serrations; medial membrane under ventral plates (= sternite 8) almost entirely covered with numerous anteriorly directed denticles; 3 spermathecae (Fig. 14) similar in size, brown, about twice as long as wide with numerous transerverse furrows; short apical portion of spermathecal duct yellowish brown. Etymology. This species is named for Mr. Christian Mille, who acted upon our request to search for more specimens of this rare species. He ran traps in the area where the female had been found and collected the only known male (the holotype) which he promptly shipped so that DNA extraction as well as genitalic examination was possible. A NEW SPECIES OF PHILOPHYLLA

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FIGURES 1–9. 1–3. Philophylla millei, n. sp., holotype male. 4. P. millei, n. sp., paratype female. 5. Anastrephoides matsumurai Shiraki, female. 6. Myoleja korneyevi Han & Kütük, female. 7. M. korneyevi, male. 8. Anastrepha barnesi Aldrich. 9. Anastrepha obliqua (Macquart). Abbreviations: AAB = anterior apical band; PAB = posterior apical band; RMB = radial-medial band; SAB = subapical band; SCB = subcostal band.

Remarks. Adding P. millei to Philophylla could cause taxonomic confusion because its wing pattern is aberrant for the genus. Therefore, we provide here the following modifications of the diagnosis previously published for Philophylla and the section of the key to the genera of Trypetini involving the genus. Generic diagnosis (modified from Han, 1999: 284): Philophylla can be defined as a monophyletic group by the following unequivocal synapomorphies: (1) oviscape dorsally with a pair of strong lateral marginal setae (Han, 1999, fig. 11.9A); and (2) lateral surstylus with posterior lobe elongated and anterior lobe broadly flattened (all species for which male genitalia have been examined except P. millei) (Han, 1999, figs 11.6A, B) or modified into a prensiseta-like structure (P. millei; this state here interpreted as secondarily derived). Although dissection of genitalia is recommended to confirm generic placement, most species can be conveniently recognized as Philophylla by having one of four typical wing patterns (Fig. 4; Han 1999, figs 11.5D– F).


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FIGURE 10–15. Philophylla millei, sp.n.; 10. epandrium and surstyli, posterior view (proctiger removed); 11. epandrium, surstyli, and proctiger, lateral view; 12. glans, dorsolateral view (inset at 8x main figure); 13. female postabdomen, ventral and dorsal views (insets at 8x main figures); 14. spermatheca; 15. aculeus, ventral and dorsolateral views.

Philophylla millei superficially resembles Anastrephoides matsumurai Shiraki, Myoleja korneyevi Han and Kütük, and M. sinensis (Zia) of the tribe Trypetini, especially in wing pattern (Figs 4–7), but it can be distinguished from these species by its entirely orange brown cell br, which lacks the hyaline area present in the other species distal to the fork in vein Rs. Because wing patterns very similar to that shown in Han (1999, fig. A NEW SPECIES OF PHILOPHYLLA


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11.5F) occur also in some species of Anomoia, Hoplandromyia, and a few other genera, extra precaution is needed.

FIGURE 16. Relationships of the tribe Trypetini inferred from neighbor-joining tree based on Kimura two parameter distances (1159 bp after gaps and sites with missing data removed). The first number is the Pc value from the standard error test (higher than 90%), and the second number is the Pb from the bootstrap test (2000 replications).

Modification to Key to the Genera of Trypetini (Han, 1999: 275): 24.

Wing apically with two inverted L-shaped bands (Han, 1999, fig. 11.5.C)..........................................24a Wing apically without two inverted L-shaped bands..............................................................................25 24a. Wing cell br distal to fork in vein RS with large hyaline area (Han, 1999, fig. 11.5.C)............................. .............................................................................................................................. Anastrephoides Hendel Wing cell br almost entirely orange brown......................................Philophylla millei Han and Norrbom Characters of the male genitalia further separate this species from any other species of Trypetini: 1) medial surstylus with single prensiseta (Fig. 10); 2) anterior lobe of lateral surstylus modified into prensisetalike structure (Fig. 10); 3) acrophallus of glans without internal sculpture pattern of round granulation (Fig. 12); and 4) with two small round apical sclerites anterior to acrophallus (Fig. 12). The generic classification of this species is based on the female postabdominal structure as well as DNA sequence analysis (see Discussion).


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Discussion As mentioned in the Remarks, Philophylla millei resembles Anastrephoides and some Myoleja species of the tribe Trypetini in wing pattern. Anastrephoides Hendel belongs to the subtribe Trypetina, whereas Myoleja Rondani is classified in the subtribe Chetostomatina. This type of wing pattern (Figs 4–9; presence of both Sshaped and inverted V-shaped bands) also is found in most species of the Neotropical genus Anastrepha Schiner (tribe Toxotrypanini), although the location of the basal part of the band differs in the latter genus (it continues to cell bcu; Figs 8, 9). We believe that these wing patterns represent a case of convergent evolution. Philophylla millei was identified initially as a new species of Anastrephoides based on a single female (Norrbom and Hancock, 2004), which is very similar to A. matsumurai both in wing pattern (Figs 4, 5) as well as body coloration. However, dissection of its postabdomen showed that it possesses one of the two synapomorphies of the genus Philophylla (subtribe Trypetina) suggested by Han (1999) (Fig. 13; apex of oviscape dorsally with a pair of strong lateral marginal setae). Therefore, examination of a male specimen to confirm the other synapomorphy became necessary to clarify the generic membership of the species. Due to the generous help of C. Mille, we obtained a single fresh male (the holotype). However, the male genitalia turned out to be highly apomorphic not only for the genus Philophylla but also for the subtribe Trypetina. Instead of possessing the male synapomorphy of Philophylla (lateral surstylus with the anterior lobe broadly flattened and the posterior lobe elongated), P. millei has its anterior lobe modified into a prensisetalike structure (Fig. 10). Because this species has a single prensiseta (almost all the species of Trypetini have two), the anterior lobe might supplement the function of the prensiseta. Furthermore, the glans (Fig. 12) lacks the synapomorphy of the subtribe Trypetina (medial sclerite with internal sculpture pattern of round granulation). In short, examination of the male genitalia provided no additional evidence concerning its generic placement. Fortunately we were able to extract DNA from the two legs detached from the holotype and sequence the mitochondrial 16S ribosomal RNA gene. We sequenced four additional Philophylla species as well as Anastrephoides matsumurai, and incorporated them into the existing molecular dataset by Han (2000). The resulting inferred phylogenetic tree clearly showed that this species belongs to the genus Philophylla (Fig. 16). All five selected Philophylla species, including the type species P. caesio (Harris), form a strong monophyletic group with extremely high statistical support (standard error test value (Pc) / bootstrap test value (Pb) = 99/ 99). Among these species, P. millei shows the closest relationship to P. fossata (Fabricius). On the other hand, the superficially similar A. matsumurai evidently belongs to the Trypeta genera group (sensu Han, 1999) within the subtribe Trypetina. We interpret these results to indicate that the shape of the anterior lobe of the lateral surstylus in P. millei is secondarily derived from the shape found in other species of Philophylla and the lack of internal sculpture on the medial sclerite of the glans in P. millei is due to secondary loss (homoplasy). Because of the small sampling of Philophylla species in our analysis, it is possible that the clade of P. millei and P. fossata shown in Fig. 16 might actually represent a larger monophyletic group. An additional analysis at least including a reasonable number of the Australasian Philophylla species is necessary to elucidate the phylogenetic position of P. millei. Han (1992, 1999, 2005) defined the genus Philophylla as a monophyletic group and resurrected the generic name from the synonymy of Myoleja. The 56 currently recognized species (updated from Han (1992) and Norrbom et al. (1999)) are distributed throughout the Old World with their highest diversity in the Oriental Region (14 Palaeactic, 28 Oriental, 15 Australasian (including Oceania), and 11 Afrotropical species). Six Australasian species inhabit the island chain from the Solomon Islands east to New Caledonia and Fiji. Among these species, P. fossata is the most widespread, occurring from India, Korea, and Japan southeast to the Solomon Islands. Another widespread species, P. conjuncta (Meijere), occurs from Indonesia to the Bismark Archipelago and Solomon Islands. On the other hand, P. propreincerta (Hardy) is endemic to the Solomon Islands, P. millei to New Caledonia, and P. bifida (Bezzi) and P. curvinervis (Bezzi) to Fiji. A more

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comprehensive species level phylogenetic study of Philophylla would provide an interesting insight into their historical biogeography. In this analysis (Fig. 16) the genus Philophylla was grouped with the subtribe Chetostomatina, suggesting that the subtribe Trypetina is paraphyletic. Because the statistical support for the associated branches are low, however, we are currently conducting a comprehensive molecular analysis based on many more trypetine taxa and four mitochondrial genes to clarify this problem as well as to further resolve generic relationships within the tribe Trypetini (Han and Ro, in prep.).

Acknowledgements We are grateful to Mr. Christian Mille (Institut Agronomique néo-Calédonien) for providing the freshly collected holotype specimen. Steve Gaimari and Don Webb loaned the paratype from material collected by Illinois Natural History Survey Staff. We also thank Drs. V.A. Korneyev (I.I. Schmalhausen Institute of Zoology, Ukraine) and K.E. Ro (Yonsei University, Korea) for kindly reviewing the manuscript. This study was supported by the Korean Ministry of Environment (the Eco-technopia 21 Project).

References Han, H.-Y. (1992) Classification of the tribe Trypetini (Diptera: Tephritidae: Trypetinae). Dissertation, Pennsylvania State University, University Park. Han, H.-Y. (1999) Phylogeny and behavior of flies in the tribe Trypetini (Trypetinae). In: Aluja, M. & Norrbom, A.L. (Eds), Fruit flies (Tephritidae): Phylogeny and Evolution of Behavior. CRC Press, Boca Raton, Florida, pp. 253– 297. Han, H.-Y. (2000) Molecular phylogenetic study of the tribe Trypetini (Diptera: Tephritidae), using mitochondrial 16S ribosomal DNA sequences. Biochemical Systematics and Ecology, 28, 501–513. Han, H.-Y. (2005) Prochetostoma, a new genus proposed as a basal group of the subtribe Chetostomatina (Diptera: Tephritidae). Biotaxonomy of Tephritoidea (ed. by A. Freidberg). Israel Journal of Entomology, 35–36, 147–162. Han, H.-Y. & Norrbom, A.L. (2005) A systematic revision of the New World species of Trypeta Meigen (Ditpera: Tephritidae). Systematic Entomology, 30, 208–247. Han, H.-Y. & Ro, K.-E. (2005) Molecular phylogeny of the superfamily Tephritoidea (Insecta: Diptera): new evidence from the mitochondrial 12S, 16S, and COII genes. Molecular Phylogenetics and Evolution, 34, 416–430. Norrbom, A.L. & Hancock, D.L. (2004) New species and new records of Tephritidae (Diptera) from New Caledonia. Bishop Museum Bulletin in Entomology, 12, 67–77. White, I.M., Headrick, D.H., Norrbom, A.L. & Carroll, L.E. (1999) Glossary. In: Aluja, M. & Norrbom, A.L. (Eds), Fruit flies (Tephritidae): Phylogeny and Evolution of Behavior. CRC Press, Boca Raton, Florida, pp. 881–924.


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