Systematic Entomology (2014), 39, 335–353

Signal through the noise? Phylogeny of the Tachinidae (Diptera) as inferred from morphological evidence

PIERFILIPPO CERRETTI1,2, JAMES E. O’HARA3, D. MONTGOMERY WOOD3, HIROSHI SHIMA4,DIEGOJ. INCLAN5 andJOHN O. STIREMAN III6

1Department of Biology and Biotechnology ‘Charles Darwin’, University of Rome ‘Sapienza’, Rome, Italy, 2Centro Nazionale Biodiversita` Forestale – Corpo Forestale dello Stato, Verona, Italy, 3Canadian National Collection of Insects, Agriculture and Agri-Food Canada, Ottawa, Canada, 4Kyushu University Museum, Kyushu University, Fukuoka, Japan, 5DAFNAE-Entomology, University of Padova, Padova, Italy and 6Department of Biological Sciences, Wright State University, Dayton, OH, U.S.A.

Abstract. The oestroid family Tachinidae represents one of the most diverse lineages of insect parasitoids. Despite their broad distribution, diversity and important role as biological control agents, the phylogeny of this family remains poorly known. Here, we review the history of tachinid systematics and present the first quantitative phylogenetic analysis of the family based on morphological data. Cladistic analyses were conducted using 135 morphological characters from 492 species belonging to 180 tachinid genera, including the four currently recognized subfamilies (Dexiinae, Exoristinae, Phasiinae, Tachininae) and all major tribes. We used characters of eggs, first-instar larvae and adults of both sexes. We examined the effects of implied weighting by reanalysing the data with varying concavity factors. Our analysis generally supports the subfamily groupings Dexiinae + Phasiinae and Tachininae + Exoristinae, with only the Exoristinae and the Phasiinae reconstructed as monophyletic assemblages under a wide range of weighting schemes. Under these conditions, the Dexiinae, which were previously considered a well-established monophyletic assemblage, are reconstructed as being paraphyletic with respect to the Phasiinae. The Tachininae are reconstructed as a paraphyletic grade from which the monophyletic Exoristinae arose. The Exoristinae are reconstructed as a monophyletic lineage, but phylogenetic relationships within the subfamily are largely unresolved. We further explored the evolution of oviposition strategy and found that the oviparous groups are nested within ovolarviparous assemblages, suggesting that ovipary may have evolved several times independently from ovolarviparous ancestors. This counterintuitive pattern is a novel hypothesis suggested by the results of this analysis. Finally, two major patterns emerge when considering host associations across our phylogeny under equal weights: (i) although more than 60% of tachinids are parasitoids of Lepidoptera larvae, none of the basal clades is unambiguously associated with Lepidoptera as a primitive condition, suggesting that tachinids were slow to colonize these hosts, but then radiated extensively on them; and (ii) there is general agreement between host use and monophyly of the major lineages.

Introduction

Correspondence: Pierfilippo Cerretti, Department of Biology and Within the order Diptera, the oestroid family Tachinidae re- Biotechnology ‘Charles Darwin’, University of Rome ‘Sapienza’, presents the largest lineage of endoparasitoids. Tachinid larvae Piazzale A. Moro 5, I-00185 Rome, Italy. E-mail: pierfilippocerretti@ yahoo.it develop within their hosts (mainly insects, but also chilopods

© 2014 The Royal Entomological Society 335 336 P. Cerretti et al. and scorpions) by feeding first on haemolymph and non- History of the classification of Tachinidae vital tissues and organs, and then on vital parts, with few exceptions killing their hosts before completing their larval The purpose of this section is to briefly review the development (Herting, 1960; Wood, 1987b; Mellini, 1991; history of tachinid classification and to consider some of Tschorsnig & Richter, 1998; Dindo, 2011). This finely tuned the morphological characters that have been used to define way of life is undoubtedly the most important ecological fundamental groups in recent classification schemes. For a trait that distinguishes these flies from other oestroids, more comprehensive and detailed account of the history of which are mostly sarcophagous, coprophagous, parasitic on tachinid classification, see O’Hara (2013b). vertebrates, or kleptoparasitic on Hymenoptera (with only a Coquillett (1897), in his revision of the Tachinidae of few being parasitoids, notably the Rhinophoridae and certain America north of Mexico, recognized the Tachinidae almost Sarcophagidae) (Feener & Brown, 1997). as it is known today but without the subfamily Dexiinae. At According to the recent phylogenetic reconstruction of about the same time, in Europe, Girschner (1893, 1896) was Wiegmann et al. (2011), cyclorrhaphan Diptera had an early elaborating on a system of chaetotaxy developed by Osten- Tertiary explosive radiation. Tachinids were probably radiat- Sacken (1881, 1884) and used the presence of ‘hypopleural’ ing rapidly by the mid-Tertiary, becoming widespread and (meral) setae to delimit the Tachinidae, a concept for the family abundant in virtually all terrestrial ecosystems (Wood, 1987b; equalling that of the present-day superfamily Oestroidea. This Tschorsnig & Richter, 1998). There are nearly 8200 named classification was largely followed in the influencial Katalog tachinid species (and thousands more undescribed), accounting der pal¨aarktischen Dipteren (Bezzi & Stein, 1907). Not until for about 6.5% of the total diversity of Diptera. Over 90% of the discovery of the value of the ‘metanotum’ (subscutellum; tachinid species attack phytophagous insects, and about 60% of character state 60:1 below) by Malloch (1923) did authors these are parasitoids of ditrysian Lepidoptera (Stireman et al., begin to use this character to delimit the Tachinidae along 2006; Cerretti, 2010), which comprise the most diverse group modern lines. of plant-feeding organisms. The progressive elements of Girschner (1893, 1896) were Given their broad distribution, great diversity and ecological further refined by Villeneuve (1924, 1933), whose work was, role as enemies of primarily herbivorous insects, tachinids in turn, further advanced by Mesnil (1939) in his Essai sur les have attracted considerable attention from basic and applied Tachinaires. researchers, as well as taxonomists (Stireman et al., 2006; Mesnil (1939) recognized six subfamilies of the Tachinidae: O’Hara, 2013b). However, the classification and identification Dexiinae, Phasiinae, ‘Larvaevorinae’ (Tachininae), Ameniinae of tachinids have posed formidable challenges, and these (currently in Calliphoridae), ‘Salmaciinae’ and ‘Phorocerinae’ have hindered the understanding of their biology, ecology and (the last two comprising present-day Exoristinae). He brought potential usefulness in controlling pests in managed systems. together in his Phorocerinae those tachinids possessing a haired The group is widely regarded as one of the more difficult prosternum (41:0) (i.e. character 41, state 0 in the present study, families of Diptera in terms of identification (Crosskey, 1976), see File S2) and a short and fine ‘prealar’ (first postsutural and a universal classification scheme remains elusive despite supra-alar) seta (49:1). The Phorocerinae were subdivided recent progress. into the ‘Phorocerini’ (Exoristini), Blondeliini and ‘Crocutini’ Here, as part of a larger initiative to understand the (Siphonini). Each was further characterized in part by: the phylogeny and evolution of the family Tachinidae on a global Phorocerini by vein M (as ‘4e’) having an angular bend (71:3) scale (Stireman et al., 2013), we present the first quantitative and shadow fold (72:1), and subapical scutellar setae divergent phylogenetic analysis of the family based on morphological (character not coded in analysis below); the Blondeliini by data. We briefly review the history of tachinid classification vein M having a rounded bend and no shadow fold (71:1), and and the hypothesized relationships that key workers have subapical scutellar setae divergent; and Crocutini by vein M advanced, noting some of the important character states having a rounded bend and no shadow fold (71:1, 2; 72:0), and that have been used to define various tachinid clades. We subapical scutellar setae convergent. The current concepts of then present an analysis of the phylogenetic relationships the Exoristini and Blondeliini largely date from Mesnil (1939). of the Tachinidae based on a broad range of taxa (180 The Exoristini are probably monophyletic (Stireman, 2002; genera) and a diverse collection of morphological characters Tachi & Shima, 2010), but the Blondeliini, according to Wood (135). Our aim is to assess the strength of the phylogenetic (1985), are probably not. Wood (1985: 7) noted that the small signal of morphology in defining major tachinid clades and prealar seta (49:1) ‘is probably a plesiomorphic character’. their relationships, despite the great degree of morphological Villeneuve’s contemporary in the New World was homoplasy in the family. We use our resulting phylogenetic Townsend, a prolific describer of tachinid genera and species. reconstruction to evaluate previous hypothesized relationships Townsend’s greatest achievement was his 12-volume Manual and classifications, and to propose novel ones that can of Myiology in which the genera of Tachinidae and allies be evaluated with subsequent morphological and molecular were diagnosed and assigned to a multitude of tribes and phylogenetic studies. Finally, we examine the implications families. This work was also Townsend’s greatest downfall, of our phylogenetic results for understanding evolutionary for unrelated genera were frequently grouped together into patterns of reproductive strategy and host use within the tribes. The catalogue of the Tachinidae of America south of family. the United States by Guimaraes˜ (1971) did not significantly

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 Phylogeny of Tachinidae 337 improve the classification of the region’s Tachinidae. The but reduced them in number and recognized 14 in the fauna of America north of Mexico has fared better in the Palaearctic Region. successive catalogues of Sabrosky & Arnaud (1965) and The treatments of Old World non-Palaearctic Tachinidae by O’Hara & Wood (2004), the latter much influenced by the Crosskey (1973, 1976, 1980, 1984) and Cantrell & Crosskey classification of Herting (1984). (1989) were conservative in their taxonomic approach, Mesnil dominated tachinid taxonomy in Europe for decades treating the Voriini in the Tachininae and the Neaerini and after his Essai, particularly through instalments to Die Siphonini in the Exoristinae, recognizing the Dufouriinae Fliegen der Palaearktischen Region (Mesnil, 1944–1975). as a subfamily, and not dividing the Goniini and Eryciini During this time, Herting published early works on the according to egg type. female postabdomen of calyptrate flies and the biology of The ‘gold standard’ of classifications remains that of Herting West Palaearctic Tachinidae (Herting, 1957, 1960). Implicit (1984), which was followed by Herting & Dely-Draskovits in these works was an attempt to order the Tachinidae (1993). Although this classification specifically applies to the phylogenetically. The Salmaciinae and Phorocerinae of Mesnil Palaearctic Region, its hierarchy of subfamilies and tribes (1939) [also Mesnil (1944–1956, 1956–1965) as Salmaciini provides a good framework for a global classification (Table 1), and Phorocerini] were restructured to form the near-modern at least at present (see the Discussion). Only portions of the Exoristinae with tribes Exoristini, Blondeliini, Acemyini (as classification are stable from a phylogenetic perspective, and ‘Acemyiini’), Siphonini, Ethillini, Winthemiini and Goniini. a greater understanding of tachinid relationships may indicate The Goniini were later split to become the modern Goniini where changes should be made to better reflect the evolutionary and Eryciini, with the former restricted to species producing history of the Tachinidae. microtype eggs (6:1) that are ingested by their hosts (9:1) Recently, Stireman (2002) and Tachi & Shima (2010) (Mesnil, 1975a,b; Herting, 1984). Thus constituted, the Goniini provided the first attempts at reconstructing phylogenetic are generally considered monophyletic; the Eryciini are relationships within the Tachinidae using molecular data. regarded as paraphyletic. The Siphonini were later moved to Stireman (2002) used two genes, EF1α and 28S rDNA, while the Tachininae (Herting, 1984). Tachi & Shima (2010) used four (16S, 18S, 28S rDNA and The modern concept of the Dexiinae, with the main tribes white). These studies focused on the subfamily Exoristinae, of Dexiini, Voriini and Dufouriini, dates from Herting (1957, generally supporting its monophyly and that of its constituent 1960). Herting used the as-yet-unpublished discoveries of tribes (with some exceptions). Notably, Stireman (2002) failed Verbeke (1962, 1963) to define the subfamily according to reconstruct a monophyletic Goniini (the microtype egg to unique features of the male terminalia: phallus with layers, state 6:1), while Tachi and Shima did find support a hinged connection between basiphallus and distiphallus for this clade. Both studies found a similar progression of (Verbeke’s ‘type II’ connection) (115:1) and pregonite strap- clades, with Winthemiini basal, then Exoristini branching off, like and connective (‘type C’ of Vebeke’s ‘posterior paramere’) followed by the Blondeliini and, finally, the Eryciini and (103:1). Goniini forming a clade. As suggested by this progression, In the first instalment on the ‘Larvaevorini oder Tachinini’ Tachi & Shima (2010) found, using character reconstruction, portion of Die Fliegen der Palaearktischen Region,Mesnil that ovolarviparous forms (state 7:1) have arisen repeatedly (1966) revised the classification of the tachinids he had dealt from oviparous ancestors (state 7:0). with in previous instalments and presented the classification he would follow for the Tachinini. The higher classification was summarized in a table (Mesnil, 1966: 882) wherein present-day Materials and methods Tachinidae were treated as a subfamily with six tribes: Voriini, Dexiini and Tachinini (on the left in the table) and Exoristini, Monophyly of terminal taxa Goniini and Phasiini (on the right in the table). Both Mesnil (1966) and Herting (1966) expressed the same views about the All terminal taxa included in the analysis are assumed, a two main branches of Tachinidae: priori, to be valid genera following the definitions given in all major monographs (Crosskey, 1984; Tschorsnig, 1985; Wood, (1) The three groups on the right in Mesnil’s table are 1987b; Tschorsnig & Herting, 1994; Tschorsnig & Richter, oviparous, or originally so, producing planoconvex eggs 1998; Cerretti, 2010; Cerretti et al., 2012a). We also assumed with a thick curved dorsal surface and a thin and flat that each species chosen for character coding is congeneric underside (state 2:1). with the type species of the genus in which it is placed. It is (2) The three groups on the left in Mesnil’s table are possible that some of these genera are in fact not monophyletic, ovolarviparous, producing eggs with a thin membrane and but assessing this is beyond the scope of this study. without differentiation into an upper and a lower surface (2:0). Selection of taxa, coding of character states and terminology Mesnil (1966) recognized the Tachinini as the most diverse group of tachinids and subdivided it into about 30 subtribes. Specimens belonging to 180 tachinid genera (total of 492 Herting (1984) treated these subtribes as tribes of Tachininae, species examined) were selected for character state coding

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 338 P. Cerretti et al.

Table 1. Subfamilial and tribal level classification of the Tachinidae, excluding tribes given in Table 2.

Dexinae Phasiinae Tachininae Exoristinae

Dexiini Catharosiini Bigonichetini Myiophasiini Anacamptomyiini Doleschallini Cylindromyiini Brachymerini Myiotrixini Blondeliini Dufouriinia Gymnosomatinib Ernestiinic Neaerini Eryciini Epigrimyiini Hermyini Germariini Nemoraeini Ethillini Rutiliini Leucostomatini Germariochaetini Occisorini Exoristini Sophiini Parerigonini Glaurocarini Ormiini Goniini Telothyriini Phasiini Graphogastrini Pelatachinini Thrixionini Uramyini Iceliini Polideini Winthemiini Voriini Leskiini Protohystriciini Macquartiini Siphonini Megaprosopini Tachinini Minthoini a Including Freraeini. b Including Trichopodini. c Including Linnaemyini and Loewiini. Taxa with at least one representative included in the present study are shown in bold.

(File S1). These taxa represent the four currently recognized Table 2. List of exemplar ‘enigmatic’ taxa and their subfamily tachinid subfamilies (Dexiinae, Exoristinae, Phasiinae, Tachin- placement by different authors. inae) and all of the major widespread tribes (Tables 1, 2). Blow Herting Tschorsnig O’Hara & Wood flies of the genera Calliphora Robineau-Desvoidy, Lucilia Tribe/genus 1984 1985 2004 Robineau-Desvoidy, Pollenia Robineau-Desvoidy, the rhiniid Rhyncomya Robineau-Desvoidy and the rhinophorids Phyto Imitomyiini PP D Robineau-Desvoidy and Rhinophora Robineau-Desvoidy (File Eutherini PP D S1) were selected as outgroups, as previous analyses sug- Acemyini EE T Masiphyini – ?P E gest a close affinity between at least some members of the Strongygastrini PPa D Rhinophoridae, Calliphoridae and Tachinidae (Wood, 1987a; Euthelairini – Eb T Kutty et al., 2010; Marinho et al., 2012; Singh & Wells, 2013). Palpostomatini TT D Wherever possible the egg, first-instar larva and adults of both Litophasia PD – sexes were examined for each species, or information on all a stages was retrieved from reliable sources in the literature (File Including Rondaniooestrini. b As Neominthoini. S1). The rhinophorid and most of the tachinid specimens cho- P, Phasiinae; D, Dexinae; T, Tachininae; E, Exoristinae. sen for coding were identified to species by one of us (P.C.), Taxa with at least one representative included in the present study are and the calliphorids were identified by Knut Rognes (Oslo, shown in bold. Norway). These specimens are deposited in P.C.’s collection at the Museum of Zoology, University of Rome ‘Sapienza’. Phylogenetic analysis Specimens of a few rarer species were examined and coded from other collections: CNC, Canadian National Collection of Cladistic analyses using parsimony were conducted with Insects, Agriculture and Agri-Food Canada (Ottawa, Canada); a data matrix of the states of 135 characters belonging to SMNS, Staatliches Museum fur¨ Naturkunde (Stuttgart, Ger- species of 180 tachinid genera, as well as three calliphorid, many); and TAU, Department of Zoology, Tel Aviv University one rhiniid and two rhinophorid genera (Files S1–S3). The (Tel Aviv, Israel) (File S1). The majority of species are from matrix was produced in mesquite version 2.74 (Maddison & the Palaearctic Region, but species from other regions are Maddison, 2010) and analysed in tnt version 1.1 (Goloboff included to provide a global representation of tachinid tribes et al., 2003). The ‘New Technology Search’ (NTS) options (File S1). were used because of the large number of taxa. To find the most We selected 135 morphological characters, distributed parsimonious trees, all analyses were run with the following as follows: egg, nine; first-instar larva, four; and adult, parameters: general RAM of 1000 megabytes and memory set 122 (including 36 in the male postabdomen and 10 in the to hold 600 000 trees; multistate characters were treated as female postabdomen) (File S2). Morphological terminol- unordered and zero-length branches collapsed. The ‘driven ogy essentially follows that proposed by McAlpine (1981) search’ was performed by using 14 replications as the starting (adult external morphology and chaetotaxy), Tschorsnig point for each hit [which is the maximum value as suggested (1985) and Tschorsnig & Richter (1998) (male and female by the program for the most exhaustive search, i.e. when the terminalia) (see also File S2 and explanatory figures initial level is set as 100 from ‘set initial level’ dialog box therein). (= maximum value) or 10 from command line (= maximum

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 Phylogeny of Tachinidae 339 value)]. Each replicate, initially autoconstrained (previous and two homoplasious apomorphies (80:0; 120:1). Monophyly is Wagner), was conducted with: constraint and random based well supported by a Bremer support value of 10 and a resam- sectorial searches, no ratchet, tree-drifting (six iterations) and pling probability of 95. The calliphorid outgroups formed a tree-fusing (10 rounds); finding minimum length trees 30 times, paraphyletic grade, consistent with the findings of Rognes not consensusing trees during search, multiplying trees by (1997), and the two rhinophorids + Rhyncomya (Rhiniidae) fusing after hitting best score and saving one tree per replicate. were reconstructed as sister group to Tachinidae based on three tnt commands from command line were as follows: ‘hold homoplasious apomorphies (51:1; 52:0; 71:1). 600000; xmult = level 10 hits 30 rss css fuse 10 drift 6;’. Analyses were conducted with equal weighting and also with Basal taxa. The Coleoptera-parasitizing clade B implied weights under the default function k/(es + k), where (Gnadochaeta Macquart + Palpostomatini) is reconstructed as k = concavity constant and es = extra steps, with k varying sister group of clade C (i.e. all the remaining tachinids). Mono- continuously between 1 and 10, then for k = 15, 20, 40 (tnt phyly of clade C is weakly supported by two homoplasious command: hold 600000; Piwe = 1, 2, 3, ...n; xmult = level 10 apomorphies (30:0; 43:2). The macquartine Macroprosopa hits 30;) in order to examine and compare tree length, total fit Brauer & Bergenstamm (clade D) is sister group of and variation in tree topology under various weighting schemes clade E ((Ormiini + (Dexiinae + Phasiinae)) + (Tachini- (File S5). Total fit was calculated for most parsimonious trees nae + Exoristinae)) based on three homoplasious apomorphies (MPT) obtained under equal weights for each k-value tested (57:0; 79:1; 111:2). and compared with total fit of MPTs obtained under implied Monophyly of clade B is well supported by two homopla- weighting for corresponding k-values (File S5). sious apomorphies (33:1, 2; 111:1) and one non-homoplasious Bremer support values were calculated in tnt from 10 000 apomorphy (105:1, medial surface of male pregonite mem- trees up to 10 steps longer than the shortest trees obtained from branous). Taxa comprising this clade have been placed in the a ‘traditional search’, using the ‘trees from RAM’ setting. A Tachininae by several authors and here occupy the most basal jackknife resampling was carried out with a traditional search position within the Tachinidae. producing 1000 replicates each of 100 random taxa addition Clade F (Ormiini + (Dexiinae + Phasiinae)), which is sup- subreplicates applying tree bisection-reconnection branch ported by three homoplasious apomorphies (51:0; 65:1; 106:1), swapping and saving 10 trees per replication. Jackknife is divided into two subclades: clade G (the tribe Ormiini, here removal probability was set at the default value of 0.36, represented by Aulacephala Macquart and Therobia Brauer) and resampling percentiles were calculated as frequency and clade H (Dexiinae + Phasiinae). Monophyly of clade G differences [group present/contradicted (GC)]. Character is strongly supported by five homoplasious (13:1; 85:0; 88:2; optimization (using unambiguous changes only) was carried 96:1; 113:1) and three non-homoplasious apomorphies (14:1, out in winclada version 1.00.08 (Nixon, 1999). Ancestral ocelli missing or posterior pair strongly reduced; 42:1, female state parsimony reconstruction of host associations was carried with a very swollen and convex prosternal region; 110:1, out in mesquite version 2.74 (Maddison & Maddison, 2010). basiphallus dorsobasally thickened (Tschorsnig, 1985)).

Results The Dexiinae. Clade H (Dexiinae + Phasiinae) is sup- ported by two homoplasious (109:1; 114:1) and two non- Analysis under equal weights homoplasious (115:1, dorsal connection between basiphallus and distiphallus membranous; 116:1, basiphallus and distiphal- The analysis under equal weights and with all charac- lus joined at a right angle) apomorphies. The dorsal mem- ters unordered produced 87 most-parsimonious trees (tree branous connection between basiphallus and distiphallus has length = 1109 steps, consistency index for matrix, CI = 0.181, long been considered a clear synapomorphy of the subfamily = retention index for matrix, RI 0.731): the strict consensus Dexiinae (see historical review). In the present analysis, this tree is shown in Fig. 1. We selected one of the three shortest character state is interpreted as having undergone a reversal, trees with the highest total fit under the whole range of the being secondarily lost in most Phasiinae. k-values tested (Fig. 2) (i.e. tree number 7 of the nexus file, Within clade H, the monophyly of clade I (i.e. the current File S6) to depict inferred character state changes (File S4), as subfamily Dexiinae excluding the Dufouriini) is supported by discussed in the following. three homoplasious apomorphies (34:0; 53:0; 95:1). Although additional research is needed to corroborate the phylogenetic Tachinid monophyly. The Tachinidae (clade A) are recon- position of the genus Phyllomya Robineau-Desvoidy as sister structed as a monophyletic assemblage based on eight non- taxon of clade K (Stomina Robineau-Desvoidy + Dexiini), it homoplasious apomorphies (1:1, egg dorsal plastron absent; is interesting to point out that the Dexiini (clade L) are nested 3:2, egg shell uncoloured; 7:1, ovolarviparous; 10:1, first-instar within the Voriini sensu Herting (1984) and that the genus mandibles reduced; 12:1, first-instar labrum strongly devel- Stomina takes up a position as sister group of the Dexiini oped; 60:1, subscutellum strongly convex; 133:1, female ovisac based on six homoplasious apomorphies (8:1; 13:1; 30:1; 51:1; very long and generally spiralled when full; 134:1, female 65:0; 85:0). The monophyly of clade L is well supported ovisac enveloped by a well-developed tracheal network) and by four synapomorphic character states: one homoplasious

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 340 P. Cerretti et al.

Outgroups

S Pse Onychogonia p

a Thelymorpha a Baumhaueria r l

udogonia o l a

h n

p Dolic z i l

a l Gonia dochaeta

n a Palesi Gaedia i C Lucilia a Pollenia Pro Rhyncomya D hocolon Phyto Rhinophora Gna Melisoneura Masistylum sopea Ciala Eutrixopsis e ErythroceraOcyt sa Macroprosopa Pachystylum Pal Aulacephala Therobia x es Phyllomya ata 98/6 Stomina Billaea i Myxexoris Erynnia Dexia 52/ Prosena i Elodia /2 Estheria 71/2 n Hebia 92/4 Trixa AllophorocerEuexor 70/2 /2 Zeuxia tops Thelaira a Campylocheta ista Eriothrix Zen Blepharomyia e Sturmia a Platymya illia 65/4 Periscepsia 59/6 Athrycia Nea Phryno Hypovoria lsomyia Hyleorus Masicera Voria Chetoptilia Fro /3 Eumeellantina Dufouria /2 /3 59/2 Pandelleia e Eum /2 91/4 Rondania Cyzenea Microsoma a Bot is 99/8 Eugymnopezaea Winthemiahria Frera /2 Litophasia n RhaphiochSmi dtia 75/2 Imitomyia ster i Phorocer aeta /3 /6 Strongyga P osoma 61/3 t Nem Catharosia orilla Clytiomya Prosethilla h s /2 Ectophasia Ethilla i 95/10 /2 Gymnosoma Medina /3 55/2 Opesia a r 62/3 Phorinia /2 Phasia Bessa 81/2 95/4 Trichopoda s o /2 Chetogena Xysta Phorocera Hermya i

x Exorista Parerigone i

62/3 Weberia n

Staurochaeta E

/2 89/3 Brullaea

Lecanipa 79/2 /3 Clairvill a ia Leiophora Besse

ria Oswaldia 61/3 Cylindromyia e Hemyd Lomachantha /2 Tryphera Lophosiaa Phania Hubneria 93/7 Carcelia /3 Anthomyiopsis ia Graphogaster Paratryphera Heraultia Atylomy Phytom Lydella /4 /3 Ross Drino ypt /2 /2 /3 Hyperaeaimyiop era Chetina /2 Plesina Pse s 92/6 67/2 Minthoudomintho Thecocarcelia Clausicel Periarchiclops /3 96/5 Lesk 90/8 Bithia PseudoperichaetaCatagonia ia Tlephusa Aphria la Demoticus /2 59/2 Brachymera SenometopiaPtesiomyiaPhryxe Pseudopachystylum Gon Nilea /2 96/5 Phebellia 2 Entomophaga Actia ioc

71/4 Ceranthia 64/

Erycia Ceromya era

ria /2 Siphona 59/ Sarrom T

Gymnophryxe Dexios

Microphtha CompsiluraBlondelia Lydina a

Loewia

EutheraLigeria Lypha Petagnia

Synactia yia c Zaira Eloceria

ia N oma

Admontia s e Picconia h

Redtenbache a p a i

nospila o

nice r e

t aria Istocheta i

e r lma

c t

Cerac a n

e

Acemya e arthria

l

l Trigo p i

Cleo e

Metacemyia Tachina Tri

Panzeria o n

P

Germ

Hyalurgus Schineria

Pelatachina e Nemoraea

Linnaemya a N Chrysosomopsis e

Fig. 1. Strict consensus cladogram obtained from 87 most parsimonious trees under equal weights. Branch support: left, maximum parsimony % jackknife (cut 50%); right, Bremer support values.

(71:3) and three non-homoplasious (97:1, ear-shaped medial The Phasiinae and Dufourini. Clade O consists of the Phasi- side of surstylus; 101:1, boundary between hypandrium and inae + Dufouriini. The monophyly of clade O is supported by hypandrial apodeme very distinct; 122:1, acrophallus with a two homoplasious apomorphies (73:3; 90:1). granulate or lamellate structure). The Dufouriini sensu Herting (1984) (Chetoptilia Rondani, Clade M is supported by three homoplasious apomorphies Dufouria Robineau-Desvoidy, Rondania Robineau-Desvoidy, (47:3; 48:0; 79:3,4). The Voria -group (clade N) is well Pandelleia Villeneuve, Microsoma Macquart, Eugymnopeza supported by four homoplasious apomorphies (43:4; 45:3; Townsend, Freraea Robineau-Desvoidy), which are par- 56:0; 73:0) and one non-homoplasious apomorphy (76:1, asitoids of adult beetles, are paraphyletic with respect to crossvein dm-cu exceptionally oblique). the Phasiinae sensu Herting (1984) (clade Q). Clade P

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 Phylogeny of Tachinidae 341

Outgroups

Spallanzania Pseudogonia

Onychogonia a

r Thelymorpha Baumhauer o ia h haeta

a

i

p

l

i

i

l o

l Gonia c D Dolichocol a

u hala

C L Pollen Gaedia Rhyncomya Pales Phyt Rhinophora Gnadoc e Prosopea Melisoneura Ciala ia Eutrixopsis isa Macroprosopa x Frontina Aulacep Pales on Therobia Nealsomyia a Sturmia Phyllomya Stomina i MyxexoristopEumeel Billaea i Dexia Prosen n Eumea Zeuxia AllophorEuexorista la Estheria Trixa a s Thelaira ocer B Zenillia Campylocheta e Cyzenis a Eriothrix Blepharomyia Bothri Periscepsia Phryno L a Athrycia MasistylumOcytata J K Hypovoria Ery Hyleorus Pachystylumthrocera Voria ria Chetoptilia ia Erynnia Dufou Pandelle ia Elodia N Rondan Hebia M Microsoma Masicera G e I Eugymnopeza Platymya Freraea Ptesi a omyia Litophasia Gymnop myia P Pseudoperi hryxe BJ Imito aster n chaeta Strongyg i Catagonia H P Catharosia h Opesia t Phryxe D Phasia Lydella O a F Xysta s Chetina Q Clytiomya s i Drino R S V Ectophasia Nilea r Gymnosoma T i Phebellia A C W Trichopoda

i Hubneria Pareri

o E U gone n Carcelia BI AC Hermya Tryphera Weberi

x AB a X Brullaea a Paratryphera BB AD Z

Atylomyia Clairvillia E

a Y Besser e Senometopi AE ia α Phania Thecocarcelia BH AA Cylind AZ romyia Periarchiclops BF Hemyd Smidtia BE AX AV Lophosiaa Rhaphiochaetama AF AG Anthomy eroso BG BC Grapho iopsis Phoroc Winthemia Heraultiagaster Nemorilla Phytom a BA AW Rossimyiop Prosethilla yptera Ethill AY AI Hyperaea Medina BD Ples s a AH AJ Pseudominthina Phorinia Bess Mintho AK Clau AL Leski Chetogena sicella o Bithia Phorocera a Exorista Aphri AM Demoticus AO Brachymeraa StaurochaetaLecanipa Pse Leiophora AS AN AP Gonioceraudopachystylum Oswaldia AR Entomophaga Cera Siphona

Blondelia AQ AU Actia nthia

Lomachantha Erycia Ceromya Tlephusa a Sarr

AT Dexiosoma T

acheria ia Microph Lydina omyia

Compsilura Euther Lypha Ligeria Loewia a Petagnia

Synact

Zaira Eloceri Nea Neo Admont c Triarthria Redtenb Picconia Germaria Linnaem S

P thalma

a ia c e

era Istocheta n h l plectops h

i e ia i

Ceracia h n a

t

tachina e

Acemya c e r i

Trigonospila a r

Cleonice i

Metacemyia i

a

T a n

ya Panzer Hyalurgus Pela Nemoraea

i n

Chrysosomopsis a e

Myriapoda Insecta: Heterometabola Insecta: Holometabola Chilopoda - Embioptera Coleoptera larvae Lithobiomorpha Orthoptera Ensifera Coleoptera adults Crustacea Orthoptera Caelifera Lepidoptera (folivores/ Isopoda xylophagous) Dermaptera Diptera larvae non-parasitoids Hemiptera Hymenoptera 'Symphyta'

Ambiguous

Fig. 2. One of the 87 most parsimonious trees obtained under equal weights with ancestral parsimony reconstruction of host associations mapped in colour. Clades discussed in the text are labelled with letters.

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 342 P. Cerretti et al.

(Rondania-group), supported by one non-homoplasious Monophyly of the large clade AF is supported by three apomorphy (128:1, female tergite six long and tubular), takes homoplasious apomorphies (48:0; 81:0; 85:0). This assem- up the position as sister group of clade Q, which is, in turn, blage is divided into two large subclades, AG and AV, rep- supported by four homoplasious apomorphies (20:0; 65:0; resenting the remaining Tachininae and entire Exoristinae, 69:0; 116:0). respectively. Litophasia Girschner, Imitomyia Townsend and Strongy- The tachinine clade AG is supported by three gaster Macquart represent rather basal phasiine lineages. Of homoplasious apomorphies (32:2; 65:1; 114:1). The Leskiini these taxa, hosts are unknown for the first two, while the last (Clausicella Rondani, Leskia Robineau-Desvoidy, Bithia has a rather complex host spectrum that includes adult Formi- Robineau-Desvoidy, Aphria Robineau-Desvoidy, Demoticus cidae (S. globula (Meigen); see Herting, 1960), Heteroptera Macquart) are retrieved as a paraphyletic grade of lineages and a plethora of adult Coleoptera (S. triangulifera (Loew); at the base of clade AH. Clade AH, supported by three see Reeves & O’Hara, 2004). homoplasious apomorphies (52:1; 78:2; 114:0), is divided into Four apomorphic character states support monophyly of two subclades (AI and AL). clade R (Phasiinae – Litophasia): two homoplasious (16:1; Clade AI, composed of the tribes Brachymerini and 114:0) and two non-homoplasious (100:1, medial plate of Siphonini, is supported by three homoplasious apomorphies hypandrium elongated; 102:1, pregonite posteriorly connected (33:2; 85:3; 112:0). Monophyly of the tribes Brachymerini to hypandrium). Clade S (clade R – Imitomyia) is supported (clade AJ) and Siphoniini (clade AK) is supported, respec- by three homoplasious (103:0; 106:0; 115:0) and two non- tively, by four homoplasious apomorphies (34:1,2; 43:5; homoplasious (91:1, posterior margin of sternite 5 not notched; 44:1; 65:0) and by five homoplasious (21:2; 41:0; 73:3; 74:1; 93:1, male sternite 6 symmetrical) apomorphies. The mono- 106:1) and two non-homoplasious (104:1, anterior portion of phyly of clade T (i.e. the Phasiinae sensu O’Hara & Wood, pregonite membranous; 135:1, female with two spermathecae) 2004), the members of which are all parasitoids of Heteroptera, apomorphies. is rather well supported by five homoplasious apomorphies Clade AL represents the Tachinini in the broad sense of (7:0; 117:1; 131:1; 133:0; 134:0). Within clade V, the tribe Tschorsnig (1985: 70), i.e. an expanded concept with respect Phasiini sensu Tschorsnig (1985), here represented by Ope- to Herting (1984) and most other authors, plus Pelatachina sia Robineau-Desvoidy, Phasia Latreille and Xysta Meigen, Meade. This clade is supported by five homoplasious apomor- is not recovered as monophyletic. The Gymnosomatini (clade phies (95:1; 111:1; 113:1; 117:0; 119:1). The Tachinini s.l. are W, thus including Trichopoda Berthold), however, are mono- divided into three main subclades: clades AM and AN sup- phyletic and supported by two homoplasious apomorphies (2:1; ported by two homoplasious apomorphies [(24:4; 66:1,2) and 96:1) and one non-homoplasious apomorphy (123:1, sperm (58:0; 79:2,3), respectively]; and clade AO supported by four duct developed into three well-sclerotized ducts). homoplasious apomorphies (43:5; 49:0; 65:0; 80:0). Monophyly of clade X is supported by two homoplasious In clade AM, the highly apomorphic and moth-parasitizing apomorphies (65:1; 96:1). Parerigone Brauer and Hermya genus Sarromyia Pokorny (cf. Cerretti, 2010) is quite Robineau-Desvoidy represent basal lineages to clade Y (Cylin- surprisingly retrieved as sister to the beetle-parasitizing dromyiini + Leucostomatini) whose monophyly is supported tribe Megaprosopini (Dexiosoma Rondani, Microphthalma by one non-homoplasious apomorphy (125:1, phallus smooth Macquart). and tubular). The tribe Cylindromyiini (clade AA) is supported Within clade AN, the genus Loewia Egger clusters with the by two homoplasious (95:1; 131:0) and two non-homoplasious Polideini sensu O’Hara (2002) (clade AP). The monophyly (130:1, female tergite 7 and sternite 7 fused; 132:1, female of clade AP is supported by two homoplasious apomorphies postgenital plate stylet-like) apomorphies. Monophyly of the (43:4; 69:2). This clade is, in turn, sister to clade AQ, which Leucostomatini (clade Z) is supported by two homoplasious is composed of genera currently included in Herting’s (1984) (16:0; 18:2) and one non-homoplasious (127:1) apomorphies. tribes Neaerini (Neaera Robineau-Desvoidy, Neoplectops Malloch), Ernestiini in part (Eloceria Robineau-Desvoidy, The tachinine grade. Clade AB (all remaining Tachinidae, Synactia Villeneuve) and Bigonichetini (= Triarthriini) i.e. Tachininae + Exoristinae) is weakly supported by three (Triarthria Stephens). The first two of these tribes are homoplasious apomorphies (27:1; 35:3; 58:0). Clade AC, sup- reconstructed as polyphyletic. ported by one homoplasious apomorphy (73:3), reconstructs Clade AO is divided into two subclades: clade AR, sup- the Coleoptera-parasitizing genus Anthomyiopsis Townsend ported by three homoplasious apomorphies (39:1; 52:0; 73:0) as sister group of the polyphagous tribe Graphogastrini and clade AS, supported by one homoplasious apomorphy (Graphogaster Rondani, Heraultia Villeneuve, Phytomyptera (15:1). In clade AR, Germaria Robineau-Desvoidy and Lin- Rondani). Within clade AD, which is supported by three naemya Robineau-Desvoidy (clade AT) cluster together, form- homoplasious apomorphies (81:1; 119:0; 121:1), the current ing the sister group of clade AU (Tachinini s.s.). Monophyly tribe Minthoini (Rossimyiops Mesnil, Hyperaea Robineau- of clade AT is supported by just one homoplasious apomorphy Desvoidy, Plesina Meigen, Pseudomintho Brauer & Bergen- (117:1). Clade AU is supported by three homoplasious apomor- stamm, Mintho Robineau-Desvoidy), although representing phies (24:4; 38:1; 44:1) and one non-homoplasious apomorphy rather basal lineages, forms a grade due to lack of any synapo- (94:1, postabdomen capsule-like). In clade AS, the genera morphic character states. Nemoraea Robineau-Desvoidy (Nemoraeini) and Pelatachina

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 Phylogeny of Tachinidae 343

(Pelatachinini) are nested within a group of Ernestiini sensu description of the more interesting and significant variations is Herting (1984), in part. given in the following (Fig. 3).

A monophyletic Exoristinae. Monophyly of the Exoristi- k = 1 (Fig. 3a). Under these extreme weighting conditions, nae + Eutherini (clade AV) is supported by four homoplasious the genus Litophasia is reconstructed as sister group of all apomorphies (2:1; 3:0; 8:0; 45:3). Within this clade the tribe other tachinids due its lack of a well-developed subscutellum. Acemyini (clade AW) is corroborated as monophyletic with The presence of a strongly convex subscutellum (60:1) repre- strong support of six apomorphic character states: four homo- sents the only non-ambiguous, non-homoplasious apomorphy plasious (58:0; 88:1; 103:2; 112:0) and two non-homoplasious present on the consensus cladogram that supports the remain- (40:1, first flagellomere sharply pointed; 124:1, pronounced ing Tachinidae. In this analysis, Eutherini are reconstructed as a shortening of phallus). The Acemyini are sister to clade AX, rather basal lineage of the Exoristinae, while the Dexiinae take which represents all the remaining Exoristinae. up a position as sister to clade Phasiinae (sensu Herting, 1984; Clade AX (Blondeliini, Eryciini, Ethillini, Exoristini, Goni- thus including Imitomyiini and Strongygastrini) + Tachininae ini, Winthemiini, Eutherini) is supported by two homoplasious (paraphyletic). The Ormiini take up a position of sister group apomorphies (52:1; 121:0). to the Phasiinae. Phylogenetic reconstruction of the lineages of subfamily Exoristinae is ambiguous in our analysis. The Eutherini (clade BA), consisting of two known genera (both included 2 ≤ k ≤ 5 (Fig. 3b). Here, the Phasiinae (sensu O’Hara & in the present analysis), have been placed in either the Wood, 2004, thus excluding Imitomyia and Strongygaster) Phasiinae or the Dexiinae, as discussed earlier. Here, the tribe take up a sister-group position to the remaining Tachinidae occupies a nested position within the Exoristinae. Monophyly (cf. Richter, 1991). The subfamily Exoristinae is broken of the Eutherini is strongly supported by nine homoplasious up into a paraphyletic grade of lineages from which the apomorphies (19:0; 51:0; 78:1; 103:1; 106:1; 111:1; 114:1; monophyletic Acemyini and clade Dexiinae + Tachininae 117:0; 119:1) and one non-homoplasious apomorphy (5:1, egg arose. Under these k-values of the weighting function, with a posterodorsal ‘window’). oviparous groups [i.e. Phasiinae, Winthemiini, Ethillini (in Monophyly of each of the tribes Winthemiini, Ethillini, part), Exoristini and the blondeliine genus Medina]are Eryciini and Blondeliini traditionally ascribed to the Exoristi- reconstructed as a basal paraphyletic grade of taxa from nae is not supported in any of the 87 MPTs, suggesting that which a monophyletic ovolarviparous group arose. More these taxonomic groups should be studied in more detail to specifically, under these k-values, the laying of fully embry- delineate monophyletic groups and revise their classification onated eggs (7:1), together with the very long ovisac (133:1) accordingly. In the tree chosen for discussion (Fig. 2), the and the presence of a network of tracheae enveloping the Blondeliini, Eryciini and Ethillini are each reconstructed as ovisac (134:1), seems to have characterized the common polyphyletic. There is some support for monophyly of Exoris- ancestor of all the ovolarviparous tachinids, thus rejecting tini (clade BD), Winthemiini (+ Phorocerosoma Townsend) both the reconstruction we obtained under equal weights and (clade BH) and Goniini (clade BJ). previous hypotheses that ovolarvipary evolved several times A sister-group relationship is supported between independently from oviparous ancestors (cf. Stireman et al., Exoristini (clade BD) and clade BE (Medina Robineau- 2006; Tachi & Shima, 2010). The membranous, translucent Desvoidy + (Winthemiini + Ethillini (in part))). The tribe egg shell (3:2) is the only non-ambiguous, non-homoplasious Exoristini is supported by three homoplasious apomorphies apomorphy supporting clade Dexiinae + Tachininae. The (71:3; 72:1; 96:1). Clade BF, supported by two homoplasious Eutherini are reconstructed as nested within a paraphyletic apomorphies (50:1; 121:1), reconstructs clade BG (Ethilla Exoristinae (3 ≤ k ≤ 5) or as sister group of clade Tachin- Robineau-Desvoidy + Prosethilla Herting) as sister group of inae + Dexiinae (k = 2). The tribe Ormiini and the genus clade BH (Winthemiini and the ethilline Phorocerosoma). Strongygaster represent rather basal lineages of clade (Imito- The Goniini (clade BJ) are reconstructed as monophyletic myia + (Litophasia + (Dexiinae))), which is in turn supported with support from three non-homoplasious apomorphies by two non-ambiguous, non-homoplasious apomorphies (87:4, (3:1, egg shell brown or dark brown; 6:1, egg microtype; abdominal tergite 5 more than 1.5 times as long as tergite 4; 9:1, egg hatching stimulated by proteolytic enzymes of host 115:1, membranous dorsal connection between basiphallus and mesenteron) and one homoplasious (8:1) apomorphy. distiphallus).

Analysis under implied weights k = 6 (Fig. 3c). Under these k-values of the weighting function, the oviparous groups noted earlier are still recon- Using implied weights analyses resulted in different con- structed as a basal paraphyletic grade of taxa from which sensus topologies depending on the k-values of the weighting ovolarviparous groups arose, with the subfamily Phasiinae rep- function (Fig. 3). As a consequence, it is not possible to com- resenting the most basal tachinid lineage. The most notable ment in detail upon all the various relationships observed in the difference from the scenario discussed earlier is that the tribe resulting trees (especially for distal nodes). Instead, a concise Ormiini takes up the position of sister group of Imitomyia +

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 344 P. Cerretti et al.

(a) (b) (c) Outgroups Outgroups Outgroups oviparous oviparous PHASIINAE s.s. Litophasia PHASIINAE s.s. oviparous oviparous EXORISTINAE EXORISTINAE EXORISTINAE + Acemyini ovolarviparous ovolarviparous EXORISTINAE Eutherini EXORISTINAE + Acemyini + Eutherini Acemyini DEXIINAE (including Dufouriini) 40:1 TACHININAE TACHININAE ovolarviparous (monophyletic ovolarviparous 124:1 116:1 when k = 2) 1:1 TACHININAE 1:1 1:1 10:1 2:1 10:1 Imitomyia 12:1 60:1 10:1 57:0 12:1 Ormiini 12:1 Strongygaster 60:1 14:1 60:1 7:1 7:1 42:1 3:2 133:1 Strongygaster 80:0 133:1 Ormiini 134:1 110:1 134 :1 Eutherini Ormiini 3:2 3:2 Imitomyia 99:1 Litophasia Imitomyia 124:1 Litophasia

Strongygaster 100:1 87:4 115:1 DEXIINAE 102:1 DEXIINAE 116:1 91:1 116:1 (including 93:1 PHASIINAE (including Dufouriini) Dufouriini)

k = 1 k = 2 – 5 k = 6

(d) (e) (f) Outgroups Outgroups Outgroups

Myiophasiini + Myiophasiini + Myiophasiini + Palpostomatini Palpostomatini Palpostomatini 105:1 105:1 Macroprosopa 93:1 PHASIINAE 91:1 PHASIINAE 100:1 100:1 102:1 PHASIINAE 102:1 Strongygaster Strongygaster 91:1 100:1 102:1 Strongygaster 105:1 Imitomyia 115:1 Imitomyia 14:1 116:1 Imitomyia Litophasia 115:1 42:1 Ormiini 116:1 110:1 Dufouriini Litophasia Eutherini Dufouriini Litophasia DEXIINAE (in part) DEXIINAE (in part) 1:1 3:2 DEXIINAE 10:1 116:1 Eutherini 12:1 (including Dufouriini) Eutherini 60:1 80:1 Ormiini Ormiini TACHININAE TACHININAE TACHININAE

Acemyini Acemyini Acemyini

EXORISTINAE EXORISTINAE EXORISTINAE

k = 7–8 k = 9–15 k ≥ 20

Fig. 3. Simplified representation of the strict consensus cladograms (with non-ambiguous, non-homoplasious apomorphies mapped) obtained under implied weights (default function) and concavity factor k varying from 1 to 40: (a) k = 1; (b) 2 ≤ k ≤ 5; (c) k = 6; (d) 7 ≤ k ≤ 8; (e) k = 9, 10, 15; k ≥ 20.

Strongygaster and that this clade represents a basal lineage of (including Imitomyia and Strongygaster) and Exoristinae are clade (Eutherini + (Litophasia + (Dexiinae))). reconstructed as monophyletic, while the subfamily Tachininae remains paraphyletic with respect to Exoristinae. The tribes 7 ≤ k ≤ 8 (Fig. 3d). The general topology resulting from Ormiini and Eutherini are recovered as sister groups of Phasi- these analyses is close to that obtained under equal weights, inae and Dexiinae, respectively. such that the basal tachinid lineage is represented by clade Gnadochaeta + Palpostomatini and the remaining Tachinidae k ≥ 9 (Fig. 3e, f). Under these conditions the topology is are divided into two large assemblages: Dexiinae + Phasiinae nearly the same as that obtained under equal weights. Clade (clade F of Fig. 2) and Tachininae + Exoristinae (clade AB of Gnadochaeta + Palpostomatini is retrieved as sister group Fig. 2, but excluding Eutherini). Remarkably, under these con- of all the remaining Tachinidae (see earlier for k = 7and ditions, subfamilies Dexiinae (sensu Herting, 1984), Phasiinae 8) and the Dexiinae and Tachininae are both reconstructed

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 Phylogeny of Tachinidae 345 as paraphyletic with respect to Phasiinae and Exoristinae, hence a reduction in the strength of the weighting function, respectively. The most notable difference is that the tribe which at the limit converges on the unweighted linear function. Eutherini does not cluster within the Exoristinae, taking up the Under implied weighting, a tree that could increase the fit in sister-group position to clade Dexiinae + Phasiinae (clade H characters with few homoplasies by reducing the number of of Fig. 2). For k ≥ 7, which we examined, Ethilla, Prosethilla, steps could also result in an increased number of steps in Paratryphera Brauer & Bergenstamm and Atylomya Brauer characters with many homoplasies, i.e. the total fit may be (Ethillini) form a monophyletic group. increased at the cost of an increased tree length (Goloboff, 1993; Goloboff et al., 2008). The total fit calculated for MPTs obtained under equal weights for each k-value tested Discussion (tnt commands: piwe = 1, 2, ... 40; fit*;) resulted in trees under implied weighting displaying higher fit than the shortest Reconstructing the phylogeny of this large family of para- obtained under equal weights (File S5). Also, by increasing sitoids, a group characterized by extraordinary morphological values of k, the difference in steps between trees under implied diversity and impressive radiation in nearly all terrestrial weighting and the shortest under equal weights diminishes, as ecosystems, remains a difficult challenge. As mentioned in the do the differences in total fit (File S5). introductory sections, the current classification of Tachinidae With a k-value of 1 (Fig. 3a), hence with strongest penalty at the tribal and subfamilial levels is the result of observations to homoplasy, the analysis yielded trees 87 steps longer than on external morphology and male and female terminalia. the shortest trees under equal weights. This extreme degree The phylogenetic value of these traits has never before been of concavity gives certain characters excessive influence on analysed using a rigorous cladistic approach on a broad scale the tree, resulting in clades strongly contradicted by numerous at the world level. This analysis, using an extensive mor- characters. For example, with k = 1 the ‘dexiine-phasiine’ phological data set for a large cohort of taxa, represents the genus Litophasia takes up the position of sister group to first explicit hypothesis of tachinid phylogenetic relationships. all the remaining tachinids due its unique lack of a strongly This first analysis has yielded a number of intriguing results convex subscutellum (60:1). This character state undoubtedly (discussed later) and also provides phylogenetic data that can represents a reversal for Litophasia, which shares a large suite be tested, modified and supplemented in future studies on of states (i.e. external morphology, male and female terminalia) tachinid evolution. with the dufouriines and the phasiines, thus making the entire Despite the widely accepted subdivision of the Tachinidae reconstruction quite unreliable. into four subfamilies (Table 1), clear-cut definitions of these Trees obtained under implied weights with k-values varying groups remain elusive and specialists are aware that some from 2 to 6 (which are still much better in terms of total fit subfamilies are not monophyletic. As a consequence, certain with respect to those obtained under equal weights; see File S5) genera and tribes have been arbitrarily moved from one sub- reconstructed oviparous Exoristinae as a paraphyletic grade of family to another, based simply upon the opinions of special- taxa from which ovolarviparous groups such as the Tachini- ists (however informed they may be) (Table 2). For example, nae and Dexiinae arose (Fig. 3b, c). This hypothesis conflicts in comparing the regional catalogues of Herting (1984) and strongly with previous classification schemes and morpholog- O’Hara & Wood (2004) – whose authors largely agree on basic ideas about tachinid classification – the following dif- ical hypotheses as well as with preliminary molecular data. In ferences are noted (see also historical review and Table 2): fact, the Exoristinae, which are composed of both oviparous the Eutheriini and Imitomyini were treated in the Phasiinae by and ovolarviparous lineages, have been consistently recovered the former and in the Dexiinae by the latter (as proposed by as monophyletic (see Tschorsnig, 1985; Richter, 1991; Stire- Shima, 1989); the Acemyini were classified in the Exoristinae man, 2002; Tachi & Shima, 2010). Here, characters associated by the former and moved to the Tachininae by the latter; the with egg type and incubation, which characterize large groups Strongygastrini were treated in the Phasiinae by the former and (some tribes and subfamilies), are drawing together otherwise in the Tachininae by the latter. unrelated groups and dominating clade structure due to the In addition to inconsistencies in the placement of taxa at concave weighting scheme. the tribal and subfamilial levels among authors, the placement For k > 6, the deeper nodes begin to stabilize around a of a key species or small genus into the wrong (though widely general pattern (Fig. 3d–f) with the same three basal clades accepted) tribe or subfamily is sometimes overlooked. For [(Gnadochaeta + Palpostomatini), (Dexiinae + Phasiinae) and instance, the genus Litophasia, which is consistently placed in (Tachininae + Exoristinae)] as obtained under equal weights the phasiine tribe Catharosiini, is characterized by a very dif- (Figs 1, 2). The notable differences with respect to the ferent, dexiine type of male terminalia (cf. Tschorsnig, 1985). analysis under equal weights are that: (i) the tribe Eutherini is recovered as sister group of (Dexiinae + Phasiinae) (for k ≥ 9) (Fig. 3e, f) or, within this assemblage, as sister to Equal vs implied weights clade (Dexiinae + Litophasia) (for k = 7 and 8) (Fig. 3d); (ii) for k-values varying from 7 to 15, the macquartine genus Increasing k-values of the default weighting function in tnt Macroprosopa takes up a position as sister to Anthomyiopsis resulted in a gradual loss of concavity of the function, and within the Tachininae grade.

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 346 P. Cerretti et al.

Analyses conducted giving the strongest penalty to homo- and understand the development of parasitoid lifestyles within plasy (i.e. 1 ≤ k ≤ 6) appear to be providing excess confidence the oestroid lineage. in very unlikely trees by too severely overweighting charac- ters that change little and underweighting those that are more labile. In the following discussion we focus mainly on the Phasiines from dexiines? results obtained from the analyses under equal weights and under implied weighting with k ≥ 7, referring occasionally to Our analysis generally supports the subfamily groupings certain topologies obtained under implied weighting and k ≤ 6 Dexiinae + Phasiinae and Tachininae + Exoristinae (Figs 1, 2) that require specific comments. (as first proposed by Shima, 1989; see earlier), with only the Exoristinae and the Phasiinae reconstructed as monophyletic lineages under a wide range of weighting schemes. This Tachinid monophyly and sister group contrasts with the views of Mesnil (1966) and Herting (1966) in which the Tachinidae were grouped into Tachininae + Dexiinae The current analysis does not allow us to fully test the and Exoristinae + Phasiinae based on their oviposition and monophyly of Tachinidae or determine their sister group, egg types. Interestingly, the Dexiinae, which were previously given the inclusion of a relatively small number of outgroup considered a well-established monophyletic assemblage, are taxa. However, the monophyly of the Tachinidae is here reconstructed as paraphyletic with respect to the Phasiinae, corroborated by ten synapomorphic character states, and under both equal and implied weights (k ≥ 9) (Fig. 3e, supported by a Bremer support value of 10 and a jackknife f). Following the traditional definition of the Dexiinae resampling probability of 95 (Fig. 1; File S4). Surprisingly, (Tschorsnig, 1985; Wood, 1987b; Shima, 1989; Tschorsnig & some recent phylogenetic reconstructions of calyptrate Diptera, Richter, 1998), all members of the subfamily share a strap-like employing mitochondrial and nuclear genes but including pregonite (103:1) and have the basiphallus and distiphallus only a few tachinid species, have not strongly supported the joined at a right angle (116:1) through a membranous monophyly of Tachinidae (Kutty et al., 2010; Singh & Wells, connection (115:1). In particular, states 115:1 and 116:1, 2013). characterizing the dexiine-type phallus, are here reconstructed Studies aimed at elucidating phylogenetic relationships as autapomorphic support for the clade Dexiinae + Phasiinae. among all Diptera (McAlpine, 1989; Wiegmann et al., 2011; These states are interpreted here for the first time as Caravas & Friedrich, 2013; Lambkin et al., 2013) or within having undergone a reversal, being secondarily lost in the oestroid Calyptratae in particular (Pape, 1992; Rognes, the Phasiinae. The strongly autapomorphic phallus of most 1997; Kutty et al., 2010; Marinho et al., 2012; Singh & Phasiinae (Tschorsnig, 1985), often characterized by the lack Wells, 2013) have yielded contrasting results in reconstructing of a clear border between basiphallus and distiphallus, lends relationships among oestroid families. Thus, the sister group support to this hypothesis. Interestingly, our results support of Tachinidae has been identified as the Rhinophoridae by the affinities between Dufouriini and Phasiinae identified by Wood (1987a) and McAlpine (1989) and the Sarcophagidae Verbeke (1962, 1963) and Tschorsnig (1985), reconstructing by Pape (1992) and Rognes (1997) using morphology- the former as a grade from which the latter arose (Figs 1, 2; based inference, while more recently, molecular data seem File S4). to converge on some member of the Calliphoridae s.l. Although previously suggested by Herting (1984), our [Kutty et al., 2010 (Polleniinae); Wiegmann et al., 2011; analysis provides the first rigorous support for the subfamily Marinho et al., 2012 (Mesembrinellinae); Singh & Wells, affiliation of Litophasia, Imitomyia and Strongygaster within 2013 (Polleniinae)] as possible sister to the Tachinidae. In the Phasiinae. But, as hypothesized by Tschorsnig (1985), our analyses the Calliphoridae are consistently reconstructed Litophasia takes up a position of sister group of the Dexiinae as paraphyletic (see also Rognes, 1997) and, as already under a wide range of weighting schemes (Fig. 3a–d), in hypothesized by Pape & Arnaud (2001), the rhiniid genus particular, when the Dexiinae are recovered as monophyletic. Rhyncomya is sister group to the Rhinophoridae (Figs 1, 2; Interestingly, under both equal weighting and for the lowest File S4) based on three homoplasious apomorphies (48:0; k-values of the weighting function (Fig. 3a, b), the Eutherini 62:1; 81:1) and one non-homoplasious apomorphy (61:1, lower do not cluster with either the Dexiinae or the Phasiinae. The calypter tongue-shaped). Clade Rhyncomya + Rhinophoridae tribe has long been considered a member of the Phasiinae, is retrieved as sister group to Tachinidae under the entire in part because Euthera species (and later confirmed for range of weighting schemes, giving more credence to the Redtenbacheria insignis Egger) are parasitoids of true bugs hypothesis of Wood (1987a), although not entirely rejecting (Acanthosomatidae, Pentatomidae) (cf. Herting, 1966; Shima, that of Rognes (1997), who suggested the rhiinids as sister to 1999). All Phasiinae, with the exception of Strongygaster,are clade Sarcophagidae + Tachinidae. Interestingly, since Rognes parasitoids of true bugs and all other Tachinidae are parasitoids (1997), the sister-group relationship between Sarcophagidae of other arthropods. But unlike other Phasiinae, Euthera and Tachinidae has not been corroborated. Additional studies Loew and Redtenbacheria Schiner are ovolarviparous (7:1) using both morphological and molecular data and broader (Shima, 1999). Herting (1966) and Mesnil (1966) were both taxon sampling of both tachinids and oestroid outgroups will be convinced that the egg of Euthera is of the planoconvex type needed to definitively establish the sister group of Tachinidae (2:1) (File S2, Figures S1, S2) and placement in the Phasiinae

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 Phylogeny of Tachinidae 347 is warranted despite its ovolarviparous habit. Tschorsnig (1985) (1966) discussed a multitude of morphological and ecological was uncertain about the affinities of the Eutherini, recognizing characters of both adults and first instars of all major tachinid certain similarities with the Dexiinae in the male terminalia, tribes and subtribes, and provided a ‘non-rooted’ graphical but ultimately retaining the tribe in the Phasiinae. Shima representation of the relationships among taxa. Mesnil (1966: (1989) placed Euthera at the base of the Dexiinae, and Shima 881) depicted Myiophasiini and Palpostomatini in a rather (1999, 2006), O’Hara & Wood (2004) and Cerretti (2010) ‘central’, although ambiguous, position from which all other continued the placement of the Eutherini in this subfamily. major groups seem to arise. The long and slender distiphallic We have observed that the long and platiform pregonite in the structure shared by members of clade B recalls that of most Eutherini recalls the pregonite of the Dexiinae, as does the dexiines; but the shape of the pregonite and postgonite, as position and orientation of the epiphallus in this tribe (File well as the shape and position of the epiphallus, recall those of S2, Figure S37). In fact, the only marked difference in the many Tachininae and Exoristinae. Several highly homoplasious male terminalia of the Eutherini with respect to the Dexiinae character states of the adult chaetotaxy seem to be involved in is the sclerotized dorsal connection between basiphallus and placing clade B as sister to all remaining tachinids (File S4). distiphallus, as noted by O’Hara & Wood (2004). The unusual Macroprosopa is particularly mobile across phylogenetic combination of a planoconvex egg shape (2:1) and possession analyses, taking up positions at the base of clade C, or, under of a large and tracheated ovisac in which eggs are embryonated implied weighting, nested within the Tachininae. It is worth (7:1; 133:1; 134:1) appears to play a role in reconstructing noting that the distiphallic structure in Macroprosopa is very the Eutherini at the base of Exoristinae (Fig. 3a), or nested similar to that of Gnadochaeta and of the Palpostomatini, within the Exoristinae (Figs 1, 2, 3b). However, for k-values which is more consistent with the phylogeny obtained under ranging from 6 to 40, the Eutherini are retrieved as sister to equal weights and for k-values ≥ 20 (Fig. 3f). Dexiinae or sister to Dexiinae + Phasiinae (Fig. 3c–f). The The phylogenetic position of the Ormiini remains ambiguous former relationship is only inferred when the Dexiinae are regardless of the weighting scheme, taking up positions as reconstructed as monophyletic (6 ≤ k ≤ 8) (Fig. 3c, d). For the sister to the Phasiinae (Fig. 3a, d), sister to Dexiinae higher k-values (Fig. 3e, f), the Dexiinae fragment into a (Fig. 3b, c) or sister to (Phasiinae + Dexiinae) (Figs 1–3e, paraphyletic grade from which the Phasiinae arose and the f). This ambiguity may be due to the highly autapomorphic Eutherini become the sister group of Dexiinae + Phasiinae. In morphology of this group and we hesitate to make any strong all these reconstructions, characters of the male terminalia play claims as to where this taxon may belong. a crucial role in defining phylogenetic relationships over those All remaining Tachininae are reconstructed as a paraphyletic of the egg and the female genitalia, supporting the hypothesis grade from which the monophyletic Exoristinae arose. Nearly of Shima (1989). all the currently recognized tachinine tribes are reconstructed Adults of the small tribe Strongygastrini resemble members as para- or polyphyletic, as already hypothesized by several of Phasia Latreille, and other characters have led most authors (Tschorsnig, 1985). Only a few tribes (Graphogas- authors to place the genus in the Phasiinae (e.g. Herting, trini, Brachymeriini, Siphonini, Megaprosopini, Tachinini s.s.), 1960; Verbeke, 1962; Mesnil, 1966; Herting, 1984; Tschorsnig, nested within this large grade, received support as representing 1985), where it is placed in our analysis. However, unlike other monophyletic groups (File S4). The Siphonini are well sup- phasiines, species of Strongygaster are not parasitoids of true ported, confirming conclusions of Andersen (1983) and O’Hara bugs (see earlier) and are ovolarviparous (7:1). Shima (1989) (1989), based largely upon the possession of only two (rather placed Strongygaster at the base of the Phasiinae. O’Hara & than the usual three) spermathecae in the female reproductive Wood (2004) and Cerretti (2010) doubted the phasiine affinity system (135:1). The Polideini (sensu O’Hara, 2002) are not of Strongygaster and placed the genus in the Tachininae. retrieved as monophyletic here due to the (weakly supported) inclusion of Loewia in this clade, while the other examined members of the Loewiini (sensu O’Hara & Wood, 2004), Tri- The diverse and complicated Tachininae arthria and Eloceria, are included in a clade with Neoplectops and Neaera of the Neaerini. However, the two synapomorphies It is generally agreed that the Tachininae are the most of the male terminalia used to define the recently restruc- weakly supported tachinid subfamily (cf. Tschorsnig, 1985), tured Polideini (‘sternite 5 V-shaped with microspines on inner being composed of a diversity of morphologically heteroge- margins, and distiphallus with short lateral arms and special- neous taxa. This is corroborated by our analysis, in which ized anterior sclerite’; O’Hara, 2002: 20) were not included in the subfamily is fragmented into a number of para- and the present analysis. Support for the Tachinini s.s. (Peleteria polyphyletic lineages. The Coleoptera-parasitizing clade B Robineau-Desvoidy, Tachina Meigen) is relatively strong, sup- (Gnadochaeta + Palpostomatini) and the highly derived Ormi- porting Mesnil (1966) and most subsequent authors (but not ini (clade G) are widely separated from Tachininae (Figs 2, 3), Guimaraes,˜ 1971) in defining this clade as those Tachininae the subfamily where they are traditionally assigned. with the posterior (here posterolateral) margin of the hind coxa Although not supported by resampling, clade B is recovered setose (83:1). Some other tachinids also have a setose hind as sister to all the remaining tachinids under a wide coxa, but the position of the setae is not the same through- range of weighting schemes. Interestingly, this hypothesis is out the family: in tachinine genera, the setae arise from the similar to that advanced by Mesnil (1966: 881–885). Mesnil posterolateral margin of the hind coxa (83:1), whereas in the

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 348 P. Cerretti et al.

Exoristinae and the genus Sarromyia, they arise from the pos- state is shared with several winthemiines, the second is a terodorsal margin of the hind coxa (82:1) (see File S2). This characteristic feature of the Winthemiini, and the last represents difference in the position of the setae suggests that the two setal a plesiomophic condition with respect to the long telescoping patterns are non-homologous and thus two characters were rec- ovipositor of winthemiines. These features indicate that the ognized in our analysis. composition of the Ethillini has been weakly founded, and its monophyly was questioned by Tschorsnig (1985). Cerretti et al. (2012b) argued that three monophyletic groups can be A monophyletic Exoristinae identified within Ethillini, and these groups are recognized as monophyletic in the current analysis: a Phorocerosoma group Although weakly supported, the clade Exoristi- (here represented only by Phorocerosoma); an Ethilla group nae + Acemyini (AV) is reconstructed as a monophyletic (clade BG), characterized by the laying of unembryonated lineage under a wide range of weighting schemes (Figs 1, 3a, eggs (7:0); and a Paratryphera group (clade BI), which d–f). Notably, the Acemyini, which split from Exoristinae lays embryonated eggs (7:1). The laying of unembryonated under k-values varying from 2 to 6 (Fig. 3b, c), never eggs provides homoplasious autapomorphic support to clade cluster within the Tachininae but rather at their base. Recent BC, wherein the Exoristini are positioned as sister group quantitative phylogenetic analyses of the Exoristinae using of (Medina + ((Winthemiini + Phorocerosoma) + Ethilla molecular data have also found evidence of monophyly for group)). The degree of egg development at oviposition, and the subfamily. Stireman (2002) recovered the Exoristinae as hence the reproductive strategy of these flies, plays a key role monophyletic in most analyses, with the possible exception in the non-monophyly of the Ethillini in our reconstruction, of certain ‘rogue’ taxa, including Ceracia Rondani, Masiphya but the resultant pattern of relationships is still not convincing. Brauer & Bergenstamm, and Phyllophilopsis Townsend. In In fact, the Paratryphera and Ethilla groups share possession their more comprehensive analysis, Tachi & Shima (2010) of an operculate egg shell (4:1), which is a very rare condition were able to resolve a monophyletic Exoristinae as sister to in tachinids (Cerretti et al., 2012b). Under implied weighting Tachininae, similar to the results found here. These results this character state is reconstructed as a unique autapomorphy suggest that the seemingly labile character of a ‘haired’ supporting monophyly of the Paratryphera group + Ethilla prosternum (41:0) used by taxonomists to help define groups group, under a wide range of k-values (i.e. k = 1andk ≥ 7). is remarkably phylogenetically conserved, although it is present in some Tachininae and has been lost from some Blondeliini and a few Goniini. Which came first, the larva or the egg? Phylogenetic relationships within the Exoristinae are largely unresolved and only the Exoristini and Goniini are recon- Within the Tachinidae, ovipary (deposition of unembry- structed as monophyletic in all analyses. Although the Exoristi- onated eggs) is generally interpreted as a plesiomorphic con- nae are the most diverse subfamily of Tachinidae in terms of dition with respect to ovolarvipary (deposition of embryonated numbers of described species, they are the most morpholog- eggs), and it is presumed that the latter evolved several times ically homogeneous subfamily as well. Most of the variation independently (Wood, 1987b; Tschorsnig & Richter, 1998; that does exist appears to be highly homoplasious, with similar Stireman et al., 2006; Tachi & Shima, 2010). In our analysis character states recurring over and over again. The lack of res- under equal and implied weights (k ≥ 7), all treated oviparous olution and rampant homoplasy in this subfamily may reflect groups [all the Phasiinae s.s. (clade T) and clade BC, i.e. the its relatively recent and explosive evolutionary radiation. exoristine taxa Exoristini, Winthemiini, some Ethillini (clade Although employing far fewer taxa than here, Tachi & Shima’s BG and Phorocerosoma) and the blondeliine genus Medina] (2010) molecular phylogenetic analyses generally supported are nested within ovolarviparous assemblages (Figs 1, 2), sug- monophyly of each of the tribes of Exoristinae and indicated gesting the possible need to re-evaluate the evolutionary history sister-group relationships as follows: ((((Goniini + Eryciini) + of ovolarvipary vs ovipary. The counterintuitive interpretation Blondeliini) + Exoristini) + Ethillini). Stireman (2002) reached that ovolarvipary could have characterized an early tachinid similar, if less clear, conclusions, except he was unable to ancestor and that ovipary developed several times indepen- recover a monophyletic Goniini. Neither the Eryciini nor the dently from an ovolarviparous ancestor is a novel hypothesis Blondeliini can be defined on the basis of non-homoplasious generated from this analysis. apomorphies, and our results suggest that they may not be The laying of fully embryonated eggs is the most common monophyletic groups. As noted by Wood (1985), the short first reproductive strategy in the family and our parsimony postsutural, supra-alar seta (49:1) that is so useful for separat- reconstruction shows that its occurrence may be more primitive ing the Blondeliini and Exoristini from most other Exoristinae within the Tachinidae than previously thought. Ovolarvipary might be the primitive state of the character. has several major advantages, the primary being that the egg Another issue emerging from this analysis is the probable encloses a fully formed first instar the moment it is laid. Once polyphyly of the Ethillini as currently defined (Cerretti et al., evolved, ovolarvipary presumably opened new opportunities 2012b). Ethillini are traditionally defined as sharing three for the Tachinidae, for instance, the ability to take advantage character states: strongly convex lower calypter, katepimeron of an indirect oviposition strategy, which is always associated setulose, and ovipositor short and unmodified. The first with ovolarvipary. This means that eggs laid in habitats

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 Phylogeny of Tachinidae 349 frequented by potential hosts hatch almost immediately after Host associations deposition and the larvae actively search for a host (e.g. Dexiini) or wait for a host that might eventually pass by Two major patterns emerge when considering host associa- (e.g. Tachinini). In Goniini, a special type of ovolarvipary tions across our phylogenetic reconstruction: (i) there is general has evolved in which eggs are laid on foliage and must be agreement between host use and monophyly of major lin- swallowed by a phytophagous host, subsequently hatching in eages (see the Results section); and (ii) the host preferences of its midgut through the stimulation of proteolytic enzymes. more basal groups tend to be either hemimetabolous orders or The evolution of indirect oviposition undoubtedly allowed Coleoptera (Fig. 2). More than 60% of tachinids are parasitoids Tachinidae to broaden their host spectrum to species that would of lepidopteran larvae (cf. Cerretti, 2010: 43), but none of the not be encountered by a searching female, because they are, for four basal clades (Fig. 2; B, D, F, AB) or the sister groups of all example, hidden, buried or nocturnal. In groups characterized major, more distal, assemblages, are unambiguously associated by a direct oviposition strategy, the laying of ready-to-hatch with Lepidoptera as the primitive condition. eggs is still advantageous because it minimizes the length of Gnadochaeta and the Palpostomatini (clade B) are both time that eggs are exposed and vulnerable during the delicate Coleoptera parasitoids. Gnadochaeta species attack concealed embryogenesis process. endophytic weevil larvae by means of actively seeking There are also potential costs associated with ovolarvipary, first-instar planidia, as do the Dexiini. Palpostomatines are especially for the adult female parasitoid. Tachinidae that nocturnal, or semi-nocturnal, parasitoids of adult scarabaeoids. + + lay fully embryonated eggs have a long and distensible Clade F (Ormiini (Dexiinae Phasiinae)) is ambiguous, being composed of three assemblages (clades G, I and O) ovisac that serves as an incubator and, as a consequence, with diverse host spectra. Clade G (Ormiini) are nocturnal the time between mating and oviposition is longer than in parasitoids of Orthoptera; female ormiines are able to locate oviparous females. Indirect oviposition is an inherently risky singing male hosts by means of an unusual tympanic strategy because the majority of first instars will perish before organ located between the fore coxae (42:1). Within clade encountering a host, but inevitably this greater risk of mortality I (Dexiinae – Dufourini), the Voriini s.l. (clade M) are is balanced by higher fecundity (O’Hara, 1985; Mellini, 1991; composed of Lepidoptera parasitoids, without exception, while Stireman et al., 2006). The laying of unembryonated eggs clade J has the sawfly parasitoid Phyllomya as sister group comes at a cost, because such eggs are exposed to an of clade K (Stomina + Dexiini). The hosts of Stomina are increased mortality risk while the first instar develops, but unknown, while the Dexiini are, with a few exceptions, this risk might be countered by the benefit of the egg being parasitoids of soil or wood-dwelling beetle larvae. One of the deposited directly onto a potential host. Minimizing the time exceptions is the genus Trixa Meigen, which is unusual among necessary to produce and lay eggs also minimizes the risk of Dexiini in parasitizing caterpillars (Belshaw, 1993; Tschorsnig mortality for females before they can reproduce. Furthermore, & Herting, 1994). The nested position of Trixa in clade L these taxa can afford to produce fewer, larger eggs because suggests a relatively recent shift to larval Lepidoptera in this all that are laid can be certain of reaching the host. We taxon. However, ecologically, Trixa is similar to other dexiines think that various combinations of these factors could have in that its Hepialidae hosts (Belshaw, 1993; Tschorsnig & contributed towards making the switch to ovipary sometimes Herting, 1994) are soil dwellers. Within clade O, the Dufouriini advantageous. form a paraphyletic grade of adult Coleoptera parasitoids from Aside from Goniini, a few other Exoristinae, Dexiini and which the Phasiinae arose (clade Q). The only dufourines that most Tachininae, which lay eggs ready to hatch in habitats do not directly attack adult beetles are those belonging to the frequented by hosts, all other ovolarviparous tachinids lay eggs genus Dufouria, which lay eggs on larvae but emerge from directly on or inject eggs directly into the body of the host, as the adults (Kaufmann, 1933; Mellini, 1964). Unfortunately, in all oviparous tachinids. Given our results, it is conceivable there are no host records for two key basal groups, Litophasia that reversals to oviparity occurred in ancestors characterized and Imitomyia, but the position of Strongygaster, as sister by a direct oviposition strategy. to the Phasiinae s.s. (clade T), is particularly interesting. Interestingly, with k-values varying from 2 to 6, analyses Strongygaster has a diverse host range that encompasses adult under implied weights reconstruct ovipary as a plesiomorphic beetles as well as adult ants (Tschorsnig & Herting, 1994; state with respect to ovolarviparity (Fig. 3b, c), more in line Reeves & O’Hara, 2004; Shima, 2006), whereas the Phasiinae with previous hypotheses (see earlier), but with some important s.s. are exclusively parasitoids of Heteroptera. A shift from differences. Under these implied weights, ovolarviparous adult Coleoptera to Heteroptera may have characterized the groups are retrieved as a monophyletic lineage, rejecting the developmental strategy of the phasiine ancestor. The only other hypothesis that ovolarvipary may have evolved several times tachinids known as Heteroptera parasitoids are the Eutherini. independently from oviparous ancestors. As discussed earlier, Under equal weights, the Eutherini are nested within the this result has been achieved at the cost of reconstructing Exoristinae, thus suggesting that parasitism of Heteroptera Exoristinae as a paraphyletic grade. Morphological and evolved independently in the Eutherini and Phasiinae. As molecular evidence (Tschorsnig, 1985; Stireman, 2002; Tachi discussed earlier, analyses under implied weighting and & Shima, 2010) converges on a monophyletic Exoristinae, k ≥ 6 yield an entirely different and probably more likely making this scenario quite unlikely. phylogenetic scenario, reconstructing the Eutherini as more

© 2014 The Royal Entomological Society, Systematic Entomology, 39, 335–353 350 P. Cerretti et al. closely related to Dexiinae and Phasiinae (Fig. 3c–f), although A recent molecular phylogenetic analysis of the Diptera never as sister to the Phasiinae. Interestingly, however, all suggests that the Tachinidae may have arisen as recently as ancestral state reconstructions of host associations across 30 million years ago (Wiegmann et al., 2011). Under this sce- phylogenetic analyses for k ≥ 6 agree in interpreting parasitism nario, the radiation of tachinids would have experienced a time of Heteroptera as independently evolved in these two tachinid lag with radiations that took place in ditrysian host groups. lineages. Such non-parallel clade diversification, where parasitoids radi- Non-lepidopteran hosts also characterize basal lineages of ated into an existing and independently diversified resource, clade AB (Tachininae + Exoristinae). Species in the genus has recently been documented for other host/parasite associa- Anthomyiopsis parasitize chrysomelid beetle larvae (cf. Hert- tions (Gomez-Zurita´ et al., 2007) and would be necessary to ing, 1960). Similar to Dufouria, Anthomyiopsis species can account for host-use patterns in other tachinid lineages, most complete development in the adult host, although in this genus notably for heterometabolous insect parasitoids (e.g. Ormiini, it seems to be facultative, being triggered by specific environ- Phasiinae, Acemyini). mental conditions (Mellini, 1957, 1964). It is worth noting Caterpillars would seem to be excellent hosts for parasitoids that, among parasitoids of Holometabola, larval-adult para- in general. They are often abundant, exposed (exophytic) and sitoids have been recorded almost exclusively in coleopteran unable to flee from attack. Given these traits, it is expected that hosts (Mellini, 1964), suggesting that these groups may have they would be colonized by parasitoids at the first opportunity. retained some degree of presumably ancestral plasticity. But, if there was a lag in their colonization by tachinids, as is Within clade AB (Tachininae + Exoristinae), parasitoids suggested by the current phylogenetic reconstruction and their of Lepidoptera larvae become dominant at clade α (which estimated age, what was the reason? Also, it is interesting is also the most species-rich clade worldwide) and this to note that many other families of Diptera have adopted the host preference may have characterized the ancestor of this parasitoid lifestyle (e.g. acrocerids, nemestrinids, pipunculids, clade. As a consequence, associations with sawflies [clade conopids, rhinophorids, many phorids and bombyliids, some Pseudopachystylum Mik, Hyalurgus Brauer & Bergenstamm, muscids, sarcophagids and calliphorids), but none have Staurochaeta Brauer & Bergenstamm, Blondelia Robineau- colonized lepidopteran hosts as extensively as have tachinids. Desvoidy (in part), Catagonia Brauer & Bergenstamm, Phe- There are several factors that probably played a role in the bellia Robineau-Desvoidy (in part), Myxexoristops Townsend], radiation of Tachinidae on lepidopteran hosts. The groundplan crickets and grasshoppers (clade AZ, Leiophora Robineau- of the Tachinidae was likely characterized by the formation Desvoidy, Phorocerosoma), beetles [Microphthalma, Cleon- of a respiratory funnel within the host (Clausen, 1940), which ice Robineau-Desvoidy, Chetogena Rondani (in part), Med- is the method by which most tachinids avoid encapsulation ina, Istocheta Rondani, Picconia Robineau-Desvoidy, Zaira during their larval endoparasitic stage. Even with this preadap- Robineau-Desvoidy, Erythocera Robineau-Desvoidy], earwigs tation, the parasitization of larval Lepidoptera must have posed (Triarthria, Ocytata Gistel) and chilopods (Eloceria, Loewia) significant problems. Due to their long evolutionary history (Fig. 2) probably represent secondary shifts from primitively with hymenopteran endoparasitoids, caterpillars may have Lepidoptera-parasitizing ancestors. evolved particularly effective defenses (e.g. encapsulation Interestingly, our cladograms agree in reconstruct- responses; Strand, 2008; Smilanich et al., 2009), which make ing clade α as arising from a grade of taxa for which it difficult for non-coevolved parasitoid taxa to colonize them. hosts are mostly unknown (several Graphogaster, Her- These defenses may have been partly overcome by further aultia, Hyperaea, Plesina) or from taxa characterized by specialized refinements to respiratory funnel formation and/or associations with saprophagous micro-moths (most Phyto- other means to circumvent the hosts’ immune response (e.g. myptera,afewGraphogaster and some minthoines) or with Ichiki & Shima, 2003). Additionally, many caterpillars are Embioptera (Rossimyiops, one Phytomyptera, some unde- known to use secondary chemicals from their host plants for scribed Graphogaster) (Cerretti et al., 2009; and unpublished their own defense (Bowers, 1993; Dyer, 1997) and it is pos- data from the E.S. Ross collection at California Academy of sible that tachinids have had to develop special physiological Sciences, San Francisco, CA, USA). Except for Phytomyptera, adaptations to avoid or tolerate these chemical defenses before most of these taxa are very rarely collected and it is possible they could effectively colonize and radiate upon plant-feeding that they may parasitize rarely reared groups of arthropods. lepidopteran larvae. Similar and independently evolved adapta- The possibility that the major groups of ditrysian parasitoids tions may have allowed other tachinid lineages to radiate onto (i.e. clades M and α) may have arisen independently from other groups of plant-feeding and chemically protected insects ancestors that parasitized Heterometabola or Coleoptera leads such as Heteroptera and certain Coleoptera (e.g. chrysomelids). to the hypothesis that tachinids could have had an early Tertiary In addition to surviving the immune response and chemical origin and that their massive radiation took place later in the defenses of lepidopterans, other adaptations have been neces- Tertiary and Quaternary, after that of angiosperms (which date sary for tachinids to take full advantage of this diverse lineage back to the Jurassic–Cretaceous boundary; Sun et al., 2002) of potential hosts. These include the complex synchroniza- and parallel to that of major ditrysian Lepidoptera lineages tion of development between parasitoid and host, which as (Grimaldi & Engel, 2005). However, Tachinidae are exception- a consequence is one of the determinants of host range among ally rare in the fossil record and there are no unquestionable Tachinidae. Often, hormonal changes within the host that trig- fossils from the Eocene or older (O’Hara, 2013a). ger developmental changes are also used as cues by tachinid

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Bulletin of the British Museum (Natural History) Sciences and Humanities). Thanks to Knut Rognes (Oslo, Entomology Supplement, 21, 221 pp. Norway) and to an anonymous referee for useful comments Crosskey, R.W. (1976) A taxonomic conspectus of the Tachinidae and valuable suggestions on an early draft of this manuscript. (Diptera) of the Oriental region. Bulletin of the British Museum P.C. is grateful to H.-P. Tschorsnig, Franco Mason (Verona, (Natural History) Entomology Supplement, 26, 357 pp. Italy) and Daniel Whitmore (Natural History Museum, London, Crosskey, R.W. (1980) Family Tachinidae. Catalogue of the Diptera UK) for stimulating discussions on tachinid systematics and of the Afrotropical Region (ed. by R.W. Crosskey), pp. 822–882. British Museum (Natural History), London. ecology. D.J.I. was supported by a PhD Fellowship from the Crosskey, R.W. (1984) Annotated keys to the genera of Tachinidae CARIPARO Foundation. 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