Male Terminalia of Diptera (Insecta): a Review of Evolutionary Trends, Homology and Phylogenetic Implications

Home , Anevrina, Apsilocephalidae, Atelestus (fly), Blephariceridae, Bolbomyia, Culicoidea

Insect Systematics & Evolution 44 (2013) 373–415 brill.com/ise

Male terminalia of Diptera (Insecta): a review of evolutionary trends, homology and phylogenetic implications

Bradley J. Sinclaira,*, Jeffrey M. Cummingb and Scott E. Brooksb a Canadian National Collection of Insects and Canadian Food Inspection Agency, Ottawa Plant Laboratory - Entomology, K.W. Neatby Building, Central Experimental Farm, 960 Carling Avenue, Ottawa, ON, Canada K1A 0C6 b Invertebrate Biodiversity, Agriculture and Agri-Food Canada, K.W. Neatby Building, Central Experimental Farm, 960 Carling Avenue, Ottawa, ON, Canada K1A 0C6 *Corresponding author, e-mail: [email protected] Published 25 October 2013

Abstract The male terminalia character system in Diptera is reviewed. The phylogenetic implications of apomorphic changes are traced on published cladograms. New synapomorphies include: anteroventral parameral apodeme for the Tipulomorpha; parameral sheath encompassing desclerotized aedeagus for Neodiptera (exclusive of Axymyiidae); endoaedeagus for Xylophagomorpha + Tabanomorpha. Apystomyiidae are classified as the sister group to the Eremoneura based on four synapomorphies (lateral ejaculatory processes absent, subepandrial sclerite extending from base of hypoproct to phallus, bacilliform sclerites extending to tips of the epandrium and surstyli functionally developed, but not articulated) and lack of eremoneuran synapomorphies (i.e., loss of gonostyli, presence of postgonites and phallic plate). The Diptera sperm pump with a functional ejaculatory apodeme is a possible autapomorphy of Diptera, exclusive of Nymphomyiidae and Deuterophlebiidae. Internal details of the male terminalia of Sylvicola and Mycetobia (Anisopodidae), Hilarimorpha (Hilarimorphidae) and Apystomyia (Apystomyiidae) are newly illustrated and homologies of the aedeagus, paramere and sperm pump of the Tipuloidea are clarified.

Keywords Diotera; male terminalia; male genitalia; phylogeny; Tipuloidea; Neodiptera; Anisopodidae; Hilari morphidae; Apystomyiidae

Introduction It has been nearly 20 years since the first two authors published the final two parts of the “genitalia trilogy” — an outline of the homologies and phylogenetic implications of male genitalia in Diptera (Wood 1991; Sinclair et al. 1994; Cumming et al. 1995). This special issue of Insect Systematics & Evolution, devoted to the phylogenetic significance of insect male genitalia, has provided us with the incentive to review various aspects of male terminalia in Diptera (defined as the primary genital segment or

© Koninklijke Brill NV, Leiden, 2013 DOI 10.1163/1876312X-04401001 Downloaded from Brill.com09/30/2021 12:01:19PM via free access 374 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 segment 9, proctiger, and any modified adjacent anterior sclerites), correct some mis- interpretations and analyze new data. There have been numerous studies on Diptera phylogeny since the early 1990s, due primarily to the rapid rise and popularity of molecular-based analyses, many of which have proposed conflicting phylogenies particularly when compared with morphological data sets. In addition, several studies on the musculature of male terminalia in lower Brachycera and nematocerous Diptera have been published during this period, which are potentially useful in evaluating proposed genitalic homologies. Diptera male terminalia demonstrate the most extreme diversity and greatest variability in structure compared to any other part of the adult dipteran body. It is this extreme morphological diversity that has resulted in difficulties in recognizing homologous structures, particularly at the genus or family level and above. This is especially significant when comparing nematocerous Diptera to cyclorrhaphans. In addition, male terminalia are a key morphological source of characters used to distinguish species in the vast majority of Diptera families and there are few modern taxonomic studies that do not include illustrations of male terminalia to aid in species diagnoses. Comparative studies of Diptera male terminalia and internal anatomy have a long history, dating from the work of Dufour (1851). It is not our intent to provide a detailed history of the many studies and their contributions, as such reviews can be found in Griffiths (1972), Hennig (1973), Wood (1991), Cumming et al. (1995), Shatalkin (1995, 2012) and Zatwarnicki (1996). The following account provides a brief overview and introduction to the major historical studies. Cole (1927) provided a general survey of the diversity of Diptera male terminalia, but his illustrations were small and rather unsatisfactory, as expressed by Hennig (1973). In his textbook on insect morphology, Snodgrass (1935) illustrated males of two species of Diptera in the Tipulidae and Calliphoridae, including musculature, but unfortunately did not recognize the rotation of pregenital sclerites in the calliphorid species. Crampton (1936, 1942, 1944) provided one of the first comprehensive studies of the entire external morphology of the Diptera. He illustrated and described the male terminalia across the order focusing on rotation. External sclerites, pregenital segments and spiracles were illustrated, although he did not show internal connections and structures. In his earlier work, Crampton (1936) detailed the homology of the male terminalia of Cyclorrhapha, recognizing hypopygial circumversion, a term first proposed by Feuerborn (1922) for the characteristic 360° dextral rotation in these flies. He also considered that the surstylus was derived from tergite 9, thereby establishing a tergal or epandrial hypothesis for the origin of these claspers. This interpretation of the surstylus was followed by van Emden & Hennig (1956). Steyskal (1957) introduced the terms protandrium (pregenital) and andrium (terminalia) for descriptions of the postabdomens of acalyptrate Diptera. He presented diagrams of the protandria (ventral view of the pregenital sclerites) illustrating their asymmetry and the position of the spiracles. Subsequently McAlpine (1985) referred to these diagrams of the protandria as protandrograms. Griffiths (1972) presented his periandrial hypothesis on the homology of male cyclorrhaphan Diptera terminalia (see Cumming et al. 1995 for discussion). The main

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 375 dorsal genital sclerite was considered to be a replacement sclerite termed the perian- drium. It and the dorsal clasping structures were considered to be derived from sternite 9. Griffiths (1972, 1981, 1990b) based his homology theory on the a priori assumption of functional continuity in clasping function throughout the Diptera, such that the ventral claspers on sternite 9 (gonostyli) in the lower Diptera became the dorsal clasping structures in Eremoneura. In his initial studies Hennig (1936, 1958) was apparently undecided on which homology theory to adopt, and was still somewhat undecided in Hennig (1973) (Ulrich pers. comm.). It was not until somewhat later (Hennig 1976) that he clearly decided on a tergal, or “epandrial hypothesis”, and provided details and reasons for his conclusions. In this study, Hennig detailed the male genitalia of the family Lonchopteridae and outlined the evolution of the male terminalia in Brachycera, including the musculature. Ulrich (1972) studied the musculature in two species of Empididae and proposed the fusion hypothesis (with the epandrium fused to the gonocoxites) in his assessment of male genitalic homologies in the Empidoidea. This homology concept, which considered the dorsal claspers to be sternal, was in agreement with the early ideas of Hennig (1936). McAlpine (1981) closely followed the epandrial hypothesis of Hennig (1976), but refined some of the homologies, in particular interpreting the postgonites of Cyclorrhapha as homologous with parameres in the lower Diptera. He also accepted Hennig’s (1976) interpretation that dorsal clasping lobes (surstyli) were derived from a division of the tenth tergite. Griffiths (1981, 1990a) was highly critical of the morphological interpretations and phylogeny in the Manual of Nearctic Diptera (McAlpine 1981, 1989; Wood & Borkent 1989; Woodley 1989). In particular Griffiths (1981) cited the ontogenetic studies of Dobzhansky (1930), Black (1966), and Laugé (1968), which indicate the dorsal clasping lobes are derived from segment 9, as evidence invalidating all of McAlpine’s (1981) epandrial homology interpretations. Wood (1991), Sinclair et al. (1994) and Cumming et al. (1995) presented a detailed survey of male genitalic homologies and traced their phylogenetic implications on cladograms across the order. The unique use of colour combined with the artistry of Ralph Idema (see Cumming et al. 2011) helped to lay out the intricate details among families. Cumming et al. (1995) reviewed the two main competing genitalic homology theories (epandrial versus periandrial) and presented an alternative interpretation of male genitalic homologies (revised epandrial hypothesis), which addressed the weak- nesses of the epandrial hypothesis of McAlpine (1981) that Griffiths (1981) had criticized. For example, the outer wall of the surstylus was considered by Cumming et al. (1995) to be derived from tergite 9 and the inner wall from a sclerotization of the intersegmental membrane, rather than segment 10. The musculature of the surstylus as outlined by Ovtshinnikova (1994b) also supported the epandrial origin of these claspers. Zatwarnicki (1996) presented a detailed history of the various homology theories and a new male genitalic homology concept termed the hinge hypothesis. The hinge

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 376 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 hypothesis considered the dorsal clasping structures to be derived from sternite 9 while being attached externally to tergite 9 (the epandrium). The subepandrial sclerite that connects to the inner wall of the clasper structures was considered by Zatwarnicki (1996) to be derived from a fusion of the gonocoxites and renamed the medandrium. The hinge hypothesis has been referred to by Yeates and Wiegmann (1999) as counter- intuitive and has gained little support. The ideas of Cumming et al. (1995) were later modified and updated by Cumming & Sinclair (1996), Sinclair (2000) and Sinclair & Cumming (2006), where postgonites in the Empidoidea were recognized as homologous with those in Cyclorrhapha (originally termed gonostyli in Cumming et al. 1995) and gonostyli were considered lost in Eremoneura. This change in homology is also supported by interpretation of the musculature in the Cyclorrhapha, where the muscles of the postgonites are not homologous with the antagonistic pair manipulating the gonostyli of lower Brachycera (Ovtshinnikova 1994b). Asymmetry in Diptera male terminalia in relation to mating position was reviewed by Huber et al. (2007). Nearly all cases of asymmetry occur within the Eremoneura and are present more extensively in only a few families where they have evolved numerous times independently. Change in male-dominated mating positions (i.e., male above female) requires abdominal twisting and flexing, sometimes resulting in asymmetric contact. The asymmetry is not advantageous, but the newly adopted mating position is and asymmetries evolve as adjustments to these positions (e.g., the right side of hybotid fly genitalia is often much enlarged compared to the left). The reduction of the process of the gonocoxal apodemes as seen within the lower Eremoneura, has also probably evolved to compensate for rotation of the terminalia.

Evolution of features of the male terminalia In this paper we follow the revised epandrial hypothesis as presented in the latest sum- mary of genitalic terms and structures (Cumming & Wood 2009) referring the reader to the earlier works cited above when seeking an understanding of the terms used. In discussions of genitalic musculature, the numbering system of Ovtshinnikova (1989) is followed. For the purposes of this study, we follow primarily the Diptera classification and cladograms presented by Wiegmann et al. (2011) for the nematocerous Diptera (Fig. 1), Woodley et al. (2009) and Sinclair and Cumming (2006) for the lower Brachycera and lower Cyclorrhapha (Figs 2 and 3), and McAlpine (1989) as modified by Wiegmann et al. (2011) for the Schizophora (Fig. 4). In the following sections, the male terminalia synapomorphies are listed (see Appendix), discussed and reviewed. A similar set of characters of the male terminalia are also presented in Lambkin et al. (2013). Several new interpretations are presented and potential new synapomorphies are identified. The synapomorphies are traced on the cladograms listed above to aid in their description and phylogenetic interpretation. On the cladograms black hash marks denote uniquely derived states, gray hash marks denote homoplastic states, and white hash marks indicate reversals.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 377

Fig. 1. Relationships of the lower Diptera after Wiegmann et al. (2011), except for Bibionomorpha s.str. which includes Perissommatidae.

Diptera ground plan As outlined in Wood (1991), there are six main elements of the male terminalia of Diptera: epandrium (tergite 9); hypandrium (sternite 9); paired, two-segmented gonopods derived from appendages of segment 9 termed the gonocoxite and gonostylus; paired, unsegmented parameres flanking the aedeagus; the intromittent organ or aedeagus with basal sperm pump; proctiger comprising the epiproct, hypoproct and cerci surrounding the anus. The internal male gonads are configured with paired testes lacking posterior epididymis; paired vasa deferentia that are U-shaped, joined medially and are continuous with a medially joined accessory gland; and paired ejaculatory ducts, which are joined medially, exit the accessory gland posteriorly and extend to the sperm chamber (Sinclair et al. 2007). A key component in the evolution of Diptera male terminalia has been the development of the sperm pump and associated structures. Mecoptera and Siphonaptera are usually considered outgroups of Diptera (Beutel et al. 2011), and the three orders together comprise the Antliophora, named by Hennig (1981) for the shared presence of a sperm pump. Hünefeld & Beutel (2005) studied the sperm pump of these three orders and concluded that a functional and homologous pumping apparatus was not a groundplan character of the Antliophora. Consequently the sperm pump of Diptera should be interpreted as a unique autapomorphy (Fig. 1, character 3). The internal male genital tract is uniquely configured in Mecoptera, Siphonaptera and Diptera with paired U-shaped vasa deferentia that are fused apically (character 1) and are continuous with the medially joined accessory gland (character 2). Sinclair

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 378 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Fig. 2. Relationships of the Brachycera after Woodley et al. (2009), with Apystomyiidae added. et al. (2007) detailed the configuration and diversity of the male gonads in a survey of Diptera indicating subsequent simplifications in several lineages. The accessory gland complex is termed the seminal vesicle in a study of the internal genital system of Bibio marci L. by Spangenberg et al. (2012).

Nematocerous Diptera Deuterophlebiidae and Nymphomyiidae The male terminalia are very simplified in Deuterophlebiidae. The tubular aedeagus is surrounded by an aedeagal sheath (paramere?) and the sperm pump and ejaculatory apodeme are not present (Courtney 1990; pers. obs.). Presumably sperm is discharged by a muscular ejaculatory duct. The sperm pump in Nymphomyiidae is similarly absent or little developed (Courtney 1994). These are two of the few remaining nematocerous families where the internal male genital tract still needs to be studied (see Sinclair et al. 2007; Borkent & Sinclair 2012).

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 379

Fig. 3. Relationships of the lower Eremoneura after Sinclair and Cumming (2006) and Woodley et al. (2009).

The assignment of Deuterophlebiidae and Nymphomyiidae outside the remaining Diptera was an interesting result in the recent molecular analyses of the nematocerous Diptera by Bertone et al. (2008) and the entire order by Wiegmann et al. (2011). These two families are highly adapted morphologically to fast-flowing waters, masking their phylogenetic classification (see discussion in Wood and Borkent 1989). As noted above, the sperm pump is little developed in either family and characterized by the apparent absence of the ejaculatory apodeme. If the Dipteran sperm pump is not homologous with that of the outgroup orders, it is conceivable that a sperm pump with a functional ejaculatory apodeme is a synapomorphy of the remaining Diptera. Lambkin et al. (2013) presented similar results, but proposed Culicomorpha as the sister group to the remaining non-nymphomyiid Diptera (Deuterophlebiidae was not represented in their analysis). Culicomorpha also lacks an ejaculatory apodeme (see below under “Culicomorpha”). The extreme specialization in Nymphomyiidae makes assessment of morphological characters difficult, with numerous features clearly being adaptations to lotic aquatic habits (Schneeberg et al. 2012). A sister group relationship between Nymphomyiidae and the remaining Diptera, initially proposed by Rohdendorf (1964), was not supported in a recent morphological study of the larvae by Schneeberg et al. (2012).

Tipulomorpha (Tipuloidea + Trichoceridae) Recent molecular studies by Bertone et al. (2008) and Wiegmann et al. (2011) support Hennig’s (1973) proposal of the monophyly of Tipuloidea + Trichoceridae. Wood &

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 380 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Fig. 4. Relationships of the major lineages of Schizophora as defined by McAlpine (1989) and modified by Wiegmann et al. (2011). Monophyly and composition of superfamilies between the two studies is not entirely in agreement.

Borkent (1989) provided a detailed critique of the synapomorphies of Tipulomorpha (sensu Hennig 1973) and assigned Trichoceridae to Psychodomorpha. However, Griffiths (1990a) rejected their arguments and listed four synapomorphies for the sister groups Tipuloidea + Trichoceridae, including reduction of the male cerci and development of gonopods from posterolateral zones of proliferation. The validity of these characters was questioned and reviewed by Woodley et al. (2009). These two opposing

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 381 hypotheses on the phylogenetic classification of Trichoceridae were also reviewed and critiqued by Oosterbroek & Theowald (1991). Subsequently Oosterbroek & Courtney (1995) reanalyzed the phylogeny of the nematocerous Diptera and also considered Trichoceridae as the sister group to Tipuloidea. However, in an analysis of the larval head capsule by Neugart et al. (2009), Trichoceridae showed no affinities with Tipuloidea, but were found to share a specialized mandible with psychodomorph groups as previously indicated by Wood & Borkent (1989) and Sinclair (1992). In addition, adult head structures apparently do not support the monophyly of Tipulomorpha (Schneeberg & Beutel 2011). Despite this controversy, we tentatively propose the following new character that supports the monophyly of Tipulomorpha (sensu Hennig 1973): paramere prolonged anteroventrally as a pair of slender apodemes (character 4) (equivalent to ‘cranialer Apophyse der Paramerenbasis’ sensu Neumann 1958, fig. 9a inTrichocera Meigen) (Fig. 1). This interpretation is supported by muscle M31 (muscle ‘M’ in Neumann 1958), which in this group stretches between the ejaculatory apodeme and the anteroventral parameral apodeme (Ovtshinnikova 1989). The anteroventral parameral apodeme is clearly present in Trichoceridae (Neumann 1958; Ovtshinnikova 1989; Wood 1991, fig. 6a, e), Pediciidae (Sinclair 2000, fig. 5; Ribeiro 2008), Cylindrotomidae and Limoniidae (Fig. 7; Ribeiro 2008). It is also possibly present in Tipulidae (see under Tipuloidea). This character requires further analysis, including detailed surveys of other nematocerous families for possible homologous processes. Fitzgerald (2004) identified similar processes within Bibionomorpha s.lat., but the homology of these features with those of Tipulomorpha is questionable.

Tipuloidea (Pediciidae, Cylindromatidae, Limoniidae, Tipulidae) There are no known synapomorphies of the male terminalia that define Tipuloidea, but the ground plan condition has been hypothesized based on the family Pediciidae being the sister group to the remaining tipuloids (Ribeiro 2008; Neugart et al. 2009; Peterson et al. 2010). The ground plan condition includes the following features: gonostylus simple; aedeagus straight and simple; ejaculatory apodeme not wider than aedeagus; interbases present, clearly noticeable and fused medially; lateral parameral processes present and simple; and anteroventral parameral apodeme present (Ribeiro 2008). An interesting development in Tipuloidea is the bifid gonostylus referred to as the ‘clasper of gonostylus’ and ‘lobe of gonostylus’ (sensu Ribeiro 2006). This condition is considered to have evolved within the lineage, exclusive of Pediciidae (Ribeiro 2008). The homology of these lobes has been confirmed with muscles, where the clasper of gonostylus possesses a pair of muscles (M27 and M28) and the lobe of gonostylus bears a single muscle (M33) (Paramonov 2004). The term interbase is used by tipulid workers to refer to the dorsolateral portion of the paramere that connects or extends to the gonocoxal apodeme. Ribeiro (2008) refers to the pair of anterior lobes projecting from the parameral bridge as interbases. Blade- like posteriorly projecting lateral parameral processes are present in Cylindrotomidae

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 382 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Figs 5–8. Tipuloidea. (5) Dolichopeza obscura (Johnson), aedeagus and paramere, lateral view with apex of aedeagus omitted; modified from Wood (1991, fig. 5c). (6) Dolichopeza obscura, aedeagus, paramere and adminiculum, ventral view; modified from Wood (1991, fig. 5e). (7)Rhipidia lecontei (Alexander), aedeagus and paramere, ventral view; modified from Wood (1991, fig. 4d). (8) Epiphragma fasciapennis (Say), aedeagus and paramere, ventral view; modified from Wood (1991, fig. 2e). Abbreviations: adm, adminiculum; aed, aedeagus; ej apod, ejaculatory apodeme; goncx apod, gonocoxal apodeme; pm, paramere. Scale bars = 0.1 mm.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 383 and some Limoniidae, but are absent in Tipulidae (Ribeiro 2008). Similar paired pro- longations are found in Trichoceridae (Wood 1991, fig. 6e) and Blephariceridae (Sinclair 2000, fig. 1). The adminiculum is another term used by tipulid taxonomists referring to a structure which serves as a support and guide for the aedeagus (Frommer 1963). It appears as a partly membranous invagination from the posteroventral margin of the gonocoxite and is moved by gonocoxite-hypandrial muscles, M33 (Paramonov 2004). Homologous muscles are found in Trichoceridae, Bibionomorpha and Brachycera (Ovtshinnikova 1989). In Dolichopeza Curtis, the adminiculum is greatly lengthened and thickened laterally, extending anteriorly to the sperm pump (Fig. 6). Interpretation of the musculature of the aedeagal complex permits re-evaluation and refinement of the homologies depicted in Wood (1991) for tipuloids. Muscle M31 inserts on the ventral margin of the ejaculatory apodeme and originates on the antero- lateral margin of the paramere, and muscle M30 originates on the gonocoxal apodeme and inserts on the posterior end of the ejaculatory apodeme in Bibionidae and Brachycera (Ovtshinnikova 1989, figs 51 and 207). In Trichoceridae, the origin of M30 (muscle ‘L’ of Neumann 1958) appears to have migrated onto the posterolateral corner of the paramere, near the point of junction with the gonocoxal apodeme (Ovtshinnikova 1989, fig. 202; Neumann 1958, fig. 8). Paramonov (2004) identified M31 and M30 in Tipulidae and if correct the paired apodemes surrounding the ejaculatory apodeme in Dolichopeza (Wood 1991, fig. 5c, d, e) are parameres if we follow the above origin of these muscles (Figs 5 and 6). Based on the musculature of Limonia Meigen in Paramonov (2004), the outer aedeagal sheath is parameral in origin and the aedeagus is restricted to the inner tube and ejaculatory apodeme, as illustrated in the related genus Rhipidia Meigen (Fig. 7). In addition, the homology of the aedeagus in Epiphragma Osten Sacken in Wood (1991, fig. 2) should be restricted to the inner tube with the ejaculatory apodeme at its base (Fig. 8). The surrounding triangular components are parameral in origin (see Ribeiro 2008, fig. 168).

Trichoceridae This is the only nematocerous Dipteran family where three muscles associated with the sperm pump have been identified (Ovtshinnikova 1989; Sinclair et al. 1994, p. 412). Muscle M32 originates on the ejaculatory apodeme and is inserted on the lateral ejaculatory processes in lower Brachycera, whereas in Trichoceridae this muscle appears to be inserted on an apodeme of the aedeagus. The homology of this muscle in Trichoceridae needs to be re-evaluated.

Psychodomorpha The monophyly of this infraorder (which includes Psychodidae, Blephariceridae and Tanyderidae) is strongly supported in the molecular analyses of Bertone et al. (2008) and Wiegmann et al. (2011). The phylogeny of the Psychodidae was recently inferred

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 384 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 from nuclear DNA sequences (Curler & Moulton 2012). In this study they reviewed the morphological evidence for the relationships they proposed and included a discussion of male genitalic characters. The cerci appear to be absent in Psychodinae and they refer to the lateral clasping lobes as surstyli (Curler & Moulton 2012). We basically agree with their interpretation, although we consider these articulated lobes in Psychodinae as epandrial in origin. The “lateral lobes” of the Phlebotominae should be termed epandrium, but differ from the Psychodinae in not being articulated toward the apex.

Culicomorpha As discussed in previous reviews, this infraorder remains the most strongly defined lineage in the nematocerous Diptera (Yeates et al. 2007). Among the synapomorphies are a number of male genitalic characters (Fig. 1) including presence of a ventral plate (character 7), a membranous aedeagus (character 8), and lack of an ejaculatory apodeme (character 9) (Sinclair et al. 2007). The homology of the paramere and aedeagus in Culicomorpha was revised by Sinclair et al. (2007). Male genitalic synapomorphies are further discussed by Borkent (2012). The development of a multi-chambered accessory gland complex (character 6) was considered by Sinclair et al. (2007) to be a synapomorphy of Bibionomorpha s.str. + Ptychopteridae + Culicomorpha, and is correlated with the production of a spermatophore. Further study of this character is required, especially histological studies exam- ining and comparing cell types in each chamber to determine their homology. Based on the phylogeny of Wiegmann et al. (2011), a multi-chambered accessory gland appears to have evolved independently three times (Fig. 1, character 6). Culicoidea (Corethrellidae + Chaoboridae + Culicidae) remains a very strongly supported group with the male genital tract reversing to a more simplified condition (Sinclair et al. 2007; Borkent et al. 2008). It was postulated in the latter study that a simple genital tract is correlated with permanent rotation of the terminalia between segments 7 and 8.

Neodiptera (Bibionomorpha s.str. + Axymyiidae + Anisopodidae + Scatopsoidea + Brachycera) The sister group relationship between Brachycera and Bibionomorpha s.lat. (including Axymyiidae, Anisopodidae and Scatopsoidea) was strongly supported in the molecular analyses of Bertone et al. (2008) (exclusive of Axymyiidae) and Wiegmann et al. (2011) (inclusive of Axymyiidae). Hennig (1973) was the first to suggest a sister group relationship between Bibionomorpha s.lat. and Brachycera based on modifications of the pleural sclerites, but these characters remain unsubstantiated. Michelsen (1996) proposed several convincing characters of the pronoto-cervical region supporting the monophyly of this group including Axymyiidae, which he termed the Neodiptera. We have re-evaluated the evolution of the parameral sheath in the lower Diptera and propose herein a new synapomorphy supporting the Neodiptera (Fig. 1), exclusive of Axymyiidae. In this clade, the parameres are fused dorsally and form a sheath,

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 385

Figs 9–12. (9) Sylvicola fuscatus (Fabricius), aedeagus and paramere, lateral view. (10) Mycetobia sp. nr. divergens Walker, terminalia, lateral view. (11) Mycetobia sp. nr. divergens, aedeagus and paramere, lateral view. (12) Mycetobia sp. nr. divergens, aedeagus and paramere, dorsal view. Abbreviations: aed, aedeagus; aed gd, aedeagal guide; cerc, cercus; ej apod, ejaculatory apodeme; ej dt, ejaculatory duct; lat ej proc (?), lateral ejaculatory process (?), pm, paramere. Scale bar = 0.1 mm.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 386 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 encircling the aedeagus (character 5). The derived condition is readily observed in the terminalia of Hesperinus Walker and Xylophagus Meigen (Sinclair 2000, figs 8, 10 and 11; Sinclair et al. 1994, fig. 4). In addition to the dorsal fusion, the paramere (or parameral sheath) encases the aedeagus, such that at most only a short portion of the aedeagus protrudes posteriorly. The encasing results in desclerotization of the aedeagus, which is often reduced to a simple membranous inner tube or endophallus. The parameral sheath is termed the dorsal sclerite in Spangenberg et al. (2012), who proposed it as a synapomorphy of Bibionomorpha + Brachycera. The enclosing and fusing of the parameral sheath around the apex of the aedeagus to form a modified intromittent organ or phallus, was hypothesized by Sinclair et al. (1994) to support a sister group relationship between the brachyceran clades Stratiomyomorpha and Muscomorpha (sensu Woodley 1989). We propose here that the initial stages in the development of the phallus evolved earlier within the nematocerous Diptera. Elsewhere among nematocerous Diptera, the paramere does not encircle the aedeagus and the aedeagus remains well sclerotized and separate (e.g., Axymyiidae, Trichoceridae, Blephariceridae, Ptychopteridae, Tanyderidae, Psychodidae, most Tipulomorpha). Although the parameres are fused dorsally in Axymyia McAtee, this is not viewed as homologous to the parameral sheath because the aedeagus remains independent and sclerotized (see Wood 1991, fig. 9e). Blephariceridae have a similar encasing paramere, but the aedeagus remains distinctly separate and tubular, and is not encircled (see Sinclair 2000, fig. 2). Given the strong morphological and molecular support for Neodiptera (Michelsen 1996; Bertone et al. 2008; Wiegmann et al. 2011), lack of a parameral sheath in Axymyiidae suggests a possible sister group relationship with the rest of the Neodiptera rather than a reversal in this character as implied by the phylogeny of Wiegmann et al. (2011) (Fig. 1, character 5:0). The migration of the origin of muscle M30 from the paramere (as discussed above under Tipuloidea) to the gonocoxal apodeme is known in the Bibionomorpha s.str. and Brachycera (Ovshinnikova 1989, figs 51 and 207). This is another potential synapomorphy of this clade, although additional nematocerous families need to be investigated.

Bibionomorpha s.str. (Pachyneuridae, Bibionidae s.lat., Perissommatidae, Sciaroidea) The male genital tract is highly modified in all families of Bibionomorpha s.str. (unknown for Perissommatidae) and undoubtedly reflects the production of spermatophores (Blaschke-Berthold 1994; Sinclair et al. 2007). The formation and transfer of spermatophores in Bibionomorpha s.str. is characterized by an ejaculatory apodeme that is positioned ventral to the sperm chamber (character 10) so that it can be drawn in and out to push out a spermatophore as it forms (Sinclair et al. 2007). This derived modification of the ejaculatory apodeme (often labelled as the aedeagus in taxonomic papers) is a synapomorphy of all families of the Bibionomorpha str., including Perissommatidae (Fig. 1). In other Diptera the ejaculatory apodeme is positioned anterior to the sperm chamber, acting as a piston to compress the chamber to eject sperm.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 387

In the little known family Perissommatidae, the ejaculatory apodeme is a long slender apodeme closely associated with the expanded medioventral margin of the gonocoxites (see Colless 1969, fig. 1A, ap). The paramere comprises three components, a dorsal sclerite, a pair of lateral sickle-shaped rods, and a ventral thickly sclerotized inverted U-shaped brace (see Colless 1969, fig. 1A, dl, pa, ts). The structure labelled as the aedeagus by Colless (1969, fig. 1A, ae) is actually the medioventral margin of the gonocoxites. Sickle-shaped parameral rods are also present in Bibionidae and the pachyneurid genus Cramptonomyia Alexander (Ovtshinnikova 1989; Wood 1991; Blaschke-Berthold 1994). As stated by Colless (1969) the genitalia of Perissommatidae clearly show derived attributes of the Bibionomorpha s.str. This family was assigned to the Psychodomorpha on the basis of larval head characters (Wood & Borkent 1989), whereas the assignment of Perissommatidae was ambiguous in Bertone et al. (2008), or assigned as sister group to Bibionomorpha s.lat. + Brachycera in Wiegmann et al. (2011). Amorim (1993) assigned the family along with Axymyiidae and Pachyneura Zetterstedt as sister group to the remaining Bibionomorpha s.lat. + Brachycera.

Axymyiidae, Anisopodidae and Scatopsoidea (Scatopsidae + Canthyloscelidae) The male genital tract of Axymyiidae (Borkent & Sinclair 2012), Anisopodidae and Scatopsidae (Sinclair et al. 2007) is not highly modified and lacks a multi-chambered accessory gland (character 6) present in those Bibionomorpha s.str. known to produce spermatophores. The internal configuration and external genitalia with a narrow aedeagus and often a long filamentous endophallus, also suggests that spermatophore production does not occur in these three families. There is a trend in the reduction of the epandrium that may indicate phylogenetic arrangement in these families. The epandrium ranges from split dorsomedially in Apistes Meigen (Scatopsidae) and Axymyia (Wood 1991, fig. 9b) to partial or complete fusion with the gonocoxites and hypandrium in Mycetobia Meigen (Fig. 10) and Sylvicola Harris (Anisopodidae).

Anisopodidae + Scatopsoidea A comparative study of the male terminalia and musculature of Anisopodidae and Scatopsoidea is needed. There are very few studies of the male terminalia of Anisopodidae that detail the internal aedeagal-paramere complex. General details for some taxa can be observed in Abul-Nasr (1950), Dahl (1980) and an unpublished thesis by Fitzgerald (2004). In addition, no studies of genitalic musculature are available for either the Anisopodidae or Scatopsoidea. These two family groups are characterized by a long filamentous endophallus (Fig. 9; Cook 1981, fig. 18; absent inMycetobia ), a parameral sheath with paired apodemes anteriorly (Figs 9 and 11), and paired obliquely projecting oval apodemes associated with the sperm chamber (Fig. 12). All three features are possible synapomorphies for Anisopodidae + Scatopsoidea, which is a sister grouping previously proposed by Wood & Borkent (1989) on the basis of the reduction of the hypostomal bridge in the larval head capsule.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 388 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Fitzgerald (2004) considered the divergent paired apodemes adjacent to the sperm chamber (Fig. 12; Cook 1981, fig. 18) observed in both family groups to be homologous to the lateral ejaculatory processes found in Brachycera (character 11). This possible homology was tentatively rejected by Sinclair et al. (1994) based on the apparent posterior position of the apodemes in Anisopodidae and Scatopsoidea compared with an anterior position in Brachycera. This potential homology requires further study in light of the discovery of similar paired apodemes in Mycetobia (Fig. 12), as it could be important in clarifying the origin of Brachycera. The family Valeseguyidae is currently assigned to the Scatopsoidea (Amorim & Grimaldi 2006). Only a single damaged male specimen of the extant species Valeseguya rieki Colless is known and on the basis of available illustrations (Amorim & Grimaldi 2006, fig. 6b, c), this species appears to share features of both the Scatopsoidea and Anisopodidae (originally assigned to the latter family). The paired “aedeagal apodemes” in Valeseguya Colless are possibly homologous to the dorsal anteriorly projecting processes of the paramere seen in Sylvicola (Fig. 9). Additional male specimens of this poorly known taxon are required to carefully evaluate the homology of the internal male terminalia.

Brachycera The Brachycera (Figs 1 and 2) are a well supported monophyletic and long recognized suborder (Woodley 1989; Woodley et al. 2009) currently defined by at least two genitalic characters: presence of paired lateral ejaculatory processes (character 11), and epandrium and hypandrium separate (character 12) (Sinclair et al. 1994). The lower or orthorrhaphous Brachycera have been considered paraphyletic in relation to the remaining Brachycera since Hennig (1973) (Lambkin et al. 2013). However, recently the concept of a monophyletic Orthorrhapha exclusive of Empidoidea, has been resurrected (Wiegmann et al. 2011; Pape et al. 2011). No morphological characters are currently known that support this lineage. Stratiomyomorpha The monophyly of this infraorder is supported by one character of the male terminalia: sperm pump lying on a long concave parameral sclerite (character 13) (Sinclair et al. 1994). In this condition the sperm pump is cradled in a concave anterior prolongation of the parameral sheath (not aedeagal sclerite). Palmer et al. (2000) summarized the musculature of the male genitalia for the Stratiomyomorpha. Several convincing characters of the larval head capsule also support the monophyly of the Stratiomyomorpha (Sinclair 1992). Pantophthalmidae Sinclair et al. (1994) stated that the lateral ejaculatory processes were lost in the Stratiomyomorpha (along with loss of muscle M32), but Ovtshinnikova (1994a) found that M32 is retained in Pantophthalmidae, presumably inserted on reduced lateral ejaculatory processes.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 389

Xylomyidae + Stratiomyidae The monophyly of this clade is supported by the following characters: lateral ejaculatory processes absent (character 11:0); sperm pump reduced, lying ‘free’ on parameral sclerite (character 14:3) (Sinclair et al. 1994). Associated with the absence of the lateral ejaculatory processes and reduction of the sperm pump, is the loss of muscles M30, M31 and M32 in both families (Ovtshinnikova 1986, 1994a). Small muscles associated with the sperm pump are present in Stratiomyidae, which may be homologous with remnants of muscle M32 (Ovtshinnikova 1994a). The reduction of the muscles associated with the ejaculatory apodeme in this clade is unique among the lower Brachycera.

Xylomyidae The gonocoxites of Xylomyidae each terminate in an outer, partially articulated apical lobe and an inner, fully articulated subapical lobe (Sinclair et al. 1994, fig. 11a). Ovtshinnikova (1986) identified the outer lobes as gonostyli based on the insertion of a single muscle (M27) and considered the inner lobes, which lack musculature, as parameres following McAlpine (1981, fig. 122). However, in a later study Palmer et al. (2000) stated that both pairs of gonostylar muscles (M27 and M28) are absent in Xylomyidae. In contrast, Sinclair et al. (1994) identified the inner fully articulated lobe as the gonostylus rather than the outer lobe. The homology of the inner and outer lobes of the gonocoxites in Xylomyidae needs to be re-evaluated.

Tabanomorpha + Xylophagomorpha The endoaedeagal process is a narrow projection directed posteriorly from the ejaculatory apodeme that extends into the endophallus (Theodor 1976). Sinclair et al. (1994, fig. 3a) originally proposed the presence of this character as a synapomorphy of the Brachycera. Griffiths (1996) criticized this coding, stating that due to its rather rare occurrence elsewhere in the lower Brachycera, it was more parsimonious to view the presence of the endoaedeagal process as a synapomorphy for Tabanomorpha + Xylophagomorpha (character 15). We have accepted this interpretation and now assume that the occasional presence of similar processes in other lower Brachycera (e.g., in some Nemestrinidae (see also Richter & Ovtshinnikova 1996), Bombyliidae and Asilidae) represent non-homologous structures. For instance, Cannings (2002) has indicated that the endoaedeagal process in species of Lasiopogon Loew (Asilidae) appears to be simply the enlarged base of the endoaedeagal tube.

Tabanomorpha The monophyly of the Tabanomorpha (inclusive of Vermileonidae) is supported on the basis of the retractile larval head capsule and features of the female cercus (Zloty et al. 2005) and molecular evidence (Wiegmann et al. 2000). The Tabanomorpha have recently been classified into two superfamilies, Rhagionoidea and Tabanoidea (including Vermileonidae) (Kerr 2010). In contrast, the Vermileonidae were assigned as sister group to the Rhagionidae in Wiegmann et al. (2000) and Wiegmann et al. (2011).

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 390 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Within the Tabanomorpha, support for the monophyly of the Rhagioninae is sug- gested by a laterally compressed endoaedeagal process (Kerr 2010). The clade including Oreoleptidae + (Athericidae + Tabanidae) is supported by several male genitalic features, including the fusion of the hypandrium and gonocoxites (character 16), lateral ejaculatory processes closely associated with anterior apex of tines (character 17:2), presence of aedeagal tines (character 18) and greatly lengthened gonocoxal apodemes (character 19:1) (Zloty et al. 2005). Aedeagal tines are also present in Bolbomyia Loew, but are likely independently derived (Stuckenberg 2001; Zloty et al. 2005). The Oreoleptidae + (Athericidae + Tabanidae) clade is also supported in the analyses of Kerr (2010) and Wiegmann et al. (2011). In addition, Stuckenberg (2001) and Zloty et al. (2005) list the epandrium shape and male cercus position as synapomorphies supporting the sister group relationship of Athericidae and Tabanidae.

Muscomorpha (Nemestrinidae, Acroceridae, Heterodactyla) The Muscomorpha was proposed by Woodley (1989) and supported by three adult non-genitalic characters. The monophyly of the Muscomorpha is also corroborated by several male terminalia characters: accessory gland and vasa deferentia not continuous (character 2:0); epandrium articulated with gonocoxites or hypandrium (character 20); gonostyli articulated obliquely or dorsoventrally (character 21); aedeagus and parameral sheath fused, forming phallus (character 22) (Sinclair et al. 1994). The gonostyli articulate horizontally in the therevoid family Evocoidae (Yeates et al. 2003), which is likely a reversal. The development of a phallus (character 22) has apparently occurred independently within the higher Stratiomyidae and Xylomyidae. See Ulrich (2006) for an opposing view on the structure of the intromittent organ of Muscomorpha.

Nemestrinidae and Acroceridae Gonostyli are lost or fused with the gonocoxites (character 23) in Acroceridae and in at least one nemestrinid, Atriadops javana (Wiedemann) (Sinclair et al. 1994). The oblique direction of gonostylar movement (character 21) was confirmed in Nemestrinidae by Richter & Ovtshinnikova (1996), contrary to the opinion of Griffiths (1996).

Heterodactyla (Bombyliidae, Hilarimorphidae, ‘Asiloidea’, Eremoneura) The monophyly of the Heterodactyla is supported by a single character of the male terminalia: epiproct absent (character 24) (Sinclair et al. 1994). Within the Heterodactyla, the Bombyliidae (exclusive of Mythicomyiinae and Heterotropus Loew) is supported by the reduction of the processes of the gonocoxal apodemes (Sinclair et al. 1994). This interpretation and definition of the Bombyliidae is also supported by recent molecular evidence (Trautwein et al. 2011) and larval characters (Yeates & Irwin 1992; Krivosheina 2012).

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 391

Figs 13–14. Hilarimorpha. (13) Hilarimorpha sidora Webb, hypandrium, phallus and gonostylus, lateral view. (14) Hilarimorpha ditissa Webb, terminalia, ventral view (modified from Webb 1981, fig. 4). Abbreviations: apod, apodeme; ej apod, ejaculatory apodeme; goncx, gonocoxite; goncx apod, gonocoxal apodeme; gonst, gonostylus; hypd, hypandrium; ph, phallus. Scale bar = 0.1 mm.

Hilarimorphidae The Hilarimorphidae includes the single genus,Hilarimorpha Schiner. The male terminalia of Hilarimorpha (Figs 13 and 14) are characterized by the following derived features: subapical gonostyli (character 25); absence of lateral ejaculatory processes

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 392 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

(character 11:0); hypandrium and gonocoxites fused (character 16), with longitudinal ridge along midline (similar ridge present in many Bombyliidae, see Yeates 1994: character 111); ventral sheath of phallus extended anteriorly. In addition, the following plesiomorphic features characterize Hilarimorphidae: epandrium quadrate, not U-shaped; subepandrial region membranous. The phylogenetic affinities of this family remain ambiguous. It has been variously assigned as the sister group to the Bombyliidae, sister to Asiloidea, sister to Eremoneura (Trautwein et al. 2010), or sister to the Acroceridae (Wiegmann et al. 2011). Hilarimorpha and Apystomyia Melander are the only genera of the lower Brachycera to our knowledge that have the posterior end of the hypandrium invaginated to form a brace or apodeme supporting the position of the phallus ventrally (Figs 13 and 16, apod). In Hilarimorpha, the ventral sheath of the phallus articulates with this apodeme. In a sense, this brace could be interpreted as a precursor to the more anteriorly positioned phallapodeme (see below under “Apystomyiidae” and “Cyclorrhapha, exclusive of Opetiidae”).

‘Asiloidea’ + Eremoneura The monophyly of this lineage is supported on the basis of the gonostyli retracted to a subapical position on the gonocoxites (character 25) (Sinclair et al. 1994). Hilarimorphidae also have subapical gonostyli. Although the gonostyli are absent in Eremoneura, they are present in Apystomyiidae, probable sister to the Eremoneura (see below under “Apystomyiidae + Eremoneura”). The gonostyli articulate on the posterior margin of the gonocoxites in the therevoid family Evocoidae (Yeates et al. 2003), which is likely a reversal.

‘Asiloidea’ The definition and limits of this lineage are currently unclear. It has been considered to be monophyletic inclusive of Bombyliidae by Yeates (2002), monophyletic exclusive of Bombyliidae by Trautwein et al. (2010), and monophyletic exclusive of Hilarimorphidae by Wiegmann et al. (2011). In contrast, Woodley et al. (2009) showed Asiloidea (including Hilarimorphidae, Bombyliidae, Asilidae, Apioceridae, Mydidae, and the therevoid clade) to be paraphyletic in relation to the Eremoneura. No characters of the male terminalia are known that support the monophyly of Asiloidea exclusive of Eremoneura.

Asilidae + Apioceridae + Mydidae These three families are often considered to be most closely related, with the latter two families always indicated as sister groups (Dikow 2009a,b; Trautwein et al. 2010; Wiegmann et al. 2011). Characters of the male terminalia supporting this clade include the presence of muscle M38 (although also present in at least one genus of Therevidae) (Ovtshinnikova & Yeates 1998; Yeates 2002; Ovtshinnikova 2003), epandrium separated medially and joining proximally, and a number of non-genitalic characters

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 393

(Dikow 2009a,b). The epandrial character has not been used in analyses of higher classifications because it is considered rather homoplastic, found also in the therevoid clade and Eremoneura. Furthermore, coding and scoring this character is difficult due to variation in the division of the epandrium.

Asilidae The monophyly of this family is supported by five autapomorphies of the adult mouth- parts (Dikow 2009a). Male genitalic synapomorphies of the Asilidae include the base of the epandrium articulated with the anterodorsal prolongation of hypandrium (character 20:2) (Sinclair et al. 1994; Dikow 2009b), and a weakly sclerotized sperm sac (Dikow 2009b). The latter feature needs to be more clearly defined and evaluated outside the asiloid families. The so-called surstylus in Leptogastrinae (Asilidae) is not homologous with that in the Eremoneura. In these asilids, the epandrial lamella is characterized by a sub-basal notch, enabling each lobe to function together as claspers (Ovtshinnikova 1989; Dikow 2009a). The subepandrial sclerite is not fully developed or present on the inner margin of the epandrium (for discussion see Dikow 2009a: 74) and consequently the claspers are not homologous with surstyli.

Apioceridae The family Apioceridae, now restricted to the single genus Apiocera Westwood, is defined on the basis of a number of synapomorphies, including the phallus subdivided into a dorsal guide and a ventral needle-like component (character 26) (Sinclair et al. 1994; Yeates & Irwin 1996).

Mydidae The Mydidae are defined on the basis of a number of male genitalic synapomorphies, including the loss of lateral ejaculatory processes (character 11:0) and associated muscle M32, hypandrium fused to the gonocoxites (character 16), loss of gonostyli (character 23) and associated muscle M28 (Sinclair et al. 1994; Yeates & Irwin 1996; Ovtshinnikova 2003), and sperm chamber enclosed by the phallus (Dikow 2009a,b). The enclosure of the sperm chamber within the phallus in Mydidae requires better definition and comparison with other asiloid families.

Therevoid clade This group includes the families Apsilocephalidae, Evocoidae, Scenopinidae and Therevidae (Yeates et al. 2003; Winterton 2004; Trautwein et al. 2010). No male genitalic characters are known that support this clade.

Scenopinidae This family is currently defined on the basis of the following derived features within the therevoid clade: a deeply divided epandrium (also present in all Apioceridae, many

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 394 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Mydidae and some Asilidae) and a phallus with two or three tubes with corresponding phallotremata (Yeates 1992). Sinclair et al. (1994) hypothesized that the lateral ejaculatory processes were absent in Scenopinidae, but Ovtshinnikova & Yeates (1998) found on the basis of musculature that the processes were fused into a single apodeme dorsal to the ejaculatory apodeme in Metatrichia waterhousei (Paramonov).

Therevidae Although this family is rather weakly defined and lacks distinct morphological synapomorphies (Yeates 2002), its monophyly has been supported by molecular evidence (Yang et al. 2000; Trautwein et al. 2010). Sinclair et al. (1994) found that the lateral ejaculatory processes were either retracted into the base of the phallus (as supported by musculature in Ovtshinnikova 1989, fig. 179), absent, or positioned at the base of the phallus (Ovtshinnikova & Yeates 1998).

Apsilocephalidae As reviewed and illustrated in Sinclair et al. (1994, fig. 16a, b),Apsilocephala Kröber is a peculiar therevoid genus characterized by the presence of articulated surstyli. Along with two other genera, Apsilocephala was assigned to the family Apsilocephalidae, which was considered to be most closely related to the Empidoidea or Eremoneura due to the presence of surstyli (Nagatomi et al. 1991). However, both of the other included genera (Clesthentia White and Kaurimyia Winterton & Irwin) lack articulated later- oapical epandrial lobes and the subepandrial sclerite does not extend onto the apex of the epandrial lamellae. Consequently, it is apparent that surstyli are only present in Apsilocephala and we agree with Winterton & Irwin (2008) that this feature is not a defining character of the family, or a synapomorphy shared with Eremoneura. This family was assigned to the therevoid clade in analyses of Yeates (2002), Yang et al. (2000) and Wiegmann et al. (2011).

Apystomyiidae + Eremoneura The classification of the enigmatic monotypic genus Apystomyia has been questioned since its description more than 60 years ago (Melander 1950). Presently the genus is assigned to its own family, the Apistomyiidae (Nagatomi & Liu 1994). The phylogenetic history of Apystomyia was reviewed by Nagatomi & Liu (1994) and Trautwein et al. (2010) and based on nuclear genes CAD and 28S rDNA, it was considered by Trautwein et al. (2010) to be the sister group to the Cyclorrhapha. Morphological evidence assigns Apystomyiidae as the sister group to Eremoneura (rather than Cyclorrhapha) on the basis of the following synapomorphies of the male terminalia (Figs 15–19): lateral ejaculatory processes absent (character 11:0); hypandrium and gonocoxites fused (character 16); epandrium deeply U-shaped (character 27); subepandrial sclerite fully developed, sclerotized from base of hypoproct to base of phallus (character 28); paired lateral arms of the subepandrial sclerite (i.e., bacilliform sclerites) fused to the inner margin of the epandrium and extending to tips of the epandrium (character 29); surstyli functionally developed, but not articulated

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 395

(character 30:1). These synapomorphies combined with the lack of the three male genitalic synapomorphies that define the Eremoneura (see below), namely gonostyli absent (character 23), presence of postgonites (character 31) and a phallic plate (character 32), together indicate a sister group relationship between Apystomyiidae and Eremoneura. Several of the synapomorphies listed above are highly homoplastic (e.g., fusion of hypandrium and gonocoxites), but the development in Apystomyiidae of functional surstyli with complete subepandrial and bacilliform sclerites, while retaining gonostyli and lacking postgonites, provides strong support for this sister group relationship with Eremoneura.

Apystomyiidae The male and female terminalia ofApystomyia were illustrated by Wiegmann et al. (1993), Nagatomi & Liu (1994) and Yeates (1994). Several key features in the male terminalia were overlooked by these authors and for this reason we provide the following brief diagnosis of the male terminalia (Figs 15–19): Postabdomen and terminalia symmetrical, unrotated. Epandrium deeply U-shaped. Subepandrial sclerite fully developed, sclerotized from base of hypoproct to base of phallus; paired lateral bacilliform sclerites fused to inner margin of epandrium, extending to apex of epandrial lamella. Surstyli functionally developed, lacking basal articulation. Hypandrium and gonocoxites fused completely, processes of gonocoxal apodemes present, moderately extended anteriorly; gonostyli present, subapically position on gonocoxite, articulated obliquely; postgonites absent; posterior apex of hypandrium invaginated anteriorly forming apodeme supporting ventral aspect of phallus. Phallus short, stout with broad phallotrema; ejaculatory apodeme articulated at based of phallus, lever-like; sperm chamber large, membrane extending laterally beyond margin of phallus; ventral sheath of phallus prolonged anteriorly as pair of divergent apodemes; lateral ejaculatory processes absent. Apystomyia does not share synapomorphies with the Empidoidea. For example, females of Apystomyia have three spermathecae rather than the single synapomorphic spermatheca that is characteristic of Empidoidea (Sinclair & Cumming 2006). The similar lever-like ejaculatory apodeme observed in Apystomyia and Empidoidea (character 14:1) could be considered apomorphic and possibly due to homoplasy (as mapped in Figs 2 and 3). However, it is more likely that this feature is part of an ordered trans- formation series, beginning with the ejaculatory apodeme being attached to the anterior margin of sperm chamber (functioning through a piston-like contraction of the sperm chamber) (character 14:0), to being only ventrally articulated with the phallus (functioning as a lever to compress the sperm chamber) (character 14:1) and thus partially separated from it, to ending up being entirely separated from the phallus by the sperm duct (characteristic of Cyclorrhapha) (character 14:2). In addition, Apystomyia shares no synapomorphies with the Cyclorrhapha, including not pupating in the characteristic cyclorrhaphan puparium (as indicated indirectly by the presence of abdominal plaques in Apystomyia), not having circumverted 360° rotated male terminalia

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 396 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Figs 15–19. Apystomyia elingus Melander. (15) Terminalia, lateral view. (16) Hypandrium, gonostylus and phallus, lateral view. (17) Phallus and ejaculatory apodeme, lateral view. (18) Epandrium and proctiger, ventral view. (19) Hypandrium, gonostylus and phallus, dorsal view. Abbreviations: apod, apodeme; bac scl, bacilliform sclerite; cerc, cercus; ej apod, ejaculatory apodeme; ej duct, ejaculatory duct; epand, epandrium; goncx, gonocoxite; goncx apod, gonocoxal apodeme; gonst, gonostylus; hypd, hypandrium; hyprct, hypoproct; ph, phallus; sbepand scl, subepandrial sclerite; spm chmbr, sperm chamber; sur, surstylus. Scale bar = 0.1 mm.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 397

(character 34:0 rather than 34:3), retaining processes of the gonocoxal apodemes (character 19:0 rather than 19:2), and lacking the characteristic cyclorrhaphan arista observed in Cyclorrhapha exclusive of Opetiidae. The only derived feature that Apystomyia potentially shares with Cyclorrhapha is a possible precursor of the phallapodeme (see below under “Cyclorrhapha, exclusive of Opetiidae”). There is a short invagination from the posterior margin of the hypandrium to the mid ventral margin of the phallus that appears to form an apodeme supporting the phallus ventrally (Figs 15, 16, apod). A similar slender invaginated apodeme is present in Hilarimorpha (Fig. 13, apod), but the apodeme extends to the anteroventral margin of the phallus.

Eremoneura The monophyly of the Eremoneura is supported on the basis of numerous derived characters (Cumming et al. 1995; Woodley et al. 2009; Lambkin et al. 2013). The following are two eremoneuran autoapomorphies of the male terminalia (Fig. 2): gonostylus absent (character 23); postgonite present (character 31) (see Sinclair 2000 and Sinclair & Cumming 2006 for discussion of the homology of these eremoneuran features). The presence of the phallic plate (character 32) is tentatively proposed as an additional synapomorphy. The phallic plate is a short to long sclerotization of the base of the phallus and hypandrial arms towards the base of the subepandrial sclerite (see Sinclair & Cumming 2006, fig. 280) that possibly is part of the ground plan of Eremoneura, although it is not well developed in some lineages and needs further study. The phallic plate has been identified in Empididae, Nerioidea and Diopsoidea (Sinclair 2000; Shatalkin 1995), where it may have evolved independently.

Empidoidea The monophyly of the Empidoidea (Fig. 3) is supported by all recent studies (Collins & Wiegmann 2002a; Sinclair & Cumming 2006; Moulton & Wiegmann 2007; Wiegmann et al. 2011). A single derived character of the male terminalia supports the Empidoidea: ejaculatory apodeme ventrally articulated and lever-like (character 14:1) (Cumming et al. 1995). However, a similar ventrally articulated ejaculatory apodeme is newly recognized in Apystomyiidae, which may be homologous and part of a trans- formation series leading to the separated sperm pump in Cyclorrhapha (character 14:2) (see discussion above under “Apystomyiidae”). The Atelestidae + Hybotidae have been considered a monophyletic group on the basis of a number of morphological characters including the ejaculatory apodeme fused to the phallus (character 14:4) (Sinclair & Cumming 2006). However, within the Atelestidae, this fusion of the ejaculatory apodeme with the phallus occurs only in the genus Atelestus Walker (Sinclair & Kirk-Spriggs 2010). In molecular studies, the Atelestidae are assigned as the sister to the remaining Empidoidea (Collins & Wiegmann 2002a; Moulton & Wiegmann 2007; Wiegmann et al. 2011).

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 398 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Atelestidae The monophyly of the Atelestidae (including Nemedininae) is supported by greatly lengthened processes of the gonocoxal apodemes (character 19:1) and a shortened hypandrium (character 33) (Cumming et al. 1995; Sinclair & Cumming 2006; Sinclair & Kirk-Spriggs 2010). The presence of subapical surstyli is considered a synapomorphy of the Atelestinae.

Hybotidae The Hybotidae are a well-defined family, supported by the following characters of the male terminalia: gonocoxal apodeme confined to anterior margin of hypandrium, lacking narrow process (character 19:2); male terminalia rotated between 45-90° to the right (character 34:1) (Cumming et al. 1995; Sinclair & Cumming 2006). Although the male terminalia of hybotids are often strongly asymmetrical, asymmetry is possibly not a ground plan character of the family, given the symmetrical condition found in the apparently plesiomorphic genus Trichinomyia Tuomikoski (see Cumming et al. 1995, fig. 12; Sinclair & Cumming 2006).

Dolichopodidae s.lat. The monophyly of the Dolichopodidae (inclusive of Microphorinae and Parath alassiinae) is supported by the following characters of the male terminalia: gonocoxal apodeme confined to anterior margin of hypandrium, lacking narrow process (character 19:2); male genitalia rotated and lateroflexed to the right (resulting in functional circumversion) (character 34:2); bacilliform sclerites fused to hypandrium, separated from base of phallus (character 35); and male terminalia asymmetrical around left-side displaced foramen (character 36, less pronounced in Microphorinae and Parath alassiinae) (Cumming et al. 1995; Sinclair & Cumming 2006).

Cyclorrhapha This is an extremely well-supported clade (Fig. 3), corroborated by the following characters of the male terminalia: ejaculatory apodeme and sperm pump separated from phallus by short to long sperm duct (character 14:2); gonocoxal apodeme confined to anterior margin of hypandrium, lacking narrow process (character 19:2); surstyli articulated (character 30:2; this feature has also independently evolved numerous times in Empidoidea); and terminalia circumverted, rotated 360° (character 34:3) (Cumming et al. 1995; Sinclair & Cumming 2006; Lambkin et al. 2013). Reduction of the process of the gonocoxal apodemes in Cyclorrhapha appears to have evolved to compensate for the circumverted 360° rotation of the terminalia that characterizes this lineage, including Opetiidae. A similar reduction is observed in other groups within the lower Eremoneura that exhibit permanent male terminalia rotation (Cumming et al. 1995). Similarly, the separated sperm pump in Cyclorrhapha,

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 399 including Opetiidae, is possibly a consequence of circumversion. However, the functionally circumverted condition seen in Dolichopodidae s.lat., which is obtained through a dextral rotation and lateroflexion to the right side of the postabdomen (character 34:2), appears to have evolved with the sperm pump still ventrally attached to the phallus.

Cyclorrhapha, exclusive of Opetiidae The phallapodeme is present as a longitudinal invagination of the hypandrium that surrounds the base of the phallus in all cyclorrhaphan families except Opetiidae (character 37) (Cumming et al. 1995, figs 17–20). A possible precursor to this condition is a newly identified, somewhat more posteriorly positioned invagination in Apystomyia and Hilarimorpha that apparently forms a brace for the phallus ventrally (Figs 13, 15 and 16; see above under “Hilarimorphidae” and “Apystomyiidae”), but its homology requires further investigation (i.e., study of musculature). Several additional non- genitalic characters supporting a sister group relationship between Opetiidae and the remainder of the Cyclorrhapha are listed by Cumming et al. (1995, characters R, S, T). A number of recent studies propose alternate classifications. Opetiidae were assigned as sister to the Platypezidae and Phoroidea by Wiegmann et al. (2011), and Lonchopteridae were assigned as sister to the remaining Cyclorrhapha on the basis of larval characters (Rotheray & Gilbert 2008).

Lonchopteridae + Phoroidea This lineage is supported by several non-genitalic characters (Cumming et al. 1995, characters Y, Z, AA) and the following male postabdominal character: segment 7 with no distinguishable sclerite (characters 38), although spiracle 7 remains present in Lonchoptera uniseta Curran and Sciadocerinae (Hennig 1976; Cumming et al. 1995; McAlpine 2008). The Lonchopteridae were assigned as the sister to the remaining families of the Platypezoidea (Opetiidae, Platypezidae, Ironomyiidae and Phoridae) in Wiegmann et al. (2011).

Lonchopteridae The Lonchopteridae are a small and distinctive family, with unique larvae and odd wing venation. Two characters of the male terminalia define the family: subepandrial sclerite greatly reduced (character 28:2) (Hennig 1976, figs 5 and 7, labelled 10. St); surstyli absent (character 30:0) (Cumming et al. 1995; Sinclair & Cumming 2006).

Ironomyiidae + Phoridae A single male terminalia character supports this clade: postgonites absent (character 31:0) (Cumming et al. 1995).

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 400 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Ironomyiidae The Ironomyiidae comprise a single extant genus and three species (McAlpine 2008). In addition to apomorphic characters of the antenna, this family is defined by the fusion or union of sternite 8 to the epandrium (character 39) and an articulated phallapodeme (character 40) (see below under “Syrphoidea + Schizophora”) (Cumming et al. 1995; McAlpine 2008).

Phoridae The ground plan condition of the male terminalia of the Phoridae was outlined by Nakayama (2012) and includes a four-point clasping motion (right and left surstylus and both sides of posterior margin of epandrium). Burmophora Beyer, a genus of the subfamily Hypocerinae, is considered by Nakayama (2012) to best illustrate the ground plan condition of the Phoridae. In this genus the male terminalia are nearly symmetrical and possess a pair of articulated surstyli. The male terminalia of the subfamily Sciadocerinae, considered to be the sister group to the rest of the family (Disney 2001), are autapomorphic and do not represent the plesiomorphic ground plan state of the Phoridae (Nakayama 2012).

Syrphoidea + Schizophora The families Syrphidae and Pipunculidae together with Schizophora, are now generally accepted as forming a monophyletic group (Woodley et al. 2009; Wiegmann et al. 2011). This relationship is also supported by a single character of the male terminalia: phallapodeme articulated (character 40), although it is also articulated in Ironomyiidae and several species of Lonchopteridae (Cumming et al. 1995).

Syrphoidea The Syrphidae and Pipunculidae have been regarded as a monophyletic group, based primarily on the male terminalia being deflexed under the right side of the abdomen (character 41) (Griffiths 1972; Cumming et al. 1995; Woodley et al. 2009). In several molecular studies, however, the monophyly of the Syrphoidea is contradicted with the Pipunculidae alone assigned as sister to the Schizophora (Collins & Wiegmann 2002b; Moulton & Wiegmann 2007; Wiegmann et al. 2011).

Schizophora The monophyly of the Schizophora is supported by the fundamental autapomorphic development of the ptilinum with an externally visible ptilinal suture (Woodley et al. 2009). In addition, the following synapomorphies of the male terminalia of Schizophora have been proposed: postgonites adducted against the hypandrium (character 42); circumversion completed entirely within the puparium (character 43) (Cumming et al. 1995). In many non-schizophoran Cyclorrhapha the first 180° of circumversion occurs within the puparium (incomplete circumversion) whereas the remaining 180° of

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 401 rotation is completed immediately after eclosion and is partially reversible during mating (see discussion in Cumming et al. 1995, p. 136). Circumversion completed within the puparium, as in Schizophora, is non-reversible and allows for an upright (male above female) mating position (McAlpine 1981). Unfortunately information on the process of circumversion is lacking in some critical taxa. Cumming et al. (1995) indicated that incomplete circumversion has been observed (or inferred) in at least Opetiidae, Platypezidae, Phoridae, and Syrphidae. Circumversion completed within the puparium is reported for non-Schizophora in at least Ironomyia White (Ironomyiidae), Nephrocerus Zetterstedt (Pipunculidae) and Lonchoptera uniseta Curran (Lonchopteridae) based on fusion of sternite 8 with the epandrium (tergite 9), which requires complete rotation of the hypopygial tissue within the puparium before the fused sclerites form (Cumming et al. 1995). A more thorough survey of the process of circumversion is required, which could possibly help resolve the competing sister group relationships with Schizophora (see above under “Syrphoidea”). As reviewed in Woodley et al. (2009) the enormous diversity and size of the Schizophora has prevented cladistic analyses of the entire group. The study of McAlpine (1989) was the first broad attempt to cladistically analyze the group, and the paper by Wiegmann et al. (2011) was the first quantitative study of schizophoran phylogeny. Several of McAlpine’s (1989) superfamilies were recovered by Wiegmann et al. (2011), but the assignment of a large number of families remained unresolved and relationships between most superfamilies were weak (Fig. 4).

Tephritoidea In a morphological analysis of the Tephritoidea, Korneyev (1999) indicated the following derived characters of the male terminalia: phallus elongate, coiled (character 44:3), and prensisetae present (character 45) (similar peg-like setae are present in Drosophilidae, Grimaldi 1990). Within this superfamily, the families Platystomatidae, Ctenostylidae, Tephritidae Pyrgotidae and Tachiniscidae (i.e., higher Tephritoidea) are united on the basis of a discrete structure on the apex of the distiphallus, termed the glans (character 44:2), and the presence of the fultelliform sclerite (or phallic guide) joined to the phallapodeme (character 46) (McAlpine 1989; Korneyev 1999).

Nerioidea This superfamily is characterized by a number of autapomorphies of the male terminalia including: hypandrium reduced (character 33); elongate phallus directed anteriorly (character 44:4); elongate pregonites (character 47); elongate phallic plate (character 48) (Griffiths 1972; McAlpine 1989; McAlpine 1996; Sinclair 2000).

Carnoidea According to Buck (2006) the following autapomorphy supports the monophyly of this superfamily: distiphallus flexible and slender (more or less ribbon-shaped), largely

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 402 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 or completely desclerotized and microtrichose (character 44:1). The distiphallus is further modified within this superfamily in Chloropidae + Milichiidae, becoming membranous and truncate (Brake 2000; Buck 2006).

Lauxanioidea The monophyly of this superfamily is reasonably well supported including the following autapomorphy of the postabdomen: a single dorsal sclerite, nearly symmetrical between 6th tergite and terminalia (possibly fusion of sternites 7 and 8) (character 49) (Griffiths 1972; McAlpine 1989). The Lauxaniidae are the most diverse included family, whose monophyly is supported by the repeatedly branched male accessory gland, forming a dense tangle (Sturtevant 1926).

Sciomyzoidea This superfamily is characterized by a relatively reduced and shortened male tergite 6 (character 50) (Griffiths 1972; McAlpine 1989), although McAlpine (1991) considered this character to have only been derived within a large subgroup of the superfamily.

Ephydroidea Griffiths (1972), Chandler (1987) and McAlpine (1989) listed the loss of the sclerites of abdominal segment 8 (character 51) and a short phallus, as autapomorphies of the Ephydroidea. In addition, male sternite 7 is lost in all included families except Curtonotidae (character 52) (Chandler 1987; Grimaldi 1990), although sensilla associated with sternite 7 remain in some Drosophilidae (Marshall 1960). The definition of the shortened phallus is rather vague and requires additional coding and refinement.

Calyptratae The Calyptratae are currently classified into the Hippoboscoidea, Muscoidea and Oestroidea (McAlpine 1989). The monophyly of this huge lineage is supported by the molecular analyses of Kutty et al. (2010) and Wiegmann et al. (2011), and several morphological autapomorphies (Griffiths 1972; McAlpine 1989). Despite this, we have found no convincing characters of the male terminalia of Calyptratae that define the entire clade. Griffiths (1972) proposed that members of the Calyptratae share the following synapomorphy of the male terminalia: the posterior dorsal displacement of the 7th left spiracle onto the sclerite of syntergosternite 7 + 8. This modification needs to be thor- oughly surveyed as it occurs sporadically elsewhere in the Cyclorrhapha and exhibits some variability within Calyptratae. For example within Hippoboscoidea the 7th left spiracle is displaced in Glossina Wiedemann (Griffiths 1972, fig. 67; Pollock 1973,

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 403 fig. 11), but both spiracles lie in the membrane in Hippobosca L. (see Maa & Peterson 1987, fig. 33). In addition, in the oestroid genus Gastrophilus Leach the spiracles of the 7th segment lie in the membrane in G. pecorum Fabricius, whereas the left spiracle is in the derived position in G. intestinalis De Geer (Pollock 1973, figs 1 and 6). The Hippoboscoidea and Oestroidea are considered monophyletic superfamilies on the basis of both morphological and molecular analyses (McAlpine 1989; Wiegmann et al. 2011). However, the Muscoidea (i.e., Fanniidae, Muscidae, Anthomyiidae, Scathophagidae) have been found to be paraphyletic in relation to the Oestroidea (Kutty et al. 2008). The morphological synapomorphies of the male terminalia that McAlpine (1989) listed in support of the Muscoidea are rather weak and homoplastic.

Hippoboscoidea This is a distinctive group characterized by development of larvae within the female oviduct (McAlpine 1989; Meier et al. 1999). Petersen et al. (2007) analyzed family relationships within the Hippoboscoidea using molecular data. Among features of the male terminalia, the superfamily has an ejaculatory apodeme that is lost or desclerotized (character 9) and cerci that are reduced (character 53) (Griffiths 1972; Pollock 1973; McAlpine 1989).

Fanniidae The monophyly of the Fanniidae is well characterized and includes the following characters of the male terminalia: ejaculatory apodeme absent (character 9); sclerites of segment 6-8 fused, symmetrical and containing two pairs of spiracles (character 54) (Griffiths 1972). In addition, McAlpine (1989) listed bacilliform sclerites bearing processes, and a reduced phallus as synapomorphies of the family, but these are restricted to species groups of Fannia Robineau-Desvoidy and are not present in other fanniid genera (Chillcott 1960).

Muscidae + Anthomyiidae + Scathophagidae + Oestroidea The presence of the epiphallus was originally proposed as a synapomorphy of the Calyptratae, though it is absent in Fanniidae and Hippoboscoidea (Griffiths 1972). When traced on the cladogram (Fig. 4), the presence of the epiphallus (character 55) is assigned as a synapomorphy of Muscidae + Anthomyiidae + Scathophagidae + Oestroidea.

Oestroidea There is apparently no strong consensus about male terminalia synapomorphies supporting the entire Oestroidea. In addition to Wiegmann et al. (2011), the family relationships within the Oestroidea have been addressed in other analyses of molecular data by Kutty et al. (2008, 2010) and Marinho et al. (2012). The presence of a postgonal apodeme (character 56) and a phallus with denticles on the ventral surface (character 57) have been recognized as synapomorphies of the superfamily (McAlpine 1989; Pape 1992). Lambkin et al. (2013) listed the former character, as well as the dorsal wall

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 404 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 of distiphallic tube sclerotized and forming the dorsal plate (character 58), and the presence of a mesohypophallic sclerotization mid-ventrally along the distiphallus (character 59), as synapomorphies of Oestroidea. The latter character is lost secondar- ily in Oestridae, Sarcophagidae and Tachinidae, and the postgonal apodeme is absent in axiniids and mystacinobines (Rognes 1997), but see updates to this character distri- bution in Pape & Arnaud (2001). Within the Oestroidea, fusion of dorsolateral phallic processes are considered synapomorphic for the Tachinidae + Sarcophagidae (Pape 1992), and reduction of the bacilliform sclerites is probably autapomorphic for Sarcophagidae (Pape 1992).

Discussion Male terminalia are an extremely important source of phylogenetic information within Diptera at all taxonomic levels. This is because the character system exhibits tremen- dous morphological diversity associated with sexual selection resulting from female choice (Eberhard 1985, 2004), and displays suites of characters correlated with evolutionary changes linked to the process of sperm transfer (Sinclair et al. 2007) as well as adaptive shifts in mating position (Cumming et al. 1995; Huber et al. 2007). Sperm is discharged directly into the female by the sperm pump or a muscular ejaculatory duct in the ground plan of Diptera. A sperm pump with a functional ejaculatory apodeme is a possible synapomorphy of all Diptera, exclusive of Nymphomyiidae and Deuterophlebiidae. Production of spermatophores in the lower Diptera is associated with the independent development of a multi-chambered accessory gland (character 6) in Ptychopteridae, Culicomorpha and Bibionomorpha s.str. This is correlated with desclerotization of the aedeagus (character 8) and loss of the ejaculatory apodeme (character 9) in Culicomorpha. However, formation and transfer of spermatophores in Bibionomorpha s.str. is enabled by a modified ventrally positioned ejaculatory apodeme (character 10) that appears to push out the spermatophore as it forms. This lineage also has a partially desclerotized aedeagus that is encircled by a parameral sheath (character 5), which appears to be a synapomorphy of the exceedingly diverse Neodiptera, exclusive of the family Axymyiidae (Fig. 1). Shifts in mating positions from tail-to-tail configurations to male-dominated mating stances, with the male above female, have led to a number of apomorphic changes in the male terminalia throughout the order. These shifts in mating position have occurred independently in various groups in the nematocerous Diptera and in the lower Brachycera, resulting in differing degrees of facultative or permanent rotation in the male terminalia of these groups. For example, in many nematocerous Diptera and lower Brachycera, such as certain genera of Tipulidae, Chironomidae, Ceratopogonidae, Bombyliidae and Asilidae, the male genitalia become temporarily inverted (rotated from 90° to 180°) during mating (McAlpine 1981; Bickel 1990). In other groups such as Culicidae, inversion is permanent, occurring after emergence but before mating (McAlpine 1981). Permanent rotation of male terminalia within much of the Muscomorpha, appears to have led to the evolution of numerous genitalic

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 405 apomorphies and is probably one of the reasons that this character system has been so informative at this level in the Brachycera (Fig. 2). This includes reduction of the processes of the gonocoxal apodemes (character 19:2) and development of asymmetries within Eremoneura (characters 34, 36, 41), functional circumversion in Dolichopodidae s.lat. (character 34:2), as well as separation of the sperm pump (character 14:2) and complete circumversion (character 34:3) in the Cyclorrhapha (Fig. 3). The change from ventral clasping in the lower Diptera to dorsal clasping in Eremoneura (characters 23 and 27–30) may also be predominantly correlated with shifts in mating position and genitalic rotation. Recent molecular studies have made very important contributions towards the latest advances in Diptera phylogenetics (Yeates & Wiegmann 2012). Although the molecular based cladograms presented by Wiegmann et al. (2011) have been helpful in tracing character evolution in the male terminalia of Diptera, several incongruences are apparent. For example, their placement of Nemestrinidae as the sister group to Xylophagidae is refuted by several synapomorphies of the male terminalia of the Muscomorpha (characters 2:0, 20–22) to which the Nemestrinidae belong, as well as the absence of an endoaedeagus (character 15), which is a newly interpreted synapomorphy for Xylophagomorpha + Tabanomorpha (Fig. 2). The assignment of Apystomyiidae as the sister group to the Cyclorrhapha by Trautwein et al. (2010) and Wiegmann et al. (2011) based on molecular data is also controversial. However, on the basis of four synapomorphies of the male terminalia (characters 27–30) in conjunction with the lack of three eremoneuran synapomorphies (characters 23, 31 and 32), it appears more reasonable to consider Apystomyiidae to be the sister group to the Eremoneura (Fig. 2). Further study of this interesting family is required. There is certainly a need to search for new genes and other molecular characters. Many recent molecular papers are the result of analyses of similar or identical gene sequences, often generated by the same laboratory or collaborative team (e.g., Wiegmann et al. 2000, 2011; Yang et al. 2000; Collins & Wiegmann 2002a,b; Moulton & Wiegmann 2007; Petersen et al. 2007; Yeates et al. 2007; Bertone et al. 2008; Kutty et al. 2008, 2010; Peterson et al. 2010; Trautwein et al. 2010, 2011). We encourage total evidence analyses of combined molecular and morphological data modelled after Dikow (2009b), who analyzed and compared the contributions of the data sets on the resulting cladograms of Asilidae. Additional investigation of complex morphological character systems such as the male terminalia of Diptera will continue to assist with the resolution of phylogenetic relationships within the order. This is especially true for the lower Diptera and Schizophora, where hypotheses of relationship appear somewhat unstable, or for the latter group, extremely preliminary.

Acknowledgements We extend our sincere thanks and appreciation to Monty Wood who encouraged us to take on the study of male genitalic homologies. Our Diptera colleagues (Jim O’Hara,

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 406 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Jeff Skevington and Owen Lonsdale) at the Canadian National Collection of Insects are also thanked for their guidance and helpful discussions.

References Abul-Nasr, S.E. (1950) Structure and development of the reproductive system of some species of Nematocera (Order Diptera: Suborder Nematocera). Philosophical Transactions of the Royal Society of London, Ser. B, Biological Sciences 234: 339–396. Amorim, D.S. (1993) A phylogenetic analysis of the basal groups of Bibionomorpha, with a critical reanalysis of the wing vein homology. Revista brasileira de Biologia 52: 379–399. Amorim, D.S. & Grimaldi, D.A. (2006) Valeseguyidae, a new family of Diptera in the Scatopsoidea, with a new genus in Cretaceous amber from Myanmar. Systematic Entomology 31: 508–516. Bertone, M.A., Courtney, C.W. & Wiegmann, B.M. (2008) Phylogenetics and temporal diversification of the earliest true flies (Insecta: Diptera) based on multiple nuclear genes. Systematic Entomology 33: 668–687. Beutel, R.G., Friedrich, F., Hörnschemeyer, T., Pohl, H., Hünefeld, F., Beckmann, F., Meier, R., Misof, B., Whiting, M.F. & Vilhelmsen, L. (2011) Morphological and molecular evidence converge upon a robust phylogeny of the megadiverse Holometabola. Cladistics 27: 341–355. Bickel, D.J. (1990) Sex with a twist in the tail. New Scientist 1731: 34–37. Blaschke-Berthold, U. (1994) Anatomie und Phylogenie der Bibionomorpha (Insecta, Diptera). Bonner Zoologische Monographien 34: 1–206. Black, V.H. (1966) The pupal development of the male genital disc of Eucalliphora lilaea, Unpublished M.A. thesis, Sacramento State College, Sacramento, CA, 40 pp. Borkent, A. (2012) The pupae of Culicomorpha—morphology and a new phylogenetic tree. Zootaxa 3396: 1–98. Borkent, A., Borkent, C.J. & Sinclair, B.J. (2008) The male genital tract of Chaoboridae (Diptera: Culicomorpha). The Canadian Entomologist 140: 621–629. Borkent, A. & Sinclair, B.J. (2012) The male genital tract of Axymyiidae and Tanyderidae (Diptera). In Festschrift commemorating the coordinators of the Manual of Nearctic Diptera and their contributions to building the Canadian National Collection of Insects. Part 3. The Canadian Entomologist 144: 266–272. Brake, I. (2000) Phylogenetic systematics of the Milichiidae (Diptera, Schizophora). Entomologica Scandinavica Supplementum No. 57, 120 pp. Buck, M. (2006) A new family and genus of acalypterate flies from the Neotropical region, with a phylogenetic analysis of Carnoidea family relationships (Diptera, Schizophora). Systematic Entomology 31: 377–404. Cannings, R.A. (2002) The systematics of Lasiopogon (Diptera: Asilidae). Royal British Columbia Museum, Victoria, BC, vi + 354 pp. Chandler, P.J. (1987) The families Diastatidae and Campichoetidae (Diptera, Drosophilidae) with a revision of Palearctic and Nepalese species of Diastata Meigen. Entomologica Scandinavica 18: 1–50. Chillcott, J.G. (1960) A revision of the Nearctic species of Fanniinae (Diptera: Muscidae). The Canadian Entomologist. Supplement 14, 295 pp. Cole, F.R. (1927) A study of the terminal abdominal structures of male Diptera (two-winged flies). Proceedings of the California Academy of Sciences, 4th series. 16(14): 397–499. Colless, D.H. (1969) The genus Perissomma (Diptera: Perissommatidae), with new species from Australia and Chile. Australian Journal of Zoology 17: 719–728. Collins, K.P. & Wiegmann, B.M. (2002a) Phylogenetic relationships and placement of the Empidoidea (Diptera: Brachycera) based on 28S rDNA and EF-1a sequences. Insect Systematics & Evolution 33: 421–444. Collins, K.P. & Wiegmann, B.M. (2002b) Phylogenetic relationships of the lower Cyclorrhapha (Diptera: Brachycera) based on 28S rDNA sequences. Insect Systematics & Evolution 33: 445–456.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 407

Cook, E.F. (1981) Scatopsidae [Chapter] 20. In: McAlpine, J.F., Peterson, B.V., Shewell, G.E., Teskey, H.J., Vockeroth, J.R. & Wood, D.M. (co-ords.) Manual of Nearctic Diptera, Volume 1. Agriculture Canada Monograph 27: 313–319. Courtney, G.W. (1990) Revision of Nearctic mountain midges (Diptera: Deuterophlebiidae). Journal of Natural History 24: 81–118. Courtney, G.W. (1994) Biosystematics of the Nymphomyiidae (Insecta: Diptera): life history, morphology, and phylogenetic relationships. Smithsonian Contributions to Zoology No. 550, 41 pp. Crampton, G.C. (1936) Suggestions for a new interpretation of the postabdomen in male Cyclorrhaphous Diptera. Bulletin of the Brooklyn Entomological Society 31: 141–149. Crampton, G.C. (1942) The external morphology of the Diptera. In: Guide to the Insects of Connecticut. Part VI. The Diptera or true flies of Connecticut.Bulletin of the Connecticut State Geological and Natural History Survey 64: 10–165. Crampton, G.C. (1944) A comparative morphological study of the terminalia of male calypterate cyclorrhaphous Diptera and their acalyptrate relatives. Bulletin of the Brooklyn Entomological Society 39: 1–31. Cumming, J.M. & Sinclair, B.J. (1996) The higher-level phylogeny of Eremoneura (Diptera: Brachycera). Proceedings of the XX International Congress of Entomology. Florence, Italy: p. 24. Cumming, J.M., Sinclair, B.J., Brooks, S.E., O’Hara, J.E. & Skevington, J.H. (2011) The history of dipterology at the Canadian National Collection of Insects, with special reference to the Manual of Nearctic Diptera. In: Festschrift commemorating the coordinators of the Manual of Nearctic Diptera and their contributions to building the Canadian National Collection of Insects. Part 1. The Canadian Entomologist 143: 539–577. Cumming, J.M. & Sinclair, B.J. & Wood, D.M. (1995) Homology and phylogenetic implications of male genitalia in Diptera – Eremoneura. Entomologica Scandinavica 26: 121–151. Cumming, J.M. & Wood, D.M. (2009) Adult morphology and terminology. In: Brown, B.V., Borkent, A., Cumming, J.M., Wood, D.M. & Zumbado, M. (Eds) Manual of Central American Diptera. Volume 1. NRC Research Press, Ottawa, ON, pp. 9–50. Curler, G.R. & Moulton, J.K. (2012) Phylogeny of psychodid subfamilies (Diptera: Psychodidae) inferred from nuclear DNA sequences with a review of morphological evidence for relationships. Systematic Entomology 37: 603–616. Dahl, C. (1980) Comparison of postembryonic organization of the genital segments in Trichoceridae, Tipulidae, and Anisopodidae (Diptera, Nematocera). Zoologica Scripta 9: 165–185. Dikow, T. (2009a) Phylogeny of Asilidae inferred from morphological characters of imagines (Insecta: Diptera: Brachycera: Asiloidea). Bulletin of the American Museum of Natural History No. 319, 175 pp. Dikow, T. (2009b) A phylogenetic hypothesis for Asilidae based on a total evidence analysis of morphological and DNA sequence data (Insecta: Diptera: Brachycera: Asiloidea). Organisms, Diversity & Evolution 9: 165–188. Disney, R.H.L. (2001) Sciadoceridae (Diptera) reconsidered. Fragmenta faunistica 44: 309–317. Dobzhansky, T. (1930) Studies on the intersexes and supersexes in Drosophila melanogaster. Izvestiya Byuro po genetike, Akademiya Nauk SSSR 8: 91–155 [In Russian]. Dufour, L. (1851) Recherches anatomiques et physiologiques sur les Diptères. Mémoires présentés par divers savants à l’Académie des Sciences de l’Institut de France et imprimés par son ordre: Sciences mathé- matiques et physiques. Paris, 11: 171–360, pls. 1–11. Eberhard, W.G. (1985) Sexual selection and animal genitalia. Harvard University Press, Cambridge, MA, 244 pp. Eberhard, W.G. (2004) Male–female confict and genitalia: failure to confirm predictions in insects and spiders. Biological Reviews 79: 121–186. Emden, F.I. van & Hennig, W. (1956) Diptera. In: Tuxen, S.L. (ed.) Taxonomist’s glossary of genitalia of insects. Munksgaard, Copenhagen: pp. 111–122. Feuerborn, H.J. (1922) Das Hypopygium “inversum” und “circumversum” der Dipteren. Zoologischer Anzeiger 60: 189–212.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 408 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Fitzgerald, S.J. (2004) Evolution and classification of Bibionidae (Diptera: Bibionomorpha). Unpublished Ph.D. thesis, Oregon State University, Corvallis, OR, 385 pp. Frommer, S.I. (1963) Gross morphological studies of the reproduction system in representative North American crane flies (Diptera: Tipulidae). University of Kansas Science Bulletin 44: 535–626. Griffiths, G.C.D. (1972) The phylogenetic classification of Diptera Cyclorrhapha, with special reference to the structure of the male postabdomen. Series entomologica 8. Dr. W. Junk, The Hague, 340 pp. Griffiths, G.C.D. (1981) Book review. Manual of Nearctic Diptera. Volume 1. Bulletin of the Entomological Society of Canada 13: 49–55. Griffiths, G.C.D. (1990a) Book review. Manual of Nearctic Diptera. Volume 3. Quaestiones Entomologicae 26: 117–130. Griffiths, G.C.D. (1990b) Functional continuity in the male genitalia of Brachycera, the basic assumption of the periandrial theory. Second International Congress of Dipterology, Abstract Volume (Third Supplement). Bratislava, Czechoslovakia: p. 343. Griffiths, G.C.D. (1996) Review of papers on the male genitalia of Diptera by D.M. Wood and associates. Studia dipterologica 3: 107–123. Grimaldi, D.A. (1990) A phylogenetic, revised classification of genera in the Drosophilidae (Diptera). Bulletin of the American Museum of Natural History No. 197, 139 pp. Hennig, W. (1936) Beiträge zur Kenntinis des Kopulationsapparates der cyclorrhaphen Dipteren. Zeitschrift für Morphologie und Ökologie der Tiere 31: 328–370. Hennig, W. (1958) Die Familien der Diptera Schizophora und ihre phylogenetischen Verwandtschaftsbezichungen. Beiträge zur Entomologie 8(5/6): 505–688. Hennig, W. (1973) Diptera (Zweiflügler). Handbuch der Zoologie 4(2) 2/31: 1–337. Hennig, W. (1976) Das Hypopygium von Lonchoptera lutea Panzer und die phylogenetischen Verwandtschftsbeziehungen der Cyclorrphapha (Diptera). Stuttgarter Beiträge zur Naturkunde Serie A 283: 1–63. Hennig, W. (1981) Insect phylogeny. Translated and edited by A.C. Pont (revisionary notes by D. Schlee). John Wiley & Sons, New York, NY, 514 pp. Huber, B.A., Sinclair, B.J. & Schmitt, M. (2007) The evolution of asymmetric genitalia in spiders and insects. Biological Reviews 82: 647–698. Hünefeld, F. & Beutel, R.G. (2005) The sperm pumps of Strepsiptera and Antliophora (Hexapoda). Journal of Zoological Systematics and Evolutionary Research 43(4): 297–306. Kerr, P.H. (2010) Phylogeny and classification of Rhagionidae, with implications for Tabanomorpha (Diptera: Brachycera). Zootaxa 2592: 1–133. Korneyev, V.A. (1999) Phylogenetic relationships among the families of the superfamily Tephritoidea. In: Aluja, M. & Norrbom, A.L. (eds.) Fruit flies (Tephritidae): phylogeny and evolution of behavior. CRC Press, Boca Raton, FL, pp. 3–22. Krivosheina, N.P. (2012) The position of the genusHeterotropus in the Brachycera-Orthorrhapha (Diptera) system. Zoologicheskii Zhurnal 91: 337–346 [In Russian]. Kutty, S.N., Pape, T., Pont, A.C., Wiegmann, B.M. & Meier, R. (2008) The Muscoidea (Diptera: Calyptratae) are paraphyletic: Evidence from four mitochondrial and four nuclear genes. Molecular Phylogeny and Evolution 49: 639–652. Kutty, S.N., Pape, T., Wiegmann, B.M. & Meier, R. (2010) Molecular phylogeny of the Calyptratae (Diptera: Cyclorrhapha) with an emphasis on the superfamily Oestroidea and the position of Mystacinobiidae and McAlpine’s fly. Systematic Entomology 35: 614–635. Lambkin, C.L., Sinclair, B.J., Pape, T., Courtney, G.W., Skevington, J.H., Meier, R., Yeates, D.K., Blagoderov, V. & Wiegmann, B.M. (2013) The phylogenetic relationships among infraorders and superfamilies of Diptera based on morphological evidence. Systematic Entomology. 8: 164–179 Laugé, G. (1968) Morphologie comparée de la région génitale des intersexués triploïdes de Drosophila melanogaster. Annales de la Société Entomologique de France 4: 481–499. Maa, T.C. & Peterson, B.V. (1987) Hippoboscidae [Chapter] 111. In: McAlpine, J.F., Peterson, B.V., Shewell, G.E., Teskey, H.J., Vockeroth, J.R. & Wood, D.M. (co-ords.) Manual of Nearctic Diptera, Volume 2. Agriculture Canada Monograph 28: 1271–1281.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 409

Marinho, M.A.T., Junqueira, A.C.M., Paulo, D.F., Esposito, M.C., Villet, M.H. & Azeredo-Espin, A.M.L. (2012) Molecular phylogenetics of Oestroidea (Diptera: Calyptratae) with emphasis on Calliphoridae: Insights into the inter-familial relationships and additional evidence for paraphyly among blowflies. Molecular Phylogenetics and Evolution 65: 840–854. Marshall, M.R. (1960) Sternite modification in males of the Drosophilidae (Diptera). Annals of the Entomological Society of America 53: 133–137. McAlpine, D.K. (1985) The Australian genera of Heleomyzidae (Diptera: Schizophora) and a reclassifica- tion of the family into tribes. Records of the Australian Museum 36: 203–251. McAlpine, D.K. (1991) Relationships of the genus Heterocheila (Diptera: Sciomyzoidea) with description of a new family. Tijdschrfit voor Entomologie 134: 193–199. McAlpine, D.K. (1996) Relationships and classification of the Pseudopomyzidae (Diptera: Nerioidea). Proceedings of the Linnean Society of New South Wales 116: 223–232. McAlpine, D.K. (2008) New extant species of Ironic flies (Diptera: Ironomyiidae) with notes on irono- myiid morphology and relationships. Proceedings of the Linnean Society of New South Wales 129: 17–38. McAlpine, J.F. (1981) Morphology and terminology – adults [Chapter] 2. In: McAlpine, J.F., Peterson, B.V., Shewell, G.E., Teskey, H.J., Vockeroth, J.R. & Wood, D.M. (co-ords.) Manual of Nearctic Diptera, Volume 1. Agriculture Canada Monograph 27: 9–63. McAlpine, J.F. (1989) Phylogeny and classification of the Muscomorpha [Chapter] 116. In: McAlpine, J.F. (ed.) Manual of Nearctic Diptera, Volume 3. Agriculture Canada Monograph 32: 1396–1518. Meier, R., Kotrba, M. & Ferrar, P. (1999) Ovoviviparity and viviparity in Diptera. Biological Reviews 74: 199–258. Melander, A.L. (1950) Taxonomic notes on some smaller Bombyliidae (Diptera). The Pan-Pacific Entomologist 26: 139–156. Michelsen, V. (1996) Neodiptera: new insights into the adult morphology and higher level phylogeny of Diptera (Insecta). Zoological Journal of the Linnean Society 117: 71–102. Moulton, J.K. & Wiegmann, B.M. (2007) The phylogenetic relationships of flies in the superfamily Empidoidea (Insecta: Diptera). Molecular Phylogenetics and Evolution 43: 701–713. Nagatomi, A. & Liu, N. (1994) Apystomyiidae, a new family of Asiloidea (Diptera). Acta Zoologica Academiae Scientiarum Hungaricae 40: 203–218. Nagatomi, A., Saigusa, T., Nagatomi, H. & Lyneborg, L. (1991) Apsilocephalidae, a new family of the Orthorrhaphous Brachycera (Insecta, Diptera). Zoological Science 8: 579–591. Nakayama, H. (2012) Complex asymmetric male genitalia of Anevrina Lioy (Diptera: Phoridae). Arthropod Structure and Development 41: 35–49. Neugart, C., Schneeberg, K. & Beutel, R.G. (2009) The morphology of the larval head of Tipulidae (Diptera, Insecta) — the dipteran groundplan and evolutionary trends. Zoologischer Anzeiger 248: 213–235 Neumann, H. (1958) Der Bau und die Funktion der männlichen Genitalapparate von Trichocera annu- lata Meig. und Tipula paludosa Meig. (Dipt. Nematocera). Deutsche Entomologische Zeitschrift N.F. 5: 235–298. Oosterbroek, P. & Courtney, G. (1995) Phylogeny of the nematocerous families of Diptera (Insecta). Zoological Journal of the Linnean Society, 115: 267–311. Oosterbroek, P. & Theowald, Br. (1991) Phylogeny of the Tipuloidea based on characters of larvae and pupae (Diptera, Nematocera). Tijdschrift voor Entomologie 134: 211–267. Ovtshinnikova, O.G. (1986) Musculature of male genitalia in the superfamily Stratiomyioidea (Diptera). Entomological Review 65: 118–132. Ovtshinnikova, O.G. (1989) Musculature of the male genitalia of Brachycera-Orthorrhapha (Diptera). Trudy Zoologicheskogo Instituta Akademiyia Nauk SSR 190: 1–166 [In Russian]. Ovtshinnikova, O.G. (1994a) Musculature of the male genitalia of Pantophthalmidae and systematic position of this family among Brachycera Orthorrhapha (Diptera). Dipterological Research 5: 213–221. Ovtshinnikova, O.G. (1994b) On the homology of male genitalia sclerites of Brachycera Orthorrhapha and Cyclorrhapha (Diptera) based on musculature. Dipterological Research 5: 263–269.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 410 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Ovtshinnikova, O.G. (2003) The taxonomic position of the genusRhaphiomidas Osten Sacken, 1877 (Diptera, Mydidae) in the superfamily Asiloidea, based on the structure of muscles of the male genitalia. Entomological Review 83: 696–701. Ovtshinnikova, O.G. & Yeates, D.K. (1998) Male genital musculature of Therevidae and Scenopinidae (Diptera: Asiloidea): structure, homology and phylogenetic implications. Australian Journal of Entomology 37: 27–33. Palmer, C.M., Ovtshinnikova, O.G. & Yeates, D.K. (2000) Male genitalia of Exeretonevra Macquart (Diptera: Xylophagidae): structure, function and phylogenetic implications. Australian Journal of Entomology 39: 270–274. Pape, T. (1992) Phylogeny of the Tachinidae family-group (Diptera: Calyptratae). Tijdschrift voor Entomologie 135: 43–86. Pape, T. & Arnaud, P.H. (2001) Bezzimyia — a genus of native New World Rhinophoridae (Insecta, Diptera). Zoologica Scripta 30: 257–297. Pape, T., Blagoderov, V. & Mostovski, M.V. (2011) Order DIPTERA Linneaus, 1758. In: Zhang, Z.-Q. (Ed.) Animal biodiversity: an outline of higher-level classification and survey of taxonomic richness: pp. 222–229. Paramonov, N.M. (2004) Taxonomic position of the families Tipulidae and Limoniidae in Nematocera (Diptera) with account of characters of the male genital skeleton and muscles. Entomological Review 84: 15–30. Petersen, F.T., Meier, R., Kutty, S.N. & Wiegmann, B.M. (2007) The phylogeny and evolution of host choice in the Hippoboscoidea (Diptera) as reconstructed using four molecular markers. Molecular Phylogenetics and Evolution 45: 111–122. Peterson, M.J., Bertone, M.A., Wiegmann, B.M. & Courtney, G.W. (2010) Phylogenetic synthesis of morphological and molecular data reveals new insights into the higher-level classification of Tipuloidea (Diptera). Systematic Entomology 35: 526–545. Pollock, J.N. (1973) A comparison of the male genitalia and abdominal segmentation in Gasterophilus and Glossina (Diptera), with notes on the gasterophilid origin of the tsetse flies.Transactions of the Royal Entomological Society of London 125: 107–124. Ribeiro, G.C. (2006) Homology of the gonostylus parts in crane flies, with emphasis on the families Tipulidae and Limoniidae (Diptera, Tipulomorpha). Zootaxa 1110: 47–57. Ribeiro, G.C. (2008) Phylogeny of the Limnophilinae (Limoniidae) and early evolution of the Tipulomorpha (Diptera). Invertebrate Systematics 22: 627–694. Richter, V.A. & Ovtshinnikova, O.G. (1996) On the structure of male and female genitalia in Palaearctic nemestrinids (Diptera, Nemestrinidae). International Journal of Dipterological Research 7: 241–249. Rognes, K. (1997) The Calliphoridae (Blowflies) (Diptera: Oestroidea) are not a monophyletic group. Cladistics 13: 27–66. Rohdendorf, B.B. (1964) The historical development of two-winged insects. Trudy paleontogicheskogo Instituta Akademiyia Nauk SSR 100: 1–311 [In Russian]. Rotheray, G.E. & Gilbert, F. (2008) Phylogenetic relationships and the larval head of the lower Cyclorrhapha (Diptera). Zoological Journal of the Linnean Society 153: 287–323. Schneeberg, K. & Beutel, R.G. (2011) The adult head structures of Tipulomorpha (Diptera, Insecta) and their phylogenetic implications. Acta Zoologica 92: 316–343. Schneeberg, K., Friedrich, F, Courtney, G.W., Wipfler, B. & Beutel, R.G. (2012) The larvae of Nymphomyiidae (Diptera, Insecta) – ancestral and highly derived? Arthropod Structure & Development 41: 293–301. Shatalkin, A.I. (1995) Palaearctic species of Pseudopomyzidae (Diptera). Russian Entomological Journal 3: 129–145. Shatalkin, A.I. (2012) The homologies in structure of male genitalia of Cyclorrhapha (Diptera). Caucasian Entomological Bulletin 8: 321–327, pls. 21-22 [In Russian]. Sinclair, B.J. (1992) Phylogenetic interpretation of the Brachycera (Diptera) based on the larval mandible and associated mouthpart structures. Systematic Entomology 17: 233–252.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 411

Sinclair, B.J. (2000) Morphology and terminology of Diptera male terminalia. [Chapter] 1.2. In: Papp, L. and B. Darvas (eds.) Contributions to a manual of Palaearctic Diptera (with special reference to flies of economic importance). Volume 1. General and Applied Dipterology. Science Herald, Budapest: pp. 53–74. Sinclair, B.J., Borkent, A. & Wood, D.M. (2007) The male genital tract and aedeagal components of the Diptera with a discussion of their phylogenetic significance.Zoological Journal of the Linnean Society 150: 711–742. Sinclair, B.J. & Cumming, J.M. (2006) The morphology, higher-level phylogeny and classification of the Empidoidea (Diptera). Zootaxa 1180: 1–172. Sinclair, B.J., Cumming, J.M. & Wood, D.M. (1994) Homology and phylogenetic implications of male genitalia in Diptera – Lower Brachycera. Entomologica Scandinavica 24: 407–432. Sinclair, B.J. & Kirk-Spriggs, A.H. (2010) Alavesia Waters and Arillo — a Cretaceous-era genus discovered extant on the Brandberg Massif, Namibia (Diptera: Atelestidae). Systematic Entomology 35: 268–276. Snodgrass, R.E. (1935) Principles of insect morphology. McGraw-Hill, New York, NY, 667 pp. Spangenberg, R., Hünefeld, F., Schneeberg, K. & Beutel, R.G. (2012) The male postabdomen and reproductive system of Bibio marci Linnaeus, 1758 (Hexapoda: Diptera: Bibionidae). Journal of Zoological Systematics and Evolutionary Research 50: 264–288. Steyskal, G.C. (1957) The postabdomen of male acalyptrate Diptera.Annals of the Entomological Society of America 50: 66–73. Stuckenberg, B.R. (2001) Pruning the tree: a critical review of classifications of the Homeodactyla (Diptera, Brachycera), with new perspectives and an alternative classification. Studia dipterologica 8: 3–41. Sturtevant, A.H. (1926) The seminal receptacles and accessory glands of the Diptera, with special reference to the Acalypterae. Journal of the New York Entomological Society 34: 1–24. Theodor, O. (1976) On the structure of the spermathecae and aedeagus in the Asilidae and their importance in the systematics of the family. Israel Academy of Sciences and Humanities, Jerusalem, 278 pp. Trautwein, M.D., Wiegmann, B.M. & Yeates, D.K. (2010) A multigene phylogeny of the fly superfamily Asiloidea (Insecta): taxon sampling and additional genes reveal the sister-group to all higher flies (Cyclorrhapha). Molecular Phylogenetics & Evolution 56: 918–930. Trautwein, M.D., Wiegmann, B.M. & Yeates, D.K. (2011) Overcoming the effects of rogue taxa: evolutionary relationships of the bee flies. PLoS Currents Tree of Life. May 5 (last modified: 2012 Apr 23), doi: 10.1371/currents.RRN1233. Ulrich, H. (1972) Zur Anatomie des Empididen-Hypopygiums (Diptera). Veroffentlichungen der Zoologischen Staatssammlung München 16: 1–27. Ulrich, H. (2006) Evidence and error in current interpretations of the brachyceran male genitalia. Sixth International Congress of Dipterology, Abstracts Volume. Fukuoka, Japan: p. 282. Webb, D.W. (1981) Hilarimorphidae. In: McAlpine, J.F., Peterson, B.V., Shewell, G.E., Teskey, H.J., Vockeroth, J.R. & Wood, D.M. (co-ords.) Manual of Nearctic Diptera, Volume 1. Agriculture Canada Monograph 27: 603–605. Wiegmann, B.M., Mitter, C. & Thompson, F.C. (1993) Evolutionary origin of the Cyclorrhapha (Diptera): tests of alternative morphological hypotheses. Cladistics 9: 41–81. Wiegmann, B.M., Trautwein, M.D., Winkler, I.S., Barr, N.W., Kim, J.-W., Lambkin, C., Bertone, M.A., Cassel, B.K., Peterson, K.J., Bayless, K.M., Heimberg, A.M., Wheeler, B.M., Peterson, K.J., Pape, T., Sinclair, B.J., Skevington, J.H., Blagoderov, V., Caravas, J., Kutty, S., Schmidt-Ott, U., Kampmeier, G.E., Thompson, F.C., Grimaldi, D.A., Beckenbach, A.T., Courtney, G.W., Friedrich, M., Meier, R. & Yeates, D.K. (2011) Episodic radiations in the fly tree of life. Proceedings of the National Academy of Sciences 108: 5690–5695. Wiegmann, B.M., Tsaur, S.-C., Webb, D.W., Yeates, D.K. & Cassel, B.K. (2000) Monophyly and relationships of the Tabanomorpha (Diptera: Brachycera) based on 28S ribosomal gene sequences. Annals of the Entomological Society of America 93: 1031–1038. Winteron, S.L. (2004) Are hind coxal knobs a synapomorphy for therevids? An unusual new species of Anabarhynchus Macquart from Australia (Diptera: Therevidae: Therevinae). Zootaxa 413: 1–8.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 412 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

Winterton, S.L. & Irwin, M.E. (2008) Kaurimyia gen. nov.: discovery of Apsilocephalidae (Diptera: Therevoid clade) in New Zealand. Zootaxa 1779: 38–44. Wood, D.M. (1991) Homology and phylogenetic implications of male genitalia in Diptera; the ground plan. In: Weismann, L, Országh, I. & Pont, A.C. (eds.) Proceedings of the Second International Congress of Dipterology. The Hague, Netherlands: pp. 255–284. Wood, D.M. & Borkent, A. (1989) Phylogeny and classification of the Nematocera. In: McAlpine, J.F. (ed.) Manual of Nearctic Diptera. Volume 3. Agriculture Canada Monograph 32: 1333–1370. Woodley, N.E. (1989) Phylogeny and classification of the “Orthorrhaphous” Brachycera [Chapter] 115. In: McAlpine, J.F. (ed.) Manual of Nearctic Diptera. Volume 3. Agriculture Canada Monograph 32: 1371–1395. Woodley, N.E., Borkent, A. & Wheeler, T.A. (2009) Phylogeny of the Diptera. In: Brown, B.V., Borkent, A., Cumming, J.M., Wood, D.M. & Zumbado, M. (eds.) Manual of Central American Diptera. Volume 1. NRC Research Press, Ottawa, ON, pp. 79–94. Yang, L., Wiegmann, B.M., Yeates, D.K. & Irwin, M.E. (2000) Higher-level phylogeny of the Therevidae (Diptera: Insecta) based on 28S ribosomal and elongation factor-1a gene sequences. Molecular Phylogenetics and Evolution 15: 440–451. Yeates, D.K. (1992) Towards a monophyletic Bombyliidae (Diptera): the removal of the Proratinae (Diptera: Scenopinidae). American Museum Novitates No. 3051, 30 pp. Yeates, D.K. (1994) The cladistics and classification of the Bombyliidae (Diptera: Asiloidea).Bulletin of the American Museum of Natural History No. 219, 191 pp. Yeates, D.K. (2002) Relationships of extant lower Brachycera (Diptera): a quantitative synthesis of morphological characters. Zoologica Scripta 31: 105–121. Yeates, D.K. & Irwin, M.E. (1992) Three new species ofHeterotropus Loew (Diptera: Bombyliidae) from South Africa with descriptions of the immature stages and a discussion of the phylogenetic placement of the genus. American Museum Novitates No. 3036, 25 pp. Yeates, D.K. & Irwin, M.E. (1996) Apioceridae (Insecta: Diptera): cladistic reappraisal and biogeography. Zoological Journal of the Linnean Society 116: 247–301. Yeates, D.K., Irwin, M.E. & Wiegmann, B.M. (2003) Ocoidae, a new family of asiloid flies (Diptera: Brachycera: Asiloidea), based on Ocoa chilensis gen. and sp.n. from Chile, South America. Systematic Entomology 28: 417–431. Yeates, D.K. & Wiegmann, B.M. (1999) Congruence and controversy: toward a higher-level phylogeny of Diptera. Annual Review of Entomology 44: 397–428. Yeates, D.K. & Wiegmann, B.M. (2012) The impact of the Manual of Nearctic Diptera on phylogenetic dipterology. In: Festschrift commemorating the coordinators of the Manual of Nearctic Diptera and their contributions to building the Canadian National Collection of Insects. Part 3. The Canadian Entomologist 144: 197–205. Yeates, D.K., Wiegmann, B.M., Courtney, G.W., Meier, R., Lambkin, C. & Pape, T. (2007) Phylogeny and systematics of Diptera: two decades of progress and prospects. Zootaxa 1668: 656–590. Zatwarnicki, T. (1996) A new reconstruction of the origin of ermoneuran hypopygium and its implications for classification (Insecta: Diptera). Genus 7: 103–175. Zloty, J., Sinclair, B.J. & Pritchard, G. (2005) Discovered in our backyards: a new genus and species of a new family from the Rocky Mountains of North America (Diptera, Tabanomorpha). Systematic Entomology 30: 248–266.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 413

Appendix List of characters of Diptera male terminalia. Characters are traced on cladograms (Figs 1–4) and discussed further in the text. 1. Vasa deferentia configuration: (0) extended caudally, not arching back anteriorly; (1) U-shaped, apically approximated. 2. Accessory gland: (0) separate or not continuous with vasa deferentia; (1) continuous with vasa deferentia. 3. Diptera sperm pump: (0) absent; (1) present. 4. Parameres: (0) paired anteroventral apodemes absent; (1) paired anteroventral apodemes present. 5. Aedeagus: (0) separate from parameres; (1) partially enclosed and fused to parameral sheath, encircling the aedeagus. 6. Accessory gland: (0) undivided internally; (1) divided into two or three chambers. 7. Ventral plate: (0) absent; (1) present. 8. Aedeagus: (0) at least partly sclerotized; (1) entirely membranous. 9. Ejaculatory apodeme: (0) present; (1) absent. 10. Ejaculatory apodeme position: (0) anterior to sperm chamber, functioning as piston to compress chamber; (1) ventral to sperm chamber, pushing out spermatophore as it forms. 11. Lateral ejaculatory processes: (0) absent; (1) present. 12. Epandrium and hypandrium: (0) fused ring; (1) separate. 13. Parameres: (0) sperm pump enclosed at base of parameral sheath; (1) sperm pump lying on long, concave parameral sclerite. 14. Ejaculatory apodeme apex: (0) attached to anterior margin of sperm chamber (endophallus); (1) ventrally articulated with phallus; (2) separated with sperm pump from phallus by short to long sperm duct; (3) ejaculatory apodeme greatly reduced, sperm pump lying on concave parameral sclerite; (4) fused and continuous with phallus. 15. Ejaculatory apodeme: (0) simple, without posterior projection; (1) with a posterior extension into the sperm sac (endoaedeagal process). 16. Hypandrium and gonocoxites: (0) separate; (1) completely fused, no seams. 17. Lateral ejaculatory processes: (0) external at apex of ejaculatory apodeme; (1) small, retracted within base of phallus; (2) small, not retracted within base of phallus, closely associated with base of aedeagal tines. 18. Aedeagal tines: (0) absent; (1) present. 19. Gonocoxal apodemes: (0) processes projecting at most little beyond margin of hypandrium; (1) processes elongate, extending well beyond anterior margin of hypandrium; (2) confined to anterior margin, lacking process. 20. Epandrium articulation: (0) fused to or lying flat on hypandrium; (1) articulated on gonocoxites or hypandrium; (2) articulated with prolongation of hypandrium. 21. Gonostylus articulation: (0) transverse; (1) oblique to dorsoventral.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access 414 B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415

22. Aedeagus: (0) separate or partially fused with paramere; (1) completely fused with parameral sheath forming phallus. 23. Gonostyli: (0) present; (1) absent. 24. Epiproct (tergite 10): (0) present; (1) absent. 25. Gonostyli: (0) apical; (1) subapical. 26. Phallus: (0) undivided; (1) subdivided into a dorsal guide and a ventral needle-like component. 27. Epandrium: (0) shallowly notched or posterior margin straight; (1) posterior margin deeply emarginate, forming U-shape with basal connection. 28. Subepandrial sclerite: (0) absent (membranous, at least in part of length); (1) sclerotized along its entire length from base of phallus to hypoproct, forming subepandrial sclerite; (2) reduced, restricted to apex of epandrium. 29. Subepandrial sclerite: (0) undivided laterally; (1) divided laterally into bacilliform sclerites (processus longi), extending to apex of epandrial lamellae. 30. Surstyli: (0) absent; (1) functionally present; (2) articulated with epandrium. 31. Postgonites: (0) absent; (1) present. 32. Phallic plate: (0) absent; (1) present. 33. Hypandrium (fused with gonocoxite): (0) elongate; (1) greatly shortened. 34. Hypopygium: (0) unrotated or not permanently rotated; (1) rotated permanently between 45-90°; (2) rotated and lateroflexed to right, functionally circumverted; (3) circumverted, rotated through 360°. 35. Bacilliform sclerite: (0) articulated with phallic plate or base of phallus; (1) fused to hypandrium. 36. Male terminalia symmetry: (0) symmetrical; (1) asymmetrical around left-side displaced foramen. 37. Phallapodeme: (0) absent; (1) present. 38. Sclerites of segment 7: (0) present; (1) not distinguishable. 39. Sternite 8: (0) separate; (1) united or fused to epandrium. 40. Phallapodeme: (0) ingrowth of the hypandrium; (1) articulated. 41. Position of hypopygium: (0) symmetrical, in medial line of abdomen; (1) asymmetrical, on right side of abdomen. 42. Postgonites: (0) adducted against epandrium; (1) adducted against hypandrium. 43. Circumversion: (0) final rotation of circumversion completed after eclosion; (1) full 360° rotation completed entirely within puparium. 44. Distiphallus: (0) unmodified; (1) flexible, slender, desclerotized and microtrichose; (2) glans present; (3) elongate and coiled; (4) elongate and directly anteriorly. 45. Surstylus vestiture: (0) prensisetae absent; (1) prensisetae present. 46. Phallapodeme: (0) fultelliform sclerite absent; (1) fultelliform sclerite present. 47. Pregonites: (0) absent or short; (1) elongate. 48. Phallic plate: (0) short; (1) elongate. 49. Syntergosternite: (0) multiple sclerites between segment 6 and terminalia, asymmetrical; (1) single sclerite (fusion of sternites 7 and 8), symmetrical. 50. Tergite 6: (0) as large as preceding tergite; (1) reduced and short relative to preceding tergite. Downloaded from Brill.com09/30/2021 12:01:19PM via free access B.J. Sinclair et al. / Insect Systematics & Evolution 44 (2013) 373–415 415

51. Segment 8 sclerites: (0) present; (1) absent. 52. Sternite 7: (0) present; (1) absent. 53. Cerci: (0) present; (1) absent. 54. Sclerites of segment 6-8: (0) separate and asymmetrical; (1) fused, symmetrical and containing 2 pairs of spiracles. 55. Epiphallus: (0) absent; (1) present. 56. Postgonal apodeme: (0) absent; (1) present. 57. Phallus armature (small sclerotized teeth) on distalmost ventral and ventrolateral surfaces: (0) absent; (1) present. 58. Phallus, dorsal wall of distiphallic tube: (0) not modified; (1) sclerotized and forming the dorsal plate. 59. Mesohypophallic sclerotization mid-ventrally along distiphallus: (0) absent or not developed; (1) present.

Downloaded from Brill.com09/30/2021 12:01:19PM via free access