Phylogenetic Analysis of the Subfamily Ototretinae (Coleoptera: Lampyridae)

Phylogenetic analysis of the subfamily Ototretinae (Coleoptera: Lampyridae)

KRISTYNA JANISOVA a & MILADA BOCAKOVA b*

aDepartment of Botany and Zoology, Faculty of Science, Masaryk University, Terezy Novakove 64, CZ-62100 Brno, Czech Republic bDepartment of Biology, Faculty of Education, Palacky University, Purkrabska 2, CZ-77140 Olomouc, Czech Republic

Abstract The first phylogenetic analysis of the subfamily Ototretinae was performed. A data matrix of 42 taxa and 51 morphological characters was assembled, containing all genus - group taxa of the Ototretinae. The monophyly of the Ototretinae was supported. The preferred hypothesis, the majority rule consensus tree generated using implied weights under parsimony criterion showed Baolacus , Falsophaeopterus, Stenocladius to be the most basal splits, followed by divergence of Oculogryphus and Brachypterodrilus . All the other Ototretinae having a circular pit in posterior angles distally formed a crown clade of “higher” Ototretinae proposed here to be the tribe Ototretini. This Ototretini clade was shown to be composed of three lineages: 1. the Drilaster clade, 2. Picodrilus clade, and 3. Lamellipalpus clade. Within Drilaster clade, Emasia was recovered as sister to Drilaster + Flabellototreta clade. North American Brachylampis was shown to originate as sister to Asiatic genus Picodrilus. Within Lamellipalpus clade, Ceylanidrilus and Hydaspoides were recovered among the most basal splits. The genus Lamellipalpodes was shown to be paraphyletic with respect to the Lamellipalpus . Formerly independent subfamily Ototretadrilinae formed a sister group of (Lamellipalpus + Lamellipalpodes ) + Hyperstoma clade. Ceylonese Eugeusis + Harmatelia clade was sister to Gorhamia , and all the three genera were recovered as closely related to ((Lamellipalpus +Lamellipalpodes ) + Hyperstoma ) + Ototretadrilus clade.

Key words Phylogeny, Ototretinae, Lampyridae, morphology

Introduction The Ototretinae belongs to the family Lampyridae, a diverse group of beetles known for their ability of bioluminescence. Lampyridae, or fireflies, include 7 currently recognized subfamilies 1

(Janisova &Bocakova 2013) and almost 2000 species distributed worldwide. While most fireflies use light signals in sexual selection, adults of the subfamily Ototretinae are largely non-luminous, or their light organs are small. The vast majority of ototretine genera are Oriental, with the only exception of the genus Brachylampis Van Dyke, 1939 from California. Most of the ototretines were placed in Drilidae (Wittmer 1944) on the basis of external similarities, namely the shape of pronotum and elytra. Such Drilidae included various largely elateroid genera, namely members of families as Rhagophthalmidae, Phengodidae, Omethidae, Lampyridae, Lycidae. The subfamily Ototretinae was proposed by McDermott (1964), who recognized the extrafamilial synonymy of the Oriental genus Ototreta and Japanese Drilaster (Drilidae). Many other former drilid genera were recognized as ototretine fireflies by Crowson (1972) and this concept was adopted by Lawrence and Newton (1995). Current concept of the subfamily was established by Janisova & Bocakova (2013). The phylogenetic relationships of the Ototretinae and its position within the superfamily Elateroidea have not been clarified yet. Branham &Wenzel (2001, 2003) rejected the monophyly of Ototretinae and proposed Ototretinae split. Consequently, several ototretine genera as Drilaster , Stenocladius , and Harmatelia were excluded from Lampyridae to Elateroidea incertae sedis (Branham 2010). These analyses were exclusively based on external morphological characters and consequently, suffered from parallelisms and convergences. Another recent analyses based on adult morphology (Lawrence et al. 2011) supported paraphyly of Lampyridae (incl. Ototretinae) and Ototretinae (or more exactly Ototretadrilus ) were shown to be basal cantharoid splits (Lawrence et al. 2011). The monophyly of Ototretinae was rejected by a recent unpublished analysis of morphological characters of Lampyridae (Jeng 2008) who presented ototretines as basal paraphyletic grade within Lampyridae. These trees were inconsistent with analyses of large molecular datasets, which have mostly supported the monophyly of both the Lampyridae (incl. Ototretinae) and Ototretinae (Bocakova et al. 2007, Sagegami-Oba 2007, Kundrata et al. 2014). Conversely, analyses of highly variable 16S sequences rejected the monophyly of Lampyridae and Ototretinae (Suzuki 1997, Stanger-Hall 2007). The latter study, although based on 3-gene analyses, was probably affected by the exclusive use of highly variable 16S sequences (18S and cox1 missing) for Stenocladius and Rhagophthalmus (Rhagophthalmidae), and consequently proposed these two genera to be members of Lampyrinae + Luciolinae + Cyphonocerinae clade. While inclusion of the Ototretinae within Lampyridae was supported by analyses of different DNA fragments (see above), the position of the Ototretinae within Lampyridae was challenged. In contrast to morphology based studies, Ototretinae were mostly recovered as a crown clade of Lampyridae (Bocakova et al. 2007, Kundrata et al. 2014).

The Ototretinae are fireflies of short and broad body with head partially covered by pronotum. The eyes are mostly small to medium sized, widely separated. The antennae are filiform to flabellate, extending over pronotum. Labrum is transverse, some taxa present extremely enlarged terminal maxillary and labial palpomeres. Pronotum is transverse with anterior angles rounded, posterior angles triangular, projected obliquely posteriorly, often with a circular pit at base. The biology of the group is poorly known. Both sexes are sometimes fully winged ( Drilaster ), while larviform females were reported for Stenocladius (Ohba et al.1997). Females of many other genera are unknown and are probably brachypterous or apterous. The light production of Ototretinae is weak, or the luminescence of some genera has not been documented yet, although some taxa have abdominal luminous spots as in Harmatelia (McDermott 1965), or terminal abdominal ventrites lighter as in Brachylampis (Van Dyke 1939). In the present study, we take advantage of the recently established taxonomic framework for the subfamily Ototretinae (Janisova & Bocakova 2013; Jeng et al. 2007, 2011) and investigate the phylogeny of Ototretinae based on external morphology and male and female terminalia. To overcome the shortcomings of previous authors, taxon sampling used in this study was substantially expanded and covers all ototretine genera. Based on phylogenetic relationships obtained, the evolution of bioluminescence in adult Lampyridae was tested.

Materials and methods Taxa To test the phylogenetic relationships of Ototretinae, a data matrix of 42 taxa was assembled, consisting of 32 ingroup and 10 outgroup taxa. The ingroup included species representing all Ototretinae genus-group taxa. Preferentially, the type species and one additional species were included to test the monophyly of the genera. For large genera as Drilaster several species were included representing presumed lineages of individual subgenera. The examination of specimens was mainly carried out after the dissection. A specimen of Oculogryphus was unavailable for this study and therefore, its characters were coded from the published descriptions and illustrations (Jeng et al. 2007). For the outgroup, members of other lampyrid subfamilies (Lampyrinae, Luciolinae, Photurinae, Cyphonocerinae) were examined and coded, as well as representatives of several other closely related elateroid families as Lycidae, Cantharidae, and Phengodidae. To eliminate the effect of outgroup choice, multiple outgroup approach was applied.

List of taxa: ELATEROIDEA: outgroups

LYCIDAE Libnetis commendabilis

PHENGODIDAE Phengodes plumosa

CANTHARIDAE Cantharis decipiens

LAMPYRIDAE: Luciolinae Hotaria parvula

LAMPYRIDAE: Photurinae Photuris sp.

LAMPYRIDAE: Cyphonocerinae Cyphonocerus sp.

LAMPYRIDAE: Lampyrinae Lucidina biplagiata Pyrocoelia fumosa Lamprorhiza splendidula Lampyris noctiluca

LAMPYRIDAE: Ototretinae Drilaster axillaris Drilaster subvittatus Drilaster medioniger Drilaster agcoensis Drilaster fuscicollis Flabellototreta fruhstorferi Flabellototreta sp. Emasia dentata Ceylanidrilus bipartitus Hydaspoides kanarensis Ototretadrilus atritarsis Ototretadrilus krombeini Gorhamia compressicornis Eugeusis ramicornis Harmatelia bilinea Lamellipalpodes sp. Lamellipalpus bombayensis Lamellipalpodes annandalei Lamellipalpodes montanus Hyperstoma marginata Hypersoma wittmeri Picodrilus drescheri Picodrilus sp.

Brachylampis sanguinicollis Brachypterodrilus pallidipes Brachypterodrilus atricolor Brachypterodrilus minutus Falsophaeopterus fruhsthorferi Falsophaeopterus jacobsoni Stenocladius davidis Baolacus lajoyei Oculogryphus fulvus

Characters Characters applied here derive largely from external and internal morphology of adult males. Characters of females are omitted in the current analysis, as ototretine females are largely unknown, except for genera Drilaster and Stenocladius . Considering females of Drilaster are fully winged, while those of Stenocladius are larviform, scoring of morphological characters of females would result in a large number of unknown and inapplicable characters states. Similarly, larval characters were mainly omitted as larvae of most ototretine taxa are unknown. Fifty one morphological characters are included in the current analysis (Table 1). Of these, 11 characters refer to external morphology, while the largest suite of characters is provided by male genitalia morphology and the shape of terminal abdominal segments. Missing character states were represented as „?“ and inapplicable characters were coded by a dash (-). Most of the characters (46) were coded into binary states, characters 5, 8, 9, 11, and 25 were coded into multistates. The characters were treated as unordered. The list of characters and their states is presented in Table 1, the data matrix is shown in Table 2.

Table 1. Character list and scoring of states (figures refer to Janisova & Bocakova 2013)

1. Circular pit at base of posterior pronotal angles 0 – absent 1 – present

2. Terminal palpomere of maxillary palpi: 0 – small, at most 3x longer than penultimate one 1 – enlarged and strongly flattened, at least 5x longer than penultimate one

3. Terminal palpomeres of labial palpi: 0 – small, at most 3x longer than palpomere 2 1 – enlarged, at least 8x longer than palpomere 2

4. Bipectinate antennae: 0 – absent 1 – present

5. Head position: 0 – exposed ( Phengodes, Cantharis, Lamellipalpus ) 1 – partly exposed 2 – entirely covered by pronotum ( Cyphonocerus, Lucidina, Pyrocoelia, Lamprorhiza, Lampyris )

6. Mandibles: 0 – small, normal sized 1 – prominent ( Phengodes, Lamellipalpus )

7. Vitreous spots on pronotum 0 – absent 1 – present ( Lampyris, Pyrocoelia, Lamprorhiza )

8. Antennal lobes 0 – short, stout ( Cyphonocerus, Flabellototreta fruhsthorferi, Eugeusis, Harmatelia, Hyperstoma, Brachypterodrilus atricolor, Stenocladius ) 1 – greatly elongate, considerably flattened ( Hydaspoides ) 2 – elongate, rounded in cross-section ( Phengodes ) 3 – absent

9. Antennal insertions 0 – widely separated 1 – moderately approximate 2 – approximate

10. Photic organs on terminal abdominal segments in males: 0 – absent 1 – present

11. Phallobase, emargination of outer margin: 0 –absent 1 – trapezoidal ( D. subvittatus Fig. 69-71, Flabellototreta sp.) 2 – squarish ( Lamprorhiza ) 3 – shallow ( Cyphonocerus )

12. Phallobase, median projection on outer margin: 0 – absent 1 – present ( Emasia – Fig. 87, Ceylanidrilus – Fig. 81, Hydaspoides – Fig. 86, Ototretadrilus – Fig. 107, Picodrilus – Fig. 88, Brachypterodrilus pallidipes – Fig. 118, B. atricolor – Fig. 121, Brachylampis )

13. Phallobase in lateral view: 0 – flattened 1 – hemispherical

14. Parameres, long rod-like projection emerging from ventrobasal portion of each paramere: 0 – absent 1 – present ( Flabellototreta – Fig. 80a)

15. Phallus in ventral view: 0 – straight, lateral margins parallel-sided to tapered 1 – constricted distally, knob-like ( Hydaspoides , Falsophaeopterus )

16. Phallus in lateral view: 0 – straight 1 – deflected

17. Male terminal and penultimate tergum: 0 – separate 1 – fused, jointly trilobed (Figs. 51, 52, 53, 54)

18. Phallus, apical portion: 0 – simple 1 – either with tubular organs, or ventral plate ( Drilaster - Fig. 66-77)

19. Parameres, distal portion: 0 – simple 1 – with two thorns, thorn on inner margin considerably hooked ( Flabellototreta - Fig. 78, 80)

20. Phallobase with a broad shallow emargination, its lateral margins forming long sharp projections:

0 – absent 1 – present (Flabellototreta sp.)

21. Distal portion of phallus: 0 – simple 1 – forming two rod-like projections (Emasia - Fig. 87)

22. Parameres, distal portion:

0 – simple 1 – with minute hooked projection in lateral view (Fig. 82)

23. Parameres, inner margin: 0 – simple 1 – considerably corrugated (Fig. 81)

24. Parameral apex: 0 – simple, rounded 1 – securiform or concave ( Gorhamia, Eugeusis, Harmatelia)

25. Parameres, distal setae: 0 – absent 1 – present, forming a bundle of 1 – 4 setae ( Picodrilus, Brachylampis ) 2 – present, setae scattered on oblique, flattened area ( Gorhamia, Eugeusis, Harmatelia)

26. Phallus in distal portion with lateral plates: 0 – absent 1 – present (Ototretadrilus, Gorhamia, Eugeusis, Harmatelia, Lamellipalpodes, Lamellipalpus, Hyperstoma -Figs. 93a, 99a)

27. Phallus, position of lateral plates: 0 – longitudinal, almost parallel to oblique with parameres ( Ototretadrilus, Lamellipalpus, Lamellipalpodes, Hyperstoma ) 1 – almost orthogonal with parameres ( Gorhamia, Eugeusis, Harmatelia - Figs. 104-106, 110- 115)

28. Phallus, ventral fibrilar organ in distal half 0 – absent 1 – present ( Hyperstoma, Ototretadrilus, Gorhamia, Eugeusis, Harmatelia -Figs. 103, 105, 108, 111)

29. Phallus, sharp ventroapical projection in lateral view: 0 – absent

1 – present ( Ototretadrilus, Lamellipalpus, Lamellipalpodes, Hyperstoma - Figs. 97, 100, 103, 108)

30. Parameres, inner margin with conspicuous sharp projection distally: 0 – absent 1 – present (Ototretadrilus atritaris, Otoretadrilus krombeini, Eugeusis, Lamellipalpodes, Lamellipalpus, Hyperstoma )

31. Parameres basally: 0 – simple 1 – attached to a large mediodorsal sclerite (Lamellipalpus - Fig. 94b, Lamellipalpodes, Hyperstoma)

32. Parameres distally: 0 – parallel-sided, or flattened 1 –divergent, V-shaped, apex broadly hemispherical ( Picodrilus - Fig. 88)

33. Apex of parameres: 0 – rounded, U-shaped in cross-section 1 – flattened, petaloid (Brachypterodrilus pallidipes, B. atricolor, B.minutus., Fig. 116, 119, 122)

34. Apex of parameres, distal hook: 0 – absent 1 – present (Brachypterodrilus pallidipes, B. minutus)

35. Male genitalia in ventral view 0 – slender, elongate, at least 2x as long as wide ( Drilaster subvittatus, Falsophaeopterus, Stenocladius, Baolacus, outgroup) 1 – stout, broad (as long as wide or slightly longer)

36. Phallobase: 0 – simple 1 – H-shaped, outer margin widely elliptically emarginate medially ( Baolacus , Fig. 134)

37. Parameres, inner margin: 0 – simple 1 – with a conspicuous projection in ¾ of paramere length ( Baolacus , Fig. 134)

38. Phallobase: membranous rod-like projection arising from lateral attachments of phallobase

0 – absent 1 – present ( Falsophaeopterus jacobsoni , Fig. 131)

39. Parameres basally: 0 – simple 1 – bifurcate (each with a thin lateral projection) ( Lucidina, Photuris )

40. Base of phallus: 0 – simple 1 – projected proximally to outer margin of phallobase forming two deflexed processes (Hotaria, Cyphonocerus )

41. Parameres: ventral margin with a sharp projection medially 0 – absent 1 – present ( Cyphonocerus )

42. Male terminal abdominal tergum, distal margin: 0 – rounded 1 – emarginate ( Lampyris, Lamprohiza, Pyrocoelia, Lucidina )

43. Phallobase, proximally: 0 – symmetrical 1 – laterally distorted ( Lampyris, Pyrocoelia, Lucidina )

44. Parameres: digitiform attenuation in distal ¼ - 2/5 0 – absent 1 – present ( Lampyris, Pyrocoelia )

45. Phallus, terminal portion subdivided in 2 internal pouches: 0 – no 1 – yes (Pyrocoelia)

46. Phallobase, proximal margin: 0 – rounded 1 – transversely straight, forming a strengthened ridge ( Gorhamia, Eugeusis, Harmatelia - Figs. 104a, 110a, 113a)

47. Base of phallus, two long, proximal, mostly divergent projections:

0 – absent ( Drilaster, Flabellototreta, Emasia , outgroup) 1 – present

48. Larvae, bioluminescence: 0 – absent 1 – present

49. Male terminal sternum with a vitreous spot basally: 0 – absent 1 – present

50. Male genitalia, phallus: 0 – well sclerotized 1 – weakly sclerotized to membranous ( Picodrilus, Drilaster agcoensis )

51. Legs, presence of tarsal lobes: 0 – tarsomeres 1-4 lobed ( Libnetis ) 1 – only tarsomere 4 lobed (Ototretinae, Hotaria, Photuris, Lucidina ) 2 – tarsomeres 3-4 lobed ( Phengodes, Cantharis )

Cladistic analysis The data matrix was assembled and edited in a word processor in nexus format. Phylogenetic analyses were performed using maximum parsimony approach as implemented in TNT (Goloboff et al. 2008). The shortest trees were obtained under New Technology Search (sectorial search parameters: rss and css - globally swap every 2 changes in sectors below 75, and every 10 changes in sector above 75) from different random seeds. After obtaining the shortest trees, implied weighting was applied to reduce the influence of homoplastic characters (Goloboff 1993) with the default concavity constant (k=3.0). Support for individual clades was calculated using standard bootstrapping and symmetric resampling of the original matrix with 1000 replications. Character state reconstructions for individual nodes were displayed on the trees using map synapomorphies option in TNT.

Results and discussion A parsimony analysis of Ototretinae data matrix with equal weights and characters treated as non- additive retrieved 100 most parsimonious trees of 102 steps length (CI = 0.578, RI = 0.803). The majority-rule consensus tree of the shortest trees is shown in Fig. 1. The monophyly of Ototretinae was supported in 80 percent of fundamental shortest cladograms, i.e. deep relationships of ototretine (Baolacus , Falsophaeopterus (incl. Mimophaeopterus), Stenocladius ) were unresolved in the strict

consensus tree. Basal relationships of Ototretinae genera on the majority-rule consensus trees were well resolved. The backbone relationships of Ototretinae were recovered with optimal support (80- 95%, Fig. 2). The majority-rule consensus tree showed subsequent separation of basal ototretine lineages as Baolacus , Falsophaeopterus + Mimophaeopterus clade, Stenocladius , Oculogryphus , and Brachypterodrilus, which is sister to the remaining Ototretinae. The subsequent analysis under implied weights showed better resolution and higher support values. The strict consensus of resulting 24 shortest trees is shown in Fig. 3. The monophyly of Lampyridae and Ototretinae was always recovered (100%), although the bootstrap support and symmetric resampling values were low (Fig. 4). Deep relationships of the Ototretinae were unresolved as the sequence of splitting of Baolacus, Falsophaeopterus, Stenocladius , and other Ototretinae was not clarified. Then, the preferred hypothesis supported separation of three clades: Oculogryphus , Brachypterodrilus , and the “higher” Ototretinae. The latter lineage designated here as the tribe Ototretini was recovered in all trees derived from morphological characters and is considered to be well supported by sharing a synapomorphic feature, the presence of a circular pit near apex of posterior angles of pronotum (Janisova & Bocakova 2013). As the clade is supported by a single character, the support values from bootstrapping and symmetric resampling were low, although there is no inferred reversal, i.e. the character in question shows no homoplasy. This Ototretini clade was formed by three major lineages: 1. Drilaster clade = ( Drilaster + Flabellototreta ) + Emasia , 2. Picodrilus clade = Picodrilus + Brachylampis, and 3. Lamellipalpus clade = Hydaspoides + Ceylanidrilus + (((Lamellipalpus + Lamellipalpodes ) + Hyperstoma ), + Ototretadrilus ) + ((Eugeusis + Harmatelia ) + Gorhamia )). Our morphological analyses supported Emasia to be a sister group of morphologically close genera Drilaster and Flabellototreta . Within Picodrilus clade, the American genus Brachylampis was recovered as sister to Asiatic Picodrilus . Finally, the Lamellipalpus clade is composed of a large number of genera, several of which are known to share presence of large terminal maxillary and labial palpomeres. Although sometimes this character was considered to be a synapomorphy of Lamellipalpus , Lamellipalpodes , Hyperstoma , and Eugeusis , our analysis rejected monophyly of this clade and therefore, we propose large terminal palpomeres evolved twice in Ototretini, 1x in Eugeusis and 1x in Lamellipalpus + Lamellipalpodes + Hyperstoma clade. The comparison of results of current analysis of morphological data is difficult, as the taxon sampling of published DNA sequence analyses is fragmentary. The molecular phylogenetic analysis of the Elateriformia (Bocakova et al. 2007) supported the monophyly of the Ototretinae, although the internal relationship of the group was slightly different from the current morphological trees. In agreement to presented analysis of morphological data, molecular analyses supported that Falsophaeopterus (sg. Mimophaeopterus ) diverged early from the core Ototretinae clade, designated here as the tribe

Ototretini. Sister group relationships of Drilaster and Flabellototreta proposed here by morphological data, were rejected by molecular data (Bocakova et al. 2007), although the set of Drilaster species included in morphological and molecular analyses were not identical. Conversely, analysis of DNA sequences supported Flabellototreta as sister to Emasia , and Drilaster should be sister to (Ototretadrilus + (Flabellototreta + Emasia )) clade. Using the inferred phylogenetic trees, the evolution of bioluminescence in adult Lampyridae was tested, although inferred phylogenetic relationships showed deep polytomies and therefore, were unable to resolve the question whether the ancestor of Lampyridae was luminescent in adult stage.

Conclusions This analysis is the first study of the relationships of all genus-group taxa of the Ototretinae. The main conclusions are:

1. The monophyly of the Ototretinae from all over the world is supported as the American Brachylampis was shown to be sister to Asiatic Picodrilus . 2. Deep relationships of Ototretinae lineage show subsequent divergence of five clades. The first five splits are Baolacus , Falsophaeopterus , Stenocladius , Oculogryphus , and Brachypterodrilus . The latter is sister to the “higher” Otrotretinae clade, i.e. the tribe Ototretini. 3. Within Ototretini three main lineages were recovered: the Drilaster clade, Picodrilus clade, and Lamellipalpus clade. 4. Monophyly of Ototretini taxa with large terminal palpomeres was rejected and Lamellipalpodes was recovered as paraphyletic. American Brachylampis evolved within Asiatic Picodrilini clade.

Acknowledgements We would like to thank to the following entomologist and institutions for providing material in their charge: A. Taghavian (MNHN), M. Barclay (BMNH), D. Kavanaugh (CASC), M. Brancucci (NHMB), and Z. Simmons (OUMN). The type material in MNHN and BMNH was examined under EU-funded Synthesys (Synthesis of Systematic Resources) projects (FR-TAF-353, GB-TAF-1982). This study has been supported by grants IGA_PdF_2015029 from Palacky University Olomouc (Czech Republic) and CZ.1.07/2.3.00/20.0166 (MB) from European Social Fund and the Ministry of Education of the Czech Republic.

References Bocakova, M., Bocak, L., Hunt, T. Teraväinen, M., Vogler, A. P. 2007. Molecular phylogenetics of Elateriformia (Coleoptera): evolution of bioluminiscence and neoteny. Cladistics 23 (2007): 477–496.

Branham, M. A., Wenzel, J. W., 2001. The evolution of bioluminescence in cantharoids (Coleoptera: Elateroidea). Fla. Entomol. 84: 565-586.

Branham, M. A., Wenzel, J. W., 2003. The origin of photic behavior and the evolution of sexual communication if fireflies (Coleoptera: Lampyridae). Cladistics 84: 565-586.

Branham, M. (2010) Lampyridae Latreille, 1817, pp. 141-149. In : Leschen R. A., Beutel R. G., and J. F. Lawrence (Eds) Handbook of Zoology. Part 38, Vol. 2. Morphology and Systematics (Elateroidea, Bostrichiformia, Cucujoidea). Walter de Gruyter, Berlin, New York, 786 pp.

Crowson, R. A. (1972) A review of the classification of Cantharoidea (Coleoptera), with the definition of two new families, Cneoglossidae and Omethidae. Revista de la Universidad de Madrid , 21 (82), 35–77.

Goloboff, P. A. 1993. Estimating character weights during tree search. Cladistics , 9(1): 83–91.

Goloboff, P. A., Farris, J. S. and K. Nixon. 2008. TNT: a free program for phylogenetic analysis. Cladistics , 24: 774–786.

Janisova, K. & Bocakova, M. (2013): Revision of the subfamily Ototretinae (Coleoptera: Lampyridae), Zoologischer Anzeiger , 252 (1), 1–19.

Jeng, M.L., M. Engel, and P.S. Yang. 2007a. Oculogryphus , a remarkable new genus of fireflies from Asia (Coleoptera: Lampyridae). American Museum Novitates Nr. 3600: 1–19.

Jeng, M.L. 2008. Comprehensive phylogenetics, systematice, and evolution of neoteny of Lampyridae (Insecta: Coleoptera). Ph.D. Thesis. Graduate Faculty of the University of Kansas, 388 pp.

Jeng, M.L., Branham M., M. Engel 2011. A second species of Oculogryphus Coleoptera,Lampyridae), with notes on the phylogenetic affinities of the genus. ZooKeys 97: 31–38 (2011), doi: 10.3897/zookeys.97.1223

Kundrata, R., Bocakova, M. & Bocak, L. (2014): The comprehensive phylogeny of the superfamily Elateroidea (Coleoptera: Elateriformia), Molecular Phylogenetics and Evolution, 76, 162-171.

Lawrence, J. F. & Newton, A. F. (1995). Families and subfamilies of Coleoptera (with selected genera, notes, references and data on family-group names). In J. Lawrence, J. F. Pakaluk & S. A. Ślipi ński (Eds) Biology, phylogeny, and classification of Coleoptera (pp. 779-1092). Museum i Instytut Zoologii PAN, Warzawa.

Lawrence, J. F., Slipinski, A., Seago, A. E., Thayer, M. K., Newton, A. F. & Marvaldi, A. E. (2011). Phylogeny of the Coleoptera Based on Morphological Characters of Adults and Larvae. Annales Zoologici , 61, 1-217.

McDermott, F. A. 1964. The taxonomy of the Lampyridae (Coleoptera). Trans. Amer. Entomol. Soc. 90: 1–72.

McDermott, F. A. 1965. The Pterotinae (Coleoptera: Lampyridae). Ent. News. April pp. 98-104.

Ohba, N., Y. Goto and I. Kawashima 1997. Behavior and adult female morphology of firefly, genus Stenocladius (Coleoptera: Lampyridae) in Japan. Scientific Report of the Yokosuka City Museum 45: 23–37 (in Japanese with English summary).

Sagegami-Oba, R., Takahashi, N., Oba, Y. 2007. The evolutionary process of bioluminescence and aposematism in cantharoid beetles (Coleoptera: Elateroidea) inferred by the analysis of 18S ribosomal DNA. Gene 400 (1-2): 104-113 .

Stanger-Hall, K. F., Lloyd, J.E., Hillis, D. M. 2007. Phylogeny of North American fireflies (Coleoptera: Lampyridae): Implications for the evolution of light signals. Mol. Phy. Evol. 45(1): 33-49.

Suzuki, H. 1997. Molecular phylogenetic studies of Japanese fireflies and their mating systems (Coleoptera: Cantharoidea). Tokyo Metropolitan Univ. Bull. Nat. Hist. 3: 1-53.

Van Dyke, E.C. 1939. New species and subspecies of west American Coleoptera. Pan-Pac. Entomol. Wittmer, W. 1944. Catalogue des Drilidae E. Oliv. (Coleoptera-Malacodermata). Rev. Soc. Entomol. Arg. 12: 203–221.

Table 2. The data matrix of 51 characters and 42 taxa of Ototretinae

Libnetis_commendabilis 0000100320000000?000000000-00000000000000?000000000 Phengodes_plumosa 00010102100000110000000000-00000000000000-000001002 Cantharis_decipiens 0000000310001000?000000000-00000-0-000000?010-?0002 Hotaria_parvula 00001003210000010000000000-000000000000100000001101 Photuris_sp. 00001003210000000000000000-000000000001000000001101 Cyphonocerus_sp. 00012000213000010000000000-000000000000110000001101 Lucidina_biplagiata 00002003200000000000000000-000000000001001100001101 Pyrocoelia_fumosa 00002013210000000000000000-000000000000001111001101 Lamprorhiza_splendidula 00002013212010010000000000-000000000000001000001101 Lampyris_noctiluca 00002013210000010000000000-000000000000001110001101 Drilaster_axillaris 10001003100000010100000000-000000010000000000001101 Drilaster_subvittatus 10001003101000010100000000-000000000000000000001101 Drilaster_medioniger 10001003100000010100000000-000000010000000000001101 Drilaster_agcoensis 10001003100000010100000000-000000010000000000001111 Drilaster_fuscicolli 10001003100000010100000000-000000010000000000001101 Flabellototreta_fruhstorferi 10001000200001010010000000-000000010000000000001101 Flabellototreta_sp. 10001003201001010011000000-000000010000000000001101 Emasia_dentata 10001003200100010000100000-000000010000000000001101 Ceylanidrilus_bipartitus 10001003100110010000011000-000000010000000000011101 Hydaspoides_kanarensis 10001001100110110000000000-000000010000000000011101 Ototretadrilus_atritarsis 100010031001100100000000010111000010000000000011101 Ototretadrilus_krombeini 100010032001100100000000010111000010000000000011101 Gorhamia_compressicornis 100010031000100000000001211100000010000000000111101 Eugeusis_ramicornis 111010001000100000000001211100000010000000000111101 Harmatelia_bilinea 100010001000100000000001211100000010000000000111101 Lamellipalpodes_sp. 111010031000100010000000010011100010000000000011101 Lamellipalpus_bombayensis 111001030000100010000000010011100010000000000011101 Lamellipalpodes_annandalei 111010031000100010000000010011100010000000000011101 Lamellipalpodes_montanus 111010031000100110000000010011100010000000000011101 Hyperstoma_marginata 111010001000000110000000010111100010000000000011101 Hypersoma_wittmeri 111010001000000110000000010111100010000000000011101 Picodrilus_drescheri 10001003100100010000000010-000010010000000000011111 Picodrilus_sp. 10001003100100010000000010-000010010000000000011111 Brachylampis_sanguinicollis 10001003000100010000000010-000000010000000000011101 Brachypterodrilus_pallidipes 00001003200100010000000000-000001110000000000011101 Brachypterodrilus_atricolor 00001000200100010000000000-000001010000000000011101 Brachypterodrilus_minutus 0?00100?200000010000000000-000001110000000000011101 Falsophaeopterus_fruhstoferi 00001003200000100000000000-000000000000000000011101 Falsophaeopterus_jacobsoni 00001003200000100000000000-000000000010000000011101 Stenocladius_davidis 00001000200000010000000000-000000000000000000011101 Baolacus_lajoyei 00001003200000000000000000-000000001100000000011101 Oculogryphus_fulvus 00000003200100010000000000-000000000000100010011?01