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Rhagophthalmidae are a small beetle family known from the eastern Palaearctic and Oriental realms. Rhagophthalmidae are closely related to railroad worms (Phengodidae) and fireflies (Lampyridae) with which they share highly modified paedomorphic females and the ability to emit light. Currently, Rhagophthalmidae include 66 species classified in the following 12 genera: Bicladodrilus Pic, 1921 (two spp.), Bicladum Pic, 1921 (two spp.), Dioptoma Pascoe, 1860 (two spp.), Diplocladon Gorham, 1883 (two spp.), Dodecatoma Westwood, 1849 (eight spp.), Falsophrixothrix Pic, 1937 (six spp.), Haplocladon Gorham, 1883 (two spp.), Menghuoius Kawashima, 2000 (three spp.), Mimoochotyra Pic, 1937 (one sp.), Monodrilus Pic, 1921 (two spp. in two subgenera), Pseudothilmanus Pic, 1918 (two spp.), and Rhagophthalmus Motschulsky, 1854 (34 spp.). The replacement name Haplocladon gorhami Kundrata, nom. nov. is proposed for Diplocladon hasseltii Gorham, 1883b (described in subgenus Haplocladon) which is preoccupied by Diplocladon hasseltii Gorham, 1883a. The genus Reductodrilus Pic, 1943 is tentatively placed in Lampyridae: Ototretinae. Lectotypes are designated for Pseudothilmanus alatus Pic, 1918 and P. marginalis Pic, 1918. Interestingly, in the eastern part of their distribution, Rhagophthalmidae have remained within the boundaries of the Sunda Shelf and the Philippines demarcated by the Wallace Line, which separates the Oriental and Australasian realms. This study is intended to be a first step towards a comprehensive revision of the group on both genus and species levels. Additionally, critical problems and prospects for rhagophthalmid research are briefly discussed.

Bioluminescence, the emission of visible light in a living organism, is an intriguing phenomenon observed in different species and environments. In terrestrial organisms, the bioluminescence is observed mainly in beetles of the Elateroidea superfamily (Coleoptera). Several phylogenetic studies have been used different strategies to propose a scenario for the origin and evolution of the bioluminescence within this group, however some of them showed incongruences, mainly about the relationship of the bioluminescent families. In order to increase the number of molecular markers available for Elateroidea species and to propose a more accurate phylogeny, with high supported topology, we employed the Next-Generation Sequencing (NGS) methodology to perform the RNA-Seq analysis of luminescent (Elateridae, Phengodidae, Rhagophthalmidae, and Lampyridae) and non-luminescent (Cantharidae) species of Neotropical beetles. We used the RNA-Seq data to construct a calibrated phylogeny of Elateroidea superfamily using a large number of nuclear molecular markers. The results indicate Lampyridae and Phengodidae/Rhagophthalmidae as sister-groups, suggesting that the bioluminescence evolved later in Elateridae than other families (Lampyridae, Phengodidae, and Rhagophthalmidae), and indicating the Upper Cretaceous as the period for the main diversification of Elateroidea bioluminescent species.

Simple Summary In the era of phylogenomics, new molecular sequencing and computational techniques can aid in resolving phylogenetic relationships that were previously intractable by morphological or limited molecular data. In this study, we used anchored hybrid enrichment—designed to recover DNA sequences from hundreds of single-copy orthologous genes—to resolve the phylogeny of the Elateridae (click-beetles) and establish their placement within superfamily Elateroidea. The resulting data were compatible with published transcriptomes, allowing for integrating our dataset with previously published data. Using a wide range of analyses on these molecular data, we tested hypotheses long-debated in the morphological literature and also the robustness of our phylogenetic inferences. Our results placed the bioluminescent lampyroids (fireflies and relatives) within the click-beetles, challenging the current classification of Elateridae, Lampyridae, Phengodidae, and Rhagophthalmidae. However, despite the large amount of molecular data analyzed, a few nodes with conflicting phylogenetic signals could not be unambiguously resolved. Overall, we recovered well-resolved tree topologies that will serve as a framework for further systematic and evolutionary studies of click-beetles. This work further demonstrates that the click-beetle lineage contains not only pest wireworms, but also many species that benefit agriculture. Abstract Click-beetles (Coleoptera: Elateridae) are an abundant, diverse, and economically important beetle family that includes bioluminescent species. To date, molecular phylogenies have sampled relatively few taxa and genes, incompletely resolving subfamily level relationships. We present a novel probe set for anchored hybrid enrichment of 2260 single-copy orthologous genes in Elateroidea. Using these probes, we undertook the largest phylogenomic study of Elateroidea to date (99 Elateroidea, including 86 Elateridae, plus 5 non-elateroid outgroups). We sequenced specimens from 88 taxa to test the monophyly of families, subfamilies and tribes. Maximum likelihood and coalescent phylogenetic analyses produced well-resolved topologies. Notably, the included non-elaterid bioluminescent families (Lampyridae + Phengodidae + Rhagophthalmidae) form a clade within the otherwise monophyletic Elateridae, and Sinopyrophoridae may not warrant recognition as a family. All analyses recovered the elaterid subfamilies Elaterinae, Agrypninae, Cardiophorinae, Negastriinae, Pityobiinae, and Tetralobinae as monophyletic. Our results were conflicting on whether the hypnoidines are sister to Dendrometrinae or Cardiophorinae + Negastriinae. Moreover, we show that fossils with the eucnemid-type frons and elongate cylindrical shape may belong to Eucnemidae, Elateridae: Thylacosterninae, ancestral hard-bodied cantharoids or related extinct groups. Proposed taxonomic changes include recognition of Plastocerini as a tribe in Dendrometrinae and Hypnoidinae stat. nov. as a subfamily within Elateridae.

Bioluminescent beetles of the superfamily Elateroidea (fireflies, fire beetles, glow-worms) are the most speciose group of terrestrial light-producing animals. The evolution of bioluminescence in elateroids is associated with unusual morphological modifications, such as soft-bodiedness and neoteny, but the fragmentary nature of the fossil record discloses little about the origin of these adaptations. We report the discovery of a new bioluminescent elateroid beetle family from the mid-Cretaceous of northern Myanmar (ca99 Ma), Cretophengodidae fam. nov.Cretophengodes azarigen. et sp. nov. belongs to the bioluminescent lampyroid clade, and would appear to represent a transitional fossil linking the soft-bodied Phengodidae + Rhagophthalmidae clade and hard-bodied elateroids. The fossil male possesses a light organ on the abdomen which presumably served a defensive function, documenting a Cretaceous radiation of bioluminescent beetles coinciding with the diversification of major insectivore groups such as frogs and stem-group birds. The discovery adds a key branch to the elateroid tree of life and sheds light on the evolution of soft-bodiedness and the historical biogeography of elateroid beetles.

Pseudothilmanus Pic, 1918 is a genus of beetles currently placed in Rhagophthalmidae. The species P. alatus Pic, 1918 and P. marginatus Pic, 1918 were described based on single holotypes from India with little descriptive information. Later, in a revision of the genus, a new specimen of P. alatus was examined, but also without specific locality. This paper provides the description of the wing venation and new distributional records for this genus, including the first record in Nepal.

Bioluminescence in beetles is found in the Elateroidea superfamily, which includes Elateridae, Lampyridae, Rhagophthalmidae, and Phengodidae families. Despite sharing the same biochemical system, it is unclear whether the lanterns and the bioluminescent system share a common origin or not. To better understand the molecular differentiation of photogenic from non-photogenic tissues and the molecular physiology of these tissues, we compared the transcriptional profiles of the lateral lanterns and fat body in Phrixothrix hirtus railroad worm (Phengodidae) and the abdomen of the non-luminescent Chauliognathus flavipes (Cantharidae). We observed gene products involved in pheromone biosynthesis in all tissues and four distinct opsin gene products in C. flavipes, more than expected when compared to the transcriptional analysis of lampyrid eyes. We also observed several gene products potentially associated with bioluminescence in P. hirtus lantern and in C. flavipes body, whereas in the fat body of P. hirtus we found only a single copy of luciferase homolog enzyme. Altogether these results suggest that the photogenic tissue of phengodid lanterns has distinct origin from fireflies lanterns, and also the existence of a latent bioluminescent potential in the fat body tissue of some Elateroidea families.

The pH-insensitive beetle luciferases cloned from Rhagophthalmidae, Phengodidae, and Elateridae exhibit great potential application as reporter assays for monitoring gene expression. At present, however, only one luciferase has been reported from the enigmatic and predominantly Asian distributed luminous family Rhagophthalmidae. Here, we cloned the second rhagophthalmid luciferase from the Chinese glow-worm Menghuoius giganteus (Rhagophthalmidae: Elateroidea) by combining reverse transcription polymerase chain reaction (RT-PCR) with rapid amplification of complementary DNA ends (RACE). The luciferase consisted of 546 amino acids and showed high identity to that of Rhagophthalmus ohbai (90.4%). The recombinant M. giganteus luciferase was produced in vitro and exhibited significant bioluminescent activity under neutral conditions (pH 7.8), with low KM for D-luciferin (2.2 μm) and ATP (53 μm). Activity was highest at 10°C and inactivation occurred at 45°C. This luciferase showed pH-insensitivity and maximum emission spectrum at 560 nm. Phylogenetic analyses based on the deduced amino acids indicated a close relationship between the M. giganteus luciferase and that of R. ohbai. These results increase our understanding of rhagophthalmid luciferases and provide a new resource for the application of luciferases.

The collection of Sri Lankan Cantharoidea in the National Museums Colombo was initiated and identified by European and British collectors in the early 1800s. Their collection has been reserved as "Sri Lankan firefly collection" in the National Museums, Colombo. Subsequently no systematic studies have been undertaken in an attempt to confirm the identifications and update the classification of these specimens using recent taxonomic information. During January to June in 2010, we examined 1,214 dry mounted specimens using a stereo microscope to determine external morphology in an attempt to reclassify specimens using up to date taxonomy. The collection originally was arranged into 4 families classified under 27 genera and 63 species. In our opinion, there are 4 families, containing 29 genera and 61 species including 27 Lampyridae species. The collection is clearly marked with labeled specimens indicating any authority for the suggested changes.

Fireflies are one of the best-known examples of luminescent organisms. The limited geographic distribution and rarity of some firefly genera have hindered molecular phylogenetic analysis, resulting in uncertainty in regard to firefly phylogeny. Here, using genome skimming next-generation sequencing, we sequenced 23 Asian firefly species from 15 genera (Lampyridae: 14; Rhagophthalmidae: one) and assembled their mitochondrial genomes (mitogenomes) and nuclear ribosomal DNA (rDNA) repeat unit. The mitogenomes (including 15 mitochondrial genes: COX1-3, ATP6&8, ND1-6&4L, CYTB, 12S, and 16S) were recovered for almost all 23 species; furthermore, three regions of the nuclear rDNA repeat unit (18S, 28S, and 5.8S) were recovered for 22 out of the 23 species. The mitogenomes of 11 genera and 22 species as well as the complete rDNA from 22 species are reported here for the first time. Combined with previously published sequences of mitochondrial and rDNA coding regions, 166 species (170 populations with four overlapping in Lampyridae) were included in the current analyses. We selected different species groups and coding regions to infer phylogenies, and then employed tree certainty (TC) and internode certainty (IC) to quantify any phylogenetic incongruence. Phylogenetic analysis of 18 coding regions (15 mitochondrial genes and three regions of the nuclear rDNA repeat unit) from different species groups showed that the 144-species selection group (excluding 22 species outside Lampyridae) had relatively high TC (101.39). Further phylogenetic analysis of the 144 species using different coding regions indicated that the phylogeny of the 13 coding regions (10 mitochondrial genes: COX1-2, ATP6&8, ND1, ND4-5, CYTB, 12S and 16S; three rDNA regions: 18S, 5.8S, and 28S) demonstrated higher TC (103.02) than the phylogenies based on the 18 coding regions (TC = 101.39), conserved-regions (c-regions, i.e., 12S, 16S, COX1, 18S, and 28S) (TC = 95.11), or conserved-sites (c-sites, TC = 92.31) for the mitochondrial genes. In contrast, the c-sites strengthened the deeper nodes of the 144-species phylogeny compared to the c-regions. All of the 144-species phylogenies using different coding regions (except the c-regions) consistently recovered the monophyly of each of the three luminous families and their combination (Lampyridae, Rhagophthalmidae, and Phengodidae) with high IC support. Our phylogenetic analyses clarified the position of firefly genera Lamprigera, Vesta, Stenocladius, Pyrocoelia, Diaphanes, Abscondita, Pygoluciola, Emeia, Pristolycus, and Menghuoius. We also inferred the evolutionary pattern of adult bioluminescence in Lampyridae based on the phylogenies of 166 and 144 species. Our data suggest that the common ancestor of Lampyridae possessed adult bioluminescence, with a higher loss rate than gain rate of bioluminescence during its lineage evolution. Our results provide insight into Asian firefly phylogeny, and also enrich mitogenome and rDNA data resources for further study.

The evolution of bioluminescence has prompted scientific attention to illuminate phylogenetic relationships of luminescent beetles. However, genomic resources are virtually lacking in rhagophthalmids (Rhagophthalmidae) and their related firefly beetles lampyrids (Lampyridae). Here, we employed the Illumina Hiseq 2000 platform and sequenced the whole-body transcriptomes of the four luminescent beetles: one rhagophthalmid (Rhagophthalmus sp.) and three fireflies (Asymmetricata circumdata, Aquatica ficta, and Pyrocoelia pectoralis). We obtained 55.4, 43.4, 38.6, and 36.7 million clean reads for the four species, respectively. All reads were assembled into contigs from which unigenes were derived. All unigenes were annotated by publicly available databases, and a total of 4325 orthologous genes were identified. Using multiple phylogenetic approaches, our transcriptome data confirmed the distinctiveness of Rhagophthalmidae from Lampyridae, which was also supported by our mitogenome analysis using three newly determined mitogenome sequences and 12 previously published ones. Together, this study is the first report of whole transcriptome sequencing data in Rhagophthalmidae and Lampyridae species, representing a valuable genomic resource for studying the origin and evolution of some remarkable traits in these beetles such as bioluminescence. Moreover, our transcriptome and mitogenome data provide useful phylogenetic information that could be of importance in future studies of phylogenetic inference.

Rhagophthalmus ohbai Wittmer (Iriomote botaru, in Japanese) can be found in the soil environment on several islands in Okinawa Prefecture, Japan. The females of this species have been reported to remain in very close proximity to their eggs for about 2 months while producing a pungent odor. We hypothesized that females produced this odor in order to protect their eggs and/or themselves from pathogens and/or predators. Analysis of this volatile substance by solid-phase microextraction and gas chromatography mass spectrometry (GC–MS) revealed that the odor was attributed to 1,4-naphthoquinone (NQ). We also examined resistance to NQ in four fungi isolated from dead fireflies [Aspergillus tubingensis (Schober) Mosseray 21KI501, A. fumigatus Fresenius 21KI502, Fusarium proliferatum (Matsushima) Nirenberg 21KI503, and Trichoderma asperellum Samuels et al. 21KI504], as well as four common soil bacteria [Escherichia coli (Migula) Castellani and Chalmers type B, Staphylococcus aureus subsp. aureus Rosenbach H209P, Bacillus subtilis (Ehrenberg) Cohn ATCC6633, and Rhodococcus jostii Takeuchi et al. RHA1]. The findings revealed that NQ exhibited antimicrobial activity in all of the test organisms, suggesting that female fireflies release NQ to protect their eggs and newly hatched larvae from pathogens in the soil. We also analyzed the volatiles produced by male fireflies using GC–MS and found that some male fireflies emitted NQ. It is possible that male fireflies produce NQ to protect themselves from pathogenic microorganisms in the soil environments that they inhabit during the day. Although several defensive chemicals produced by fireflies have been identified to date, the production of 1,4-naphthoquinone for chemical defense by R. ohbai has not yet been reported.

Diversity in Peru: 2 subfamilies, 7 genera, 12 species. Recognition: Phengodidae are commonly called glow-worms because the larviform females and larvae have bioluminescent organs in transverse bands along the body. They glow yellow-green using eleven paired photic organs of 2 thoracic through 9th abdominal segments; some genera have head organs that glow red. Some adult males are also bioluminescent, and at least one species has bioluminescent eggs. The adult male body length is 3.3–25 mm; larviform females are generally larger than males. The antenna has 10–12 antennomeres in adult males, and may be filiform, serrate, bipectinate or plumose. Adult males have truncate or dehiscent elytra. Phengodid larvae are slender, mostly well-sclerotized, and rounded in crosssection (compared to the flattened larvae of Lampyridae and Lycidae), resembling those of click beetles. Habitat: These may be nocturnal, active at nights. Winged males may be attracted at light traps. Females may be collected from loose bark while larvae are usually found under leaf litter or rotting logs. The larviform females and larvae are specialist predators of millipedes and other litter arthropods. Notes: The American family Phengodidae is now considered as separate from Asiatic Rhagophthalmidae, although sister group relationships of the families were supported by both morphological evidence (Lawrence et al., 1995), and comprehensive molecular phylogenies (Bocakova et al., 2007; Kundrata et al., 2014). Previous studies suggested rhagophthalmids as a clade of fireflies (Crowson, 1955; Suzuki, 1997). The following checklist is derived from Pic in Titschack (1954; as Drilidae), Zaragoza-Caballero (1984), and specimens housed in SEMC. The family was monographed by Wittmer (1976), but a number of new genera have been added since. Lawrence et al. (2000) reported the minute Penicillophorus (2–4 mm) as occurring in Colombia and Peru. The “railroad worm”, Phrixothrix, occurs in Peru but it is unclear if it is a pest (see Costa et al., 1999). An interactive key to the world fauna is available online (Lawrence et al., 2000); see Zaragoza-Caballero (2010) for an updated key to the genera of the large subfamily Mastinocerinae. Taximastinocerus is reported as new genus record for Peru based on 19 specimens (probably multiple species) collected by Chaboo in 2010–2011 with flight intercept traps and Malaise traps at the CICRA Biological Field Station, Madre de Dios, Peru.

The purpose of this application, under Article 23.9.3 of the Code, is to conserve the usage of the genus-group name Dodecatoma Westwood, 1849 for a group of beetles (family phengodidae or rhagophthalmidae) by suppressing the unused senior homonym Dodecatoma Dufour, 1841, which was proposed for a group of stoneflies (family perlidae).

Kundrata, R. & Bocak, L. (2011). The phylogeny and limits of Elateridae (Insecta, Coleoptera): is there a common tendency of click beetles to soft-bodiedness and neoteny? —Zoologica Scripta, 40, 364–378. Phylogenetic relationships in Elateroidea were investigated using partial 18S and 28S rDNA and rrnl and cox1 mtDNA sequences with special interest in the phylogeny of Elateridae and the position of soft-bodied lineages Drilidae and Omalisidae that had been classified as families in the cantharoid lineage of Elateroidea until recently. Females in these groups are neotenic and almost completely larviform (Drilidae) or brachypterous (Omalisidae). The newly sequenced individuals of Elateridae, Drilidae, Omalisidae and Eucnemidae were merged with previously published datasets and analysed matrices include either 155 taxa with the complete representation of fragments or 210 taxa when some fragments were missing. The main feature of inferred phylogenetic trees was the monophyly of Phengodidae + Rhagophthalmidae + Omalisidae + Elateridae + Drilidae with Omalisidae regularly occupying a basal node in the group; Drilidae were embedded as a terminal lineage in the elaterid subfamily Agrypninae and soft-bodied Cebrioninae were a part of Elaterinae. The soft-bodied males and incompletely metamorphosed females originated at least three times within the wider Elateridae clade. Their atypical morphology has been considered as a result of long evolutionary history and they were given an inappropriately high rank in the previous classifications. The frequent origins of these modifications seem to be connected with modifications of the hormonal regulation of the metamorphosis. The superficial similarity with other soft-bodied lineages, such as Cantharidae, Lycidae, Lampyridae, Phengodidae and Rhagophthalmidae is supposed to be a result of homoplasious modifications of the ancestral elateroid morphology due to the incomplete metamorphosis. The results of phylogenetic analyses are translated in the formal taxonomic classification. Most Drilidae are placed in Elateridae as a tribe Drilini in Agrypninae, whilst Pseudeuanoma and Euanoma are transferred from Drilidae to Omalisidae. The subfamily Cebrioninae is placed in Elaterinae as tribes Cebrionini and Aplastini. Oxynopterini, Pityobiini and Semiotini are lowered from the subfamily rank to tribes and classified in Denticollinae.

Rhagophthalmus fugongensis Li & Liang and Rhagophthalmus lufengensis Li & Ohba are introduced as new to science. The first record of R. tonkineus in China is provided. The male genitalia of R. gibbosulus and R. giganteus are described and illustrated. The female and larva of R. giganteus and the female of R. semisulcatus are illustrated. The distribution of Rhagophthalmus is discussed and a rectification of the relative arrangement of different parts of the male genitalia is provided (the base-piece is on the ventral side of the male genitalia, and the parameres on the dorsal side).

“Starworm” is the common name for larvae and larviform females of the rhagophthalmid genus Diplocladon (Haneda, 1950; 1985). Such a name results from their three longitudinal rows of lights, one dorso-central and two laterals, produced by three spot-like light organs located on contiguous body segments (Harvey, 1952; Braham and Wenzel, 2001; Ohba, 2004). Although Diplocladon was erected by Gorham in 1883, only two species (D. hasseltii Gorham, 1883, and D. haplocladon indicum Gorham, 1903) have been reported from India and Indonesia (Wittmer, 1944). Diplocladon hasseltii is known from both sexes (Haneda, 1950), but D. haplocladon indicum Gorham known only from the male. In addition, Harvey (1952) thought that luminous larvae or larviform females of a few unidentified species of Diplocladon (some up to 5 cm long) lived in Hangzhou (Zhejiang Province, China, Waterhouse, 1889) and Malaya (Gahan, 1924). Ridley (1934) also dealt with some Diplocladon larva from Singapore and pointed out the large (also circa 5 cm long) luminous insects of Clark (1865) from India belonged in this genus. Because Diplocladon males are nonluminous, they are less commonly collected and correctly associated with the conspecific females. Females and larvae are usually edaphic and relatively cryptic, making them also difficult to collect. Perhaps, the scarcity of specimens has made the systematics of Diplocladon problematic. For instance, Diplocadon has been placed in the Drilidae (Gorham, 1883; Olivier, 1910; Wittmer, 1944; Harvey, 1952), Phengodidae (Crowson, 1972), or Rhagophthalmidae (Lawrence et al., 2000; Branham, 2000). Herein, we follow Lawrence et al. (2000). Having been woken up by his barking dog at midnight, Mr. Guojian Liao, a coal worker at the Tongda Coal Company, found a luminous object on a slope about 50 meters from his home [Baiguhe Village, Tongda Township, Huaping County, Lijiang Prefecture, Yunnan Province, China (N26.75541o, E101.10284o; 2234 m)] on June 6, 2006. The object turned out to be a gigantic (circa 9 cm long) luminous worm. At first sight, its body shape and coloration are very similar to those of larvae and larviform females of the luminous beetle genus Rhagophthalmus (Rhagophthalmidae). Rhagophthalmus larvae are predatory but larviform females rarely feed (Ohba, 1997). The luminous worm was kept in a bottle

Phylogenetic relationships in the coleopteran Series Elateriformia (click beetles, jewel beetles, fireflies and allies) were investigated using > 3800 nucleotides of partial nuclear (small and large subunit rRNA genes) and mitochondrial (large subunit rRNA and cytochrome oxidase subunit I) gene sequences. The Elateriformia includes several soft-bodied lineages, some of which retain larviform features in the adult stage (neoteny), and several major bioluminescent groups, including the families Lampyridae (fireflies), Phengodidae and Rhagophthalmidae whose relationships have been contentious. All recognized superfamilies (Elateroidea, Cantharoidea, Byrrhoidea, Buprestoidea, Dascilloidea, Scirtoidea) and 28 of the 37 families, represented in 112 individuals, were included in the analysis. Sequence alignment was based on static and dynamic homology assignments and partial removal of sequences of uncertain homology. Alignment variable regions caused a great deal of uncertainty but also contributed much of the phylogenetic signal that was insufficient to resolve deep relationships when these were removed. The main features of most analyses were the monophyly of Elateroidea + Cantharoidea (= Elateroidea sensu lato), with Omethidae + Telegeusidae frequently occupying the basal node in this group; the affinities of Dascilloidea, Buprestoidea and a (broadly paraphyletic) Byrrhoidea, with unclear relationships among them; and the monophyly of Scirtoidea (including Decliniidae) as a rather distant outgroup to all others. When mapped on the resulting trees, soft-bodied lineages were polyphyletic, contradicting the former Cantharoidea that had been united by this trait. Transitions to neoteny were either simultaneous with, or subsequent to, the origin of soft-bodiedness in a minimum of seven lineages. The bioluminescent groups Lampyridae (including the enigmatic genus Drilaster) and the tightly allied Phengodidae + Rhagophthalmidae were never monophyletic. The former showed close relationship to the species-rich, soft-bodied families Lycidae and Cantharidae, while the latter grouped with poorly resolved lineages at the base of Elateridae (click beetles). Hence, although key features as soft-bodiedness, neoteny and bioluminescence in Coleoptera are largely confined to the Elateriformia, they appear to result from multiple origins, showing the propensity of closely related lineages to acquire similar features independently. © The Willi Hennig Society 2007.

The eyes of male and female Rhagophthalmus ohbai are of very different sizes and possess approximately 3000 and 35 facets, respectively. In the male eye one can distinguish a smaller dorsal region with 500 facets and a larger ventral one with ca. 1800. Ultrastructural differences between them have been described earlier in this journal (Lau and Meyer-Rochow, 2006). Electrophysiological recordings from the two eye areas have now revealed that the ventral region is maximally sensitive to light of 600 nm wavelength, while the dorsal eye region responds maximally to light of 540–560 nm wavelengths. In the dorsal eye region sensitivity to UV-radiation at around 360 nm wavelength, being twice as high as that of the ventral eye region, amounted to ca. one quarter of peak wavelength sensitivity. The regional differences in spectral sensitivity seem to be a reflection of the different tasks of the two eye regions: looking downward to see the yellow light emitted by a female, sensitivity towards longer wavelengths would be advantageous, but looking upward into the twilight sky, sensitivity to shorter wavelength would be a more appropriate adaptation.

Mitochondrial genome organization in the Elateroidea superfamily (Coleoptera), which include the main families of bioluminescent beetles, has been poorly studied and lacking information about Phengodidae family. We sequenced the mitochondrial genomes of Neotropical Lampyridae (Bicellonycha lividipennis), Phengodidae (Brasilocerus sp.2 and Phrixothrix hirtus) and Elateridae (Pyrearinus termitilluminans, Hapsodrilus ignifer and Teslasena femoralis). All species had a typical insect mitochondrial genome except for the following: in the elaterid T. femoralis genome there is a non-coding region between NADH2 and tRNA-Trp; in the phengodids Brasilocerus sp.2 and P. hirtus genomes we did not find the tRNA-Ile and tRNA-Gln. The P. hirtus genome showed a ~1.6kb non-coding region, the rearrangement of tRNA-Tyr, a new tRNA-Leu copy, and several regions with higher AT contents. Phylogenetics analysis using Bayesian and ML models indicated that the Phengodidae+Rhagophthalmidae are closely related to Lampyridae family, and included Drilus flavescens (Drilidae) as an internal clade within Elateridae. This is the first report that compares the mitochondrial genomes organization of the three main families of bioluminescent Elateroidea, including the first Neotropical Lampyridae and Phengodidae. The losses of tRNAs, and translocation and duplication events found in Phengodidae mt genomes, mainly in P. hirtus, may indicate different evolutionary rates in these mitochondrial genomes. The mitophylogenomics analysis indicates the monophyly of the three bioluminescent families and a closer relationship between Lampyridae and Phengodidae/Rhagophthalmidae, in contrast with previous molecular analysis.

The eyes of the winged males and larvi-form, wingless females of the firefly Rhagophthalmus ohbai differ from each other in several respects. Compared with the eyes of the males, those of the females contain fewer (35 versus ca. 3500) and smaller (20 μm versus 24-31 μm) facets and anatomically they are of the apposition type. Their main function appears to be to detect light intensity changes from day to nighttime; resolving power of the female eye must be poor and e-vector discrimination would be absent. The eyes of the males consist of a smaller, dorsal region of ca. 500 om-matidia of about 250 μm length and a larger, ventral region of ca. 2000 ommatidia of about 640 urn length. The microvilli of the dorsal eye region are somewhat wider than those of the ventral region (55 nm versus 45 nm) and are less regularly arranged. A tapetal reflecting layer is only present in the dorsal eye region. The small clear-zone between dioptric apparatus and retina in the dorsal eye region would not allow as good a superposition image to be produced as in the ventral eye region with its 5 times wider clear-zone. The regular orientations of the microvilli in the rhabdoms and the lack of a proper tapetum in the ventral eye region suggest that e-vector discrimination should be possible.

Elateriformia consists of Dascilloidea, Buprestoidea (jewel beetles), Byrrhoidea and Elateroidea (click beetles, fireflies and relatives). Numerous elateroid lineages contain taxa with modified metamorphosis resulting in sexual maturity while retaining larval characters. Additionally, they evolved unique defensive strategies including clicking mechanism, aposematic coloration and bioluminescence. To investigate the phylogenetic position of Elateroidea within Coleoptera, we merged 1048 newly produced 18S rRNA, 28S rRNA, rrnL mtDNA, and cox1 mtDNA sequences for ∼300 elateriform taxa with data from GenBank. The 975-taxa dataset aligned in BlastAlign was analyzed under maximum likelihood criterion. The results agreed in most aspects with the current morphology-based classification and results of molecular studies. Elateriformia were monophyletic and Elateroidea were sister to Byrrhoidea. Further, we analyzed all-data (513 elateriform taxa) and pruned matrix (417 elateriform taxa, all fragments present) using parsimony and maximum likelihood methods to reveal the phylogenetic relationships among elateroid lineages and examine the evolution of soft-bodiedness, neoteny and bioluminescence. We confirmed the monophyly of Elateroidea sensu lato and most of the families, with Telegeusidae inferred in most trees within paraphyletic Omethidae. The clade Artematopodidae+Telegeusidae+Omethidae was a sister to remaining elateroids. All topologies reject the relationships of hard-bodied Elateridae, Eucnemidae, Throscidae and Cerophytidae, formerly supposed to be a monophylum. Eucnemidae and Throscidae formed independent lineages and the position of Cerophytidae was variable - either a sister to Throscidae, or an independent lineage. The Lampyridae+Cantharidae clade was in most trees sister to Phengodidae+Rhagophthalmidae+Omalisidae+Elateridae. Molecular phylogeny of Elateroidea confirmed the multiple origins of soft-bodied, neotenic and light emiting lineages. On the basis of our molecular phylogeny, we place former Telegeusidae as a subfamily in Omethidae.

Rhagophthalmus giallolateralus Ho sp. nov. (Fig. 4 B, 5 D, 5 E, 5 F, 6 C, 6 D, 7 C, 7 D, 8 C, 8 D, 9 A, 10 C, 10 D, 13) Type. Holotype, TAIWAN: Dongjyu, Lienchiang County, Taiwan, 1 male (bathed samples), 26 -IV- 2010, Jian-Hua Wang. (ESRI); Paratype: TAIWAN: Dongjyu, Lienchiang County, Taiwan, 2 males, 2 females, 23 -V- 2011, Hua-Te Fang. (ESRI). Holotype is deposited in ARI; four paratypes are deposited in NMNS and ESRI. Etymology. This species is named for its yellowish margin on the elytra. Giallo- means the color yellow and - lateralus means the lateral margin. Diagnosis. The enlarged flagellar segments 4���9 are similar to R. flavus which occurs in Malaysia and Thailand, but in R. flavus a lens-like sensillum adheres to each enlarged segment (Kawashima and Sat��, 2001). In Matzu Archipelago, R. beigansis and R. giallolateralus can be simply distinguished from males in the elytra, and the genitalia. The elytra of R. giallolateralus have a yellow margin which is lacking in R. beigansis. Also, there are clear differences in the number of segments of the antennae and maxillary palps, and in the relative position and shape of the female labial palps. In R. beigansis the antennae are 5 -segmented, the maxillary palps are 4 segmented, and the labial palps (Fig. 9 A, 9 B) are slender and protrude beyond the anterior margin of the head. In R. giallolateralus the antennae are 8 -segmented, the maxillary palps are 5 segmented, and the labial palps are wider and shorter, almost not reaching the anterior head margin. Male. Body mostly dark brown, dorsal surface covered with light yellow pubescence. Head and pronotum shiny. Head capsule black; compound eyes blackish, extending to upper and lower area, concave dorsally; antennae brown; mandibles dark brown; maxillae dark brown; labium dark brown to brownish; pronotum dark brown; elytra dark brown, with obvious orange-yellowish or yellowish brown on each margin; ventral surface of thorax orangeyellowish; coxae and trochanters orange-yellow, femora mainly brown to dark brown, orange-yellow at base and apex, tibiae and tarsi dark brown; abdomen dark brown to blackish, with light yellow or white markings along posterior margin of segments. Body ovoid from above, blunt at front and thinnest behind, punctuation separated in dorsal view. Head wider than long, semi-circular, widest at basal margin, but narrower than the apical and basal width of pronotum, punctures separated and surrounding antenna and mouthparts; compound eyes meniscus-like in dorsallateral view, concave basally. Antennae 12 -segmented, 1.77 mm in length; scape short and thick; pedicel, similar to scape in shape; 1 st to 3 rd flagellar segments filiform, weakly broad posteriorly with length longer than scape and pedicel; 4 th to 9 th flagellar segments slightly serrate, length and shape are almost the same, lens-like sensillum located at antero-ventral side of 9 th flagellomere; 10 th flagellomere (terminal segment) spindle shaped, slender, and tapering at its apex. Pronotum transverse, semi-circular in dorsal view, anterior margin very broadly rounded with no distinct anterolateral corners and merging into the divergent lateral margins; right angles formed at the junction of lateral and basal margin, basal margin straight or weakly wavy, the widest part of pronotum, but a little narrower than humeral width of elytra; punctation separated, spread uniformly over surface; PW/HW 1.14, PW/PL 1.51, PW/PA 1.17, PW/EW 0.68, PW/EHW 0.83. Elytra elongate, contiguous along inner margin, outer margins slightly expanded; extending to, or slightly before abdominal apex; lateral margin straight and nearly parallel-sided, width contracting over apical 1 / 7; punctation separated, pubescence uniform over surface; EL/PL 5.37, EL/EW 2.40, EW/EHW 1.24. Legs slender; femur with lateral margin straight and parallel-sided; tibia slightly conical, thicker at apex than base; tarsi 5 -segmented, each segment clavate, with 2 claws apically, empodium or arolium not distinguished; HFL/HTL 0.96. Male genitalia 0.9 mm in length, trilobed without punctation and pubescence, basal plate large, covering the basal part of aedeagus and parameres in ventral view, basal margin arcuate and protruded towards base without angle, basal 1 / 2 of lateral margins dispersed, gradually separated towards apex, then subparalleled on apical 1 / 2; aedeagus shorter than parameres, bluntly cone-like, lateral margin arcuate, gradually converging towards bluntly rounded apex, grooves not apparent over surface; parameres dipper-like, apical parts protruding towards apex ventrally and surrounding the outer side of the aedeagus; basal margin narrow, forming an incisive arcuate-shape at the basal end, the outer lateral margin straight, converging rapidly towards inner side near apex, forming right angles at marginal junction, inner side straight in dorsal view, dark bands located at basal 1 / 2. Female. Larviform, 18.52 �� 1.64 mm (range: 16.32���20.44, n= 9) in length, 2.35 �� 0.17 mm (range: 2.18���2.72, n= 9) wide at basal margin of pronotum. Body yellowish or brownish. Head small, with small compound eyes. Antennae 8 -segmented, maxillary palp 5 -segmented, labial palp 3 -segmented. Labial palps are tightly shrunk. There are two sets of luminous organs. The first set is a large luminous organ on 7 th ventral abdominal segment and the second consists of three spot-like luminous organs on most segments. Two on both body sides extending from the mesothorax to the 9 th abdominal segment, and one dorsally from the middle of the mesothorax to the 8 th abdominal segment. Measurement in mm. BL: 10.99 (holotype) (range: 10.84���11.02); HW: 2.35 (range: 2.33���2.38); PL: 1.77 (range: 1.77���1.82); PA: 2.29 (range: 2.29���2.31); PB: 2.68 (2.68���2.99); PW: 2.68 (range: 2.68���2.74); EL: 9.51 (range: 9.51���10.17); EW: 3.97 (range: 3.91���3.97); EHW: 3.21 (range: 3.21���3.30); HFL: 2.12 (range: 2.11���2.15); HTL: 2.20 (range: 2.12���2.20). Distribution. TAIWAN: Dongjyu, Matzu Archipelago, Lienchiang County. Remark. The vegetation in habitats of R. giallolateralus consists of thick grass mainly, or of forests or lofty herbs, e.g. Miscanthus floridulus (Labill.). Adults occur in February to April, females are active between 6: 30 and 8:00 at night. The behavioral display, fecundity and characteristics of the eggs are similar to Rhagophthalmus beigansis. Discussion Rhagophthalmidae species have little capacity to migrate due to the wingless form of adult females, and it would be expected that isolated populations like the two described here would diverge significantly. Two Rhagophthalmus species addressed here were found in Beigan islet and Dongjyu islet separately. These islets are less than 40 km apart, but no population of these species was found in other neighboring islands. With virtually no capacity to migrate the species are totally dependent on the existing habitat of these islets and conservation of these fragile environments is thus very important. Habitat fragmentation and human activities have already been identified as the major stress to such island populations (Atkinson, 1989; Hess, 1990). Since the populations of these two Rhagophthalmus species were distributed in a somewhat fragmentary nature across the islands, further study could evaluate the habitat fragmentation. Wittmer and Ohba (1994) discovered R. ohbai in Iriomote island. Because of its distinctive distribution, ecological habitat, and biological features, the Ministry of the Environment, Japan designated R. ohbai as an emergency conservative species. The species was then investigated in a series of related studies (Ohba, 1997). The islets of Beijan and Dongjyu, are only 6.44 and 2.64 km 2 in area. Because of the instability and vulnerability in island ecology, and the dependence of Rhagophthalmus on the existing habitat, further investigation of the two Rhagophthalmus distributed in Matzu Archipelago is necessary with the aim of protecting the fragile environment. Few investigations thus far have focused on the behavior, habitat and breeding methods of Rhagophthalmidae (Ohba et al., 1996; Ohba, 1997, 2004). Male genitalia have to date presented the most useful characteristics for identification of Rhagophthalmus species (Wittmer & Ohba, 1994; Kawashima & Sat��, 2001; Kawashima & Sugaya, 2003; Li et al., 2008). Males, being nocturnal and non-luminous are difficult to collect (Li et al., 2008). Increased awareness of the genus and its distribution and improved collection methods should hopefully address this situation., Published as part of Ho, Jen-Zon, Chen, Young-Fa, Cheng, Su-Han, Tsai, Xi-Lian & Yang, Ping-Shin, 2012, Two new species of Rhagophthalmus Motschulsky (Coleoptera: Rhagophthalmidae) from Matzu Archipelago, Taiwan with biological commentary, pp. 1-13 in Zootaxa 3274 on pages 9-13, DOI: 10.5281/zenodo.214798, {"references":["Kawashima, I. & Sato, M. (2001) Three new species of the genus Rhagophthalmus (Coleoptera, Rhagophthalmidae) from southeast Asia. Elytra, 29 (2), 423 - 434.","Atkinson, A. (1989) Introduced animals and extinction. In: Western, D. & Pearl, M. (Eds.), Conservation for the Twenty-first Century. Oxford University Press, New York, pp. 54 - 69.","Hess, A. L. (1990) Overview: Sustainable Development and Environmental Management of Small Islands. In: Beller, W., D'Ayala, P. & Hein, P. (Eds.), Sustainable Development and Environmental Management of Small Islands. Paris, France, pp. 3 - 14.","Wittmer, W. & Ohba, N. (1994) Neue Rhagophthalmidae (Coleoptera) aus China und Benachbarten Landern. Japanese journal of entomology, 62, 341 - 350. (in Germany)","Ohba, N. (1997) Breeding of the firefly, Rhagophthalmus ohbai (Coleoptera: Rhagophthalmidae). Science Report of the Yokosuka City Museum, 45, 51 - 55.","Ohba, N., Goto, Y. & Kawashima, I. (1996) External morphology and behavior of Rhagophthalmus ohbai WITTMER, 1994 (Coleoptera; Rhagophthalmidae) and its habitat. Science Report of the Yokosuka City Museum, 44, 1 - 19.","Ohba, N. (2004) Flash communication systems of Japanese fireflies. Integrative and Comparative Biology, 44 (3), 225 - 233.","Kawashima, I. & Sugaya, H. (2003) An additional new species of the genus Rhagophthalmus (Coleoptera, Rhagophthalmidae) from Taiwan, with a key to males of the Taiwanese and Japanese species. Elytra, 31 (2), 353 - 359.","Li, X. Y., Ohba, N. & Liang, X. C. (2008) Two new species of Rhagophthalmus Motschulsky (Coleoptera: Rhagophthalmidae) from Yunnan, south-western China, with notes on known species. Entomological Science, 11, 259 - 267."]}

Rhagophthalmus beigansis Ho sp. nov. (Figs 4 A, 5 A, 5 B, 5 C, 6 A, 6 B, 7 A, 7 B, 8 A, 8 B, 9 B, 10 A, 10 B, 11, 12, 13) Type. Holotype. Male. TAIWAN: Beigan, Lienchiang County, Taiwan, 29 -IV- 2011, Jing-Han Hu. (ARI). Paratypes: TAIWAN: Beigan, Lienchiang County, Taiwan, 4 males, 4 females, 8 -V- 2011, Hua-Te Fang. (ESRI, NMNS). Etymology. This species is named after Beigan islet where it was collected. Diagnosis. This new species is similar to R. ohbai but differs in the male genitalia, the female luminous organ and the appendage of the female head. In the male genitalia of R. beigansis, the posterior basal piece is sharp and a space exists between the paramere and medial lobe; whereas in R. ohbai, the posterior basal piece is cylindrical and the paramere is almost attached to the medial lobe. The female in R. beigansis has three spot-like luminous organs, two on both sides of the body between the mesothorax and the 9 th abdominal segment and one in the middle (dorsally) between them mesothorax and the 8 th abdominal segments. This is significantly different from R. ohbai where the 9 th abdominal segment has no luminous organs (Ohba et al., 1996). In labial palps of R. beigansis also protrudes more at the position of the base attachment than in R. ohbai. Finally the number segments of the antennal and maxillary palps in R. beigansis are 5 and 4, but in R. ohbai, are 7 or 8 and 5. Male. Body mostly dark brown or black, moderately shiny, pubescence light yellow, covering almost all the dorsal surface. Head capsule black, not glossy; compound eyes blackish; antennae brown to orange-yellowish; mandibles dark brown; maxillae dark brown; labrum brownish; pronotum dark brown, darker in center of disk and paler brown towards the sides; elytra dark brown, paler brown towards the base; ventral thorax orange-yellowish or yellowish brown; coxae, trochanters, and femora of all legs orange-yellowish or yellowish brown; tibiae and tarsi brownish or dark brown; abdomen dark brown in anterior segments, then darker brown to blackish in the terminal segments, with orange-yellowish or yellowish brown posterior margins. Body spindle or oval-shaped when viewed from above, tiny punctures scattered on the dorsal side of elytra. Head longer than wide, width of posterior margin wider than the apical margin of pronotum, but slightly narrower than the basal width of pronotum, odontoid protrusions surrounding whole surface of head besides appendages, more clearly and more closely clustered around compound eyes. Antennae 12 segmented, 1.68 mm (range: 1.37���1.68) long; scape cylindrical; barrel-shaped pedicel longer than scape in length; 1 st flagellar segment longer than remaining segments, slightly clavate, broader towards the apex,; 2 nd to 4 th flagellar segments also slightly expanded towards their apices with flagellar segment 2 about half as long as 1, 3 and 4 subequal slightly shorter than 2, 5��� 8 more broadly expanded at their apices than preceding segments, approximately subequal in length and shorter than more basal segments; flagellar segment 9 weakly expanded to one side, (asymmetrically clavate) with a lens-like sensillum located at the antero-ventral side of the segment; 10 th flagellar segment very narrow in apical half. Pronotum approximately semi-circular from above, basal margin slightly less than the width across elytral humeri; apical margin protruding forwardly, broadly rounded and anterolateral corners are broadly rounded and obtuse; basal margin straight or slightly arcuate; posterolateral corners acute and pointed; separated odontoid protrudings spread all over the surface of pronotum, more densely aggregated over central disc, gradually reducing in number from center to edges; PW/HW 1.12, PW/PL 1.46, PW/PA 1.39, PW/EW 0.73, PW/EHW 0.95. Elytra elongate slightly oval in outline, inner margins contiguous along their length and outer margins very slightly convex sided; contracting in apical 1 / 3 where the elytra are narrowest, discrete punctures and pubescence over surface of elytra; EL/PL 4.96, EL/EW 2.48, EW/EHW 1.30. Legs slender, femur slightly enlarged at the middle; tibia straight, slightly thinner than femur; tarsus 5 -segmented, pretarsus with 2 claws and no empodium or arolium obvious between the claws; HFL/HTL 0.95. Male genitalia 0.95 mm long, trilobate, glabrous, with no punctures, odontoid protrusions and pubescence; basal plate large and well sclerotized, covering basal 1 / 2 of parameres when viewed from below, anterior margin blunt and round, slightly arcuate, lateral margins subparallel, with a broad straight vertical groove formed at the center of the apex; aedeagus subequal to parameres in length, tapering to a rounded apex; parameres with strongly developed apices inturning but not reaching to the aedeagus; clear dark banks at basal 1 / 3. Basal plate of male genitalia large and roughly straight in each margin, is a critical characteristic on identify. Female. Larviform, 17.26 �� 2.97 mm (range: 10.48���23.41, n= 27) in length, 2.05 �� 0.30 mm (range: 1.44���2.97, n= 27) wide across basal margin of pronotum. Body color yellowish or brownish. Head small, with small compound eyes. Antennae 5 segmented, maxillary palp 4 segmented, labial palp 3 segmented. Labial palps are protruding beyond the anterior head margin. There are two sets of luminous organs in female. The first set is a large luminous organ on 7 th ventral abdominal segment and the second has three spot-like luminous organs on most segments. Each segment has three small spot-like luminous organs, two on both body sides from mesothorax to 9 th abdominal segments and one on dorsal middle from mesothorax to 8 th abdominal segments. Measurement in mm. BL: 8.86 (holotype)(range: 8.34���11.72); HW: 2.11 (range: 2.06���2.13); PL: 1.62 (range: 1.57���1.64); PA: 1.70 (range: 1.69���1.70); PB: 2.37 (2.37 ��� 2.37); PW: 2.37 (range: 1.2���2.69); EL: 8.04 (range: 8.00��� 8.11); EW: 3.24 (range: 3.20���3.25); EHW: 2.49 (range: 2.41���2.49); HFL: 1.81 (range: 1.79���1.86); HTL: 1.91 (range: 1.88���1.93). Distribution. TAIWAN: Beigan, Matzu Archipelago, Lienchiang County. Remarks. Individuals of R. beigansis are nocturnal throughout their life cycle. The vegetation within the habitat is either thick grass or forest. Larvae prey on millipedes observed in the laboratory. Adults appeared in April to May, with the female displaying the luminous behavior between 7:00 to 8: 30 p. m. (Fig. 10). After mating, the female laid 101.4 �� 26.26 eggs (range: 68���151, n= 10) that were 0.8���1.1mm in size, oval-shaped and yellow-white in color. In addition, eggs were attended (Fig. 11)., Published as part of Ho, Jen-Zon, Chen, Young-Fa, Cheng, Su-Han, Tsai, Xi-Lian & Yang, Ping-Shin, 2012, Two new species of Rhagophthalmus Motschulsky (Coleoptera: Rhagophthalmidae) from Matzu Archipelago, Taiwan with biological commentary, pp. 1-13 in Zootaxa 3274 on pages 4-9, DOI: 10.5281/zenodo.214798, {"references":["Ohba, N., Goto, Y. & Kawashima, I. (1996) External morphology and behavior of Rhagophthalmus ohbai WITTMER, 1994 (Coleoptera; Rhagophthalmidae) and its habitat. Science Report of the Yokosuka City Museum, 44, 1 - 19."]}

Two new species of Rhagophthalmus Motschulsky, R. beigansis Ho sp. nov. and R. giallolateralus Ho sp. nov. collected in a vulnerable environment in two small islets of Matzu Archipelago, Lienchiang County are described and illustrated. This brings the total number of species in this genus to 35, and the number known from small off shore islets and islands to 10. Their importance to the conservation of such small and fragile island habitats is discussed.

Representatives of the beetle family Lampyridae ("fireflies", "lightningbugs") are well known for their use of light signals for species recognition during mate search. However, not all species in this family use light for mate attraction, but use chemical signals instead. The lampyrids have a worldwide distribution with more than 2000 described species, but very little is known about their phylogenetic relationships. Within North America, some lampyrids use pheromones as the major mating signal whereas others use visual signals such as extended glows or short light flashes. Here, we use a phylogenetic approach to illuminate the relationships of North American lampyrids and the evolution of their mating signals. Specifically, to establish the first phylogeny of all North American lampyrid genera, we sequenced nuclear (18S) and mitochondrial (16S and COI) genes to investigate the phylogenetic relationships of 26 species from 16 North American (NA) genera and one species from the genus Pterotus that was removed recently from the Lampyridae. To test the monophyly of the NA firefly fauna we sequenced the same genes from three European lampyrids and three Asian lampyrids, and included all available Genbank data (27 additional Asian lampyrids and a former lampyrid from Asia, Rhagophthalmus). Our results show that the North American lampyrids are not monophyletic. Different subgroups are closely related to species from Europe, Asia and tropical America, respectively. The present classification of fireflies into subfamilies and tribes is not, for the most part, supported by our phylogenetic analysis. Two former lampyrid genera, Pterotus and Rhagophthalmus, which have recently been removed from this family, are in fact nested within the Lampyridae. Further, we found that the use of light as a sexual signal may have originated one or four times among lampyrids, followed by nine or four losses, respectively. Short flashes originated at least twice and possibly three times independently among our study taxa. The use of short flashes as a mating signal was replaced at least once by the use of long glows, and light signals as mating signals were lost at least three times in our study group and replaced by pheromones as the main signal mode.

We present the first cladistic analysis of genera in the family Lampyridae and other closely related beetles. A monophyletic concept of Lampyridae is established. The phylogenetic positions of the luminous cantharoid families [Omalisidae, Rhagophthalmidae and Phengodidae] in relation to Lampyridae are discussed, as well as the implications of the evolution of bioluminescence and photic signaling in this group of beetles. The Rhagophthalmidae appears to include Dioptoma and Diplocladon (formerly located in Phengodidae) and the Phengodidae apparently includes Stenocladius (formerly of Lampyridae). Harmatelia, Drilaster and Pterotus are transferred to Elateroidea insertae sedis and not included in Lampyridae where they were sometimes placed.

Pseudothilmanus Pic, 1918 is redescribed and two species, P. alatus Pic, 1918 and P. marginatus Pic, 1918, are placed in the genus. The subgenus Drilothilmanus Pic, 1918 is synonymized with the nominotypical genus on the basis of low morphological divergence and redundancy in classification. Pseudothilmanus is newly classified in the family Rhagophthalmidae, in contrast to the original placement in Drilidae. Diagnostic characters for Pseudothilmanus and both species placed within the genus are illustrated.