A comprehensive molecular phylogeny of tiger beetles (Coleoptera,Carabidae, Cicindelinae)

Tiger beetles are a remarkable group that captivates amateur entomologists, taxonomists and evolutionary biologists alike. This diverse clade of beetles comprises about 2300 currently described species found across the globe. Despite the charisma and scientific interest of this lineage, remarkably few studies have examined its phylogenetic relationships with large taxon sampling. Prior phylogenetic studies have focused on relationships within cicindeline tribes or genera, and none of the studies have included sufficient taxon sampling to conclusively examine broad species patterns across the entire subfamily. Studies that have attempted to reconstruct higher‐level relationships of Cicindelinae have yielded conflicting results. Here, we present the first taxonomically comprehensive molecular phylogeny of Cicindelinae to date, with the goal of creating a framework for future studies focusing on this important insect lineage. We utilized all available published molecular data, generating a final concatenated dataset including 328 cicindeline species, with molecular data sampled from six protein‐coding gene fragments and three ribosomal gene fragments. Our maximum‐likelihood phylogenetic inferences recover Cicindelinae as sister to the wrinkled bark beetles of the subfamily Rhysodinae. This new phylogenetic hypothesis for Cicindelinae contradicts our current understanding of tiger beetle phylogenetic relationships, with several tribes, subtribes and genera being inferred as paraphyletic. Most notably, the tribe Manticorini is recovered nested within Platychilini including the genera Amblycheila Say, Omus Eschscholtz, Picnochile Motschulsky and Platychile Macleay. The tribe Megacephalini is recovered as paraphyletic due to the placement of the monophyletic subtribe Oxycheilina as sister to Cicindelini, whereas the monophyletic Megacephalina is inferred as sister to Oxycheilina, Cicindelini and Collyridini. The tribe Collyridini is paraphyletic with the subtribes Collyridina and Tricondylina in one clade, and Ctenostomina in a second one. The tribe Cicindelini is recovered as monophyletic although several genera are inferred as para‐ or polyphyletic. Our results provide a novel phylogenetic framework to revise the classification of tiger beetles and to encourage the generation of focused molecular datasets that will permit investigation of the evolutionary history of this lineage through space and time.     

Cytogenetic analysis of Australian tiger beetles (Coleoptera: Cicindelidae): chromosome number, sex-determining system and localization of rDNA genes

The chromosomes of four species of Australian tiger beetle are documented. The male meioformulae of three species of Cicindela (tribe Cicindelini) were determined as follow: Cicindela cardinalba Sumlin 1987, n = 10 + X₁ X₂ X₃Y; C. sp. ( saetigera group) and Cicindela gillesensis Hudson 1994, n = 11 + X₁ X₂ X₃Y. The male meioformula of Megacephala whelani Sumlin 1992 (tribe Megacephalini) is n = 12 + XY. Fluorescence in situ hybridization (FISH) was used to localize the 18S-28S ribosomal gene clusters on male mitotic and meiotic chromosomes and nuclei using a polymerase chain reaction-amplified ribosomal probe. Fluorescence in mitosis and meiotic first prophase stages in the three Cicindela species showed that rDNA genes are in two of the four chromosomes that form the sex vesicle. In M. whelani hybridization in mitosis and first metaphase stages indicate that rDNA genes are in three, medium- to small-sized, autosomal pairs. Silver staining of male meiotic nuclei reveals the presence of active nucleolar organizing regions (NORs) in the sex vesicle of the three species of Cicindela . The cytogenetic and evolutionary implications of these findings are discussed.     

Chromosomes of Trachypachus Motschulsky and the evolution of the ancestral adephagan karyotype (Coleoptera)

The family Trachypachidae is a critical group for understanding the evolution of the coleopteran suborder Adephaga. In this article, we report the first karyotypic data on Trachypachus showing a diploid number of 2n = 36 + X (meioformula n = 18 + X) and a single autosomal localization of the rDNA clusters. The evolutionary dynamics of this karyotype are discussed in the light of recent phylogenetic hypotheses of the order Coleoptera. We conclude that chromosome analysis supports a close relationship between trachypachids and the other Geadephaga and that a male karyotype with 36 + X chromosomes may well be considered ancestral for the whole suborder Adephaga.     

Karyotypic evolution and phylogeny of Mexican Passalidae (Coleoptera: Polyphaga: Scarabaeoidea)

The chromosomes of 26 taxa from Mexico of the tribes Passalini (three species) and Proculini (23 species) have been studied, increasing the karyotypically known species of the family Passalidae to 56. Karyotypic dynamism is high since the diploid number varies from 18 to 44 in the tribe Proculini. and from 25 to 31 in the tribe Passalini. In addition, supernumerary chromosomes, chromosome heteromorphism, translocations and possible sex multivalents have been found. These results contrast with the numerical conservatism found in related families of the superfamily Scarabaeoidea. However, both tribes are conservative with regard to sex determination, as all species of Proculini have male XY chromosomes whereas species of the Passalini have male XO chromosomes. It is postulated that differences in patterns shown by these two tribes are mainly due to population structure, because many species of Proculini are endemic to restricted areas of Meso and South America, favouring the settlement of karyotypic changes, whereas species of Passalini are distributed over large areas in the lowlands. It is also postulated that the ancestral karyotypic formula of the family is close to 12–14 pairs of autosomes although the ancestral male sex determination may be either XY or XO. At present only a weak relationship between morphological and karyotypic evolution has been found, which together with the marked numerical variability found within and between genera make it difficult to obtain phylogenetic conclusions from karyotypic results.     

Phylogenetic relationships of vespertilionid genera (Mammalia: Chiroptera) as revealed by karyological analysis

Banded karyotypes of 50 species belonging to 23 genera were analyzed. The diploid chromosome numbers ranged from 26 to 50. For karyotypic comparison we used a 44 chromosomes karyotype, consisting of 4 metacentric and 17 acrocentric autosomes, as “basic karyotype”. Almost all of its 25 autosomal arms could be identified in each individual karyotype. In 8 chromosomes, i. e. 1/2, 7, 11, 12, 13, 15, 23 and X, small inversions were detected. As a result, each of the chromosomes mentioned occurs in two states which differ slightly in their banding patterns. These were used as character states in the cladistic analysis together with other chromosomal rearrangements. The implications drawn from the cladogram obtained are: The Miniopterinae clearly belong to the Vespertilionidae but are the first to branch off from the common stem. The tribe Myotini should be raised to the rank of a subfamily. Within the largest subfamily Vespertilioninae, one autapomorphic chromosomal character was found for each of the tribes Vespertilionini and Pipistrellini. In addition, both tribes are distinguished from the other Vespertilioninae tribes by two synapomorphic features. These results allow for the first time an unequivocal classification of the systematically difficult “pipistrelloid” species. The species of the genus Pipistrellus (sensu Hill and Harrison 1987) are spread over the Pipistrelllni and Vespertilionini. We therefore suggest the splitting of this heterogenous genus into at least four genera. Only the members of the previous subgenus Pipistrellus constitute the genus Pipistrellus and belong to the Pipistrellini. The previous subgenera Hypsugo, Vespadelus and Falsistrellus, given generic rank in some recent studies, belong to the tribe Vespertilionini and are not closely related to Pipistrellus. For the genera Eptesicus and Hesperoptenus, which belong neither to the Vespertilionini nor to the Pipistrellini, the tribe Eptesicini was established. The phylogenetic relations of this tribe and the status of the presumably polyphyletic tribe “Nycticeiini” could not be solved.     

Genome size evolution in Sapindaceae at subfamily level: a case study of independence in relation to karyological and palynological traits

Sapindaceae s.l. is a moderately large family of trees, shrubs and lianas. Current knowledge on genome size and how it varies in this family is scarce. This research aims to characterize the DNA content in 39 species of Sapindaceae, mainly in tribe Paullinieae s.s., by the analysis of the variation in genome size relative to karyotypic and palynological features. Nuclear DNA amount was measured by flow cytometry, and linear regression analyses were conducted to analyse the relationship between genome size variation and various karyotypic and palynological features. Genome size varied nine‐fold among species, ranging from 1C = 0.305 pg (Lophostigma plumosum) to 2.710 pg (Cardiospermum heringeri). The low regression coefficients obtained suggest that genome size mainly varies independently of karyotypic and palynological features. With regard to karyotype evolution, the constant chromosome number but variable genome size in Houssayanthus, Paullinia and Serjania suggest that structural changes mainly caused by changes in the amounts of repetitive DNA are more important than numerical change. In contrast, in Cardiospermum and Urvillea, variation in chromosome number and genome size supports the suggestion that numerical and structural changes are important in the karyotype evolution of these genera. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 174 , 589–600.     

A cytogenetic study on three Chilean species of Chrysomelinae (Coleoptera, Chrysomelidae)

Three species of Chilean leaf beetles were chromosomally analyzed. The endemic Araucanomela wellingtonensis displays a male meioformula of 13 + Xyp with 2n = 28 chromosomes and an asymmetric karyotype with two large autosome pairs and 12 medium/small pairs of autosomes and sex-chromosomes, a diploid number which had not been found among the other species of the subtribe Paropsina sensu lato studied to date. Strichosa eburata presents a meioformula of 11 + Xyp, 2n = 24 chromosomes, as occurs in many species of chrysomelines belonging to different subtribes. Furthermore, Phaedon cyanopterum has a 16 + Xyp meioformula, that is 2n = 34 chromosomes, of small size mostly, also in agreement with the karyological findings obtained in all the other congeneric species so far examined. These cytogenetic data are discussed with respect to the previous ones in this subfamily and with other characters of taxonomic and evolutionary value.     

Mechanisms of chromosomal evolution and its possible relation to natural history characteristics in Ancistrus catfishes (Siluriformes: Loricariidae)

Ancistrus is the most speciose genus of the tribe Ancistrini, with 58 valid species and many yet to be described. Cytogenetic studies were conducted on five apparently undescribed species from the Amazon basin, which showed different diploid numbers: Ancistrus sp. Purus (2n = 34); Ancistrus sp. Macoari (2n = 46); Ancistrus sp. Dimona (2n = 52); Ancistrus sp. Vermelho (2n = 42) and Ancistrus sp. Trombetas (2n = 38). All species possessed only one pair of NOR‐carrying chromosomes, but with extensive variation in both the location on the chromosome as well as in the position of the ribosomal sites on the karyotype. The karyotypic evolution of Ancistrus species seems to be based on chromosomal rearrangements, with a tendency to a reduction of the diploid number. Two new instances of XX/XY sex chromosomes for Ancistrus species, based on the heteromorphism in the male karyotype, were also recorded. The large karyotypic diversity among Ancistrus species may be related to biological and behavioural characteristics of these fish that include microhabitat preferences, territoriality and specialized reproductive tactics. These characteristics may lead to a fast rate of fixation of chromosomal mutations and eventually speciation across the basin.     

Phylogenetic relationships within the tribe Janieae (Corallinales,Rhodophyta) based on molecular and morphological data: a reappraisal of Jania1

Generic boundaries among the genera Cheilosporum, Haliptilon, and Jania—currently referred to the tribe Janieae (Corallinaceae, Corallinales, Rhodophyta)—were reassessed. Phylogenetic relationships among 42 corallinoidean taxa were determined based on 26 anatomical characters and nuclear SSU rDNA sequence data for 11 species (with two duplicate plants) referred to the tribe Corallineae and 15 species referred to the tribe Janieae (two species of Cheilosporum, seven of Haliptilon, and six of Jania, with five duplicate plants). Results from our approach were consistent with the hypothesis that the tribe Janieae is monophyletic. Our data indicate, however, that Jania and Haliptilon as currently delimited are not monophyletic, and that Cheilosporum should not be recognized as an independent genus within the Janieae. Our data resolved two well‐supported biogeographic clades for the included Janieae, an Indian‐Pacific clade and a temperate North Atlantic clade. Among anatomical characters, reproductive structures reflected the evolution of the Janieae. Based on our results, three genera, Cheilosporum, Haliptilon, and Jania, should be merged into a single genus, with Jania having nomenclatural priority. We therefore propose new combinations where necessary of some species previously included in Cheilosporum and Haliptilon.     

Phylogeny,classification and evolution of the water scavenger beetle tribe Hydrobiusini inferred from morphology and molecules (Coleoptera: Hydrophilidae: Hydrophilinae)

The water scavenger beetle tribe Hydrobiusini contains 47 species in eight genera distributed worldwide. Most species of the tribe are aquatic, although several species are known to occur in waterfalls or tree mosses. Some members of the tribe are known to communicate via underwater stridulation. While recent morphological and molecular‐based phylogenies have affirmed the monophyly of the tribe as currently circumscribed, doubts remain about the monophyly of included genera. Here we use morphological and molecular data to infer a species‐level phylogeny of the Hydrobiusini. The monophyly of the tribe is decisively supported, as is the monophyly of most genera. The genus Hydrobius was found to be polyphyletic, and as a result the genus Limnohydrobius stat. rev. is removed from synonymy with Hydrobius, yielding three new combinations: L. melaenus comb.n. , L. orientalis comb.n. , and L. tumbius comb.n. Recent changes to the species‐level taxonomy of Hydrobius are reviewed. The morphology of the stridulatory apparatus has undergone a single remarkable transformation within the lineage, from a simple, unmodified pars stridens to one that is highly organized and complex. We present an updated key to genera, revised generic diagnoses and a list of the known distributions for all species within the tribe.     

POLLEN MORPHOLOGY OF THE TRIBE SWARTZIEAE (SUBFAMILY PAPILIONOIDEAE: LEGUMINOSAE). 1. INTRODUCTION AND ALL GENERA EXCLUDING ALDINA AND SWARTZIA

The pollen morphology of 9 of the 11 genera of the tribe Swartzieae is described together with that of Holocalyx and Cyathostegia, two genera recently removed from the tribe based on macromorphology. The pollen is small, spheroidal to subprolate, primarily tricolporate with a perforate tectum and generally typical of the Leguminosae. Nonetheless, many of the genera have distinctive pollen morphology. Baphiopsis is 6-colporate. The genera Harleyodendron, Lecointea and one species of Exostyles have supratectal spinules. Bocoa viridiflora has striate/rugulate ornamentation very distinct from the other species of the genus Bocoa. The exine stratification is varied but Candolleodendron has a very thick endexine and narrow foot layer. The pollen of African taxa does not differ significantly from that of South American taxa. Pollen morphology does not clarify the taxonomic relationships of the tribe and provides little evidence to assist in positioning Holocalyx and Cyathostegia.     

Genetic divergence in the tribe Electronini (Myctophidae)

To reveal evolutionary relationships and the rate of divergence of lanternfishes in the tribe Electronini (Myctophidae), analysis of sequence of cox 1 mtDNA fragment in three genera (12 species) of the tribe and in a closely related genus Myctophum (9 species) was performed. The results support the tribe monophyly; however, the divergence within it appeared to be not profound: genetic distances between genera and subgenera are smaller than between species within Myctophum and other genera of myctophids. In subgenera of Protomyctophum, species formation has obviously not completed since interspecies distances in them (0.5–4.7%) are comparable to estimates of intraspecific variation in other lanternfishes. Minimal genetic differences were found between species P. (Hierops) arcticum and P. (H.) subparallelum, whose ranges at the present time are isolated in the northern and southern hemispheres. After calibration of the molecular clock, the time of isolation of these two species can be dated to the Middle Pleistocene. Weak tribe diversification indicates its evolutionary youth, which can be related to the absence of efficient hydrographic barriers for isolation. The data obtained do not permit to consider tribe members as the most primitive in the family Myctophidae.     

Island species radiation and karyotypic stasis in <Emphasis Type="Italic">Pachycladon</Emphasis>allopolyploids

Background  

Pachycladon (Brassicaceae, tribe Camelineae) is a monophyletic genus of ten morphologically and ecogeographically differentiated, and presumably allopolyploid species occurring in the South Island of New Zealand and in Tasmania. All Pachycladon species possess ten chromosome pairs (2n = 20). The feasibility of comparative chromosome painting (CCP) in crucifer species allows the origin and genome evolution in this genus to be elucidated. We focus on the origin and genome evolution of Pachycladon as well as on its genomic relationship to other crucifer species, particularly to the allopolyploid Australian Camelineae taxa. As species radiation on islands is usually characterized by chromosomal stasis, i.e. uniformity of chromosome numbers/ploidy levels, the role of major karyotypic reshuffling during the island adaptive and species radiation in Pachycladon is investigated through whole-genome CCP analysis.     

Characterization of a family of tandemly repeated DNA sequences in Triticeae

The recombinant plasmid dpTa1 has an insert of relic wheat DNA that represents a family of tandemly organized DNA sequences with a monomeric length of approximately 340 bp. This insert was used to investigate the structural organization of this element in the genomes of 58 species within the tribe Triticeae and in 7 species representing other tribes of the Poaceae. The main characteristic of the genomic organization of dpTa1 is a classical ladder-type pattern which is typical for tandemly organized sequences. The dpTa1 sequence is present in all of the genomes of the Triticeae species examined and in 1 species from a closely related tribe (Bromus inermis, Bromeae). DNA from Hordelymus europaeus (Triticeae) did not hybridize under the standard conditions used in this study. Prolonged exposure was necessary to obtain a weak signal. Our data suggest that the dpTa1 family is quite old in evolutionary terms, probably more ancient than the tribe Triticeae. The dpTa1 sequence is more abundant in the D-genome of wheat than in other genomes in Triticeae. DNA from several species also have bands in addition to the tandem repeats. The dpTa1 sequence contains short direct and inverted subrepeats and is homologous to a tandemly repeated DNA sequence from Hordeum chilense.     

Molecular systematics of the butterfly tribe Preponini (Nymphalidae: Charaxinae)

The nymphalid butterfly tribe Preponini includes some of the Neotropical region's most spectacular and familiar butterflies, but the taxonomy of the group nevertheless remains unstable. Several recent studies of Nymphalidae phylogeny have suggested that both the tribe itself and several genera might not be monophyletic, but to date taxon sampling has not been sufficiently comprehensive to allow informed revision of the group's systematics. We therefore conducted the first complete species‐level phylogenetic study of the tribe to establish a firm higher classification. We used DNA sequence data from three genes, the two mitochondrial genes cytochrome oxidase subunits I and II (COI and COII), and the nuclear gene elongation factor‐1α (EF‐1α), to reconstruct the phylogeny of the tribe using maximum likelihood (ML), maximum parsimony (MP) and Bayesian inference (BI). We included 48 individuals representing the 22 recognised Preponini species, and an additional 25 out‐group taxa to explore taxonomic limits at different levels. Firstly, we found that Anaeomorpha splendida Rothschild never grouped with remaining Preponini, so that maintaining monophyly of the tribe requires the taxon to be excluded, and we thus reinstate the tribe Anaeomorphini stat.rev. Secondly, we investigated generic limits, in particular the relationship of Noreppa Rydon to Archaeoprepona Fruhstorfer, and that of Agrias Doubleday to Prepona Boisduval. The molecular results coupled with previous morphological studies suggest that Noreppa syn.n should be synonymised with Archaeoprepona, and that Agrias syn.n should be synonymised with Prepona. We found Prepona pheridamas (Cramer) to be sister to all other Prepona, and markedly divergent from them in both morphology and DNA sequences, suggesting the possibility that it should be placed in a separate genus. We also found a number of cases of significant DNA sequence divergence and paraphyly or polyphyly within putative species that require further taxonomic attention, including Prepona claudina (Godart) stat.n. and Prepona narcissus (Staudinger) stat.n., Prepona pylene Hewitson and Prepona deiphile (Godart). Future research should focus on a broader population sampling of widespread, polymorphic Preponini species to thoroughly revise the current species‐level taxonomy, thus creating a solid foundation for studies in ecology and conservation.     

Karyotypic characterization of three weevil species (Coleoptera: Curculionidae, Brachyderini)

Karyotypes of three species, Brachyderes incanus, Brachysomus setiger and Paophilus afflatus, belonging to the tribe Brachyderini, were studied using C-banding technique. The species share the same chromosome number 2n = 22 and meioformula n = 10+Xy(p) at all metaphase 1 plates of spermatid division. Some differences between karyotypes were observed in terms of centromere positions and C-band sizes. Most chromosomes are meta- or submetacentric and form a graded series in respect to length. The chromosomes resemble one another in having a rather small amount of heterochromatin restricted to the pericentromeric region and visible as dark stained blocks mainly during early stages of nuclear division. Only in Brachyderes incanus do larger bands occur at mitotic metaphase and diakinesis. These cytogenetic data are in agreement with karyological findings obtained in other species of Brachyderini so far examined.     

Phylogenetic Relationships of African Killifishes in the GeneraAphyosemionandFundulopanchaxInferred from Mitochondrial DNA Sequences

We have analyzed the phylogenetic relationships of 52 species representing all defined species groups (J. J. Scheel, 1990, Atlas of Killifishes of the Old World, 448 pp.) of the African aplocheiloid fish generaAphyosemionandFundulopanchaxin order to examine their interrelationships and to reveal trends of karyotypic evolution. The data set comprised 785 total nucleotides from the mitochondrial 12S rRNA and cytochromebgenes. The molecular-based topologies analyzed by both maximum parsimony and neighbor-joining support the monophyly of most previously defined species groups within these two killifish genera. The genusAphyosemionis monophyletic except for the nested position ofFundulopanchax kunzi(batesigroup; subgenusRaddaella) within this clade, suggesting that this taxon was improperly assigned toFundulopanchax.The remainingFundulopanchaxspecies sampled were supported as being monophyletic in most analyses. Relationships among the species groups in both genera were not as strongly supported, suggesting that further data will be required to resolve these relationships. Additional sampling from the 16S rRNA gene allowed further resolution of relationships withinFundulopanchax,more specifically identifying the nonannualscheeligroup as the basal lineage of this otherwise annual genus. Chromosomal evolution withinAphyosemionhas been episodic, with the evolution of a reducedn= 9–10 metacentric complement having occurred in multiple, independent lineages. Polarity of chromosomal reductions within theelegansspecies group appears to support previous hypotheses concerning mechanisms of karyotypic change within the genusAphyosemion.     

CHROMOSOME NUMBERS IN THE COMPOSITAE. XIV. LACTUCEAE

Reports of 150 original chromosome counts are recorded, including reports of 22 genera and 57 species and subspecific taxa in tribe Lactuceae. Also included are first reports for 12 specific or subspecific taxa. x = 9 appears to be the ancestral base of the tribe. Chromosome numbers are known for over 85% of the genera of the tribe and the frequency of polyploidy is ca. 23%, which is about one-half that of the angiosperms.