Higher phylogeny of zygaenid moths (Insecta: Lepidoptera) inferred from nuclear and mitochondrial sequence data and the evolution of larval cuticular cavities for chemical defence

Zygaenid moths are capable of releasing hydrogen cyanide in their defense by enzymatic break-down of cyanoglucosides, but only larvae of chalcosiine and zygaenine moths store cyanogenic compounds in cuticular cavities and thus are able to discharge defense droplets, which effectively deter potential predators. A previously proposed phylogeny of Zygaenidae hypothesized a sister group relationship of chalcosiine and zygaenine moths because of their similar larval defense system. Not all chalcosiine taxa possess cuticular cavities, however, and a comparable defense mechanism has been reported in larvae of the zygaenoid family Heterogynidae. Considering sequence data of seven molecular loci, the present study estimates the posterior probability of phylogenetic hypotheses explaining the occurrence of larval cuticular cavities. The molecular data confirm the previous exclusion of Himantopteridae from Zygaenidae and suggest their close affinity to Somabrachyidae. The sequence data also corroborate the recently proposed exclusion of the Phaudinae from the Zygaenidae, because this subfamily is recovered in a reasonably well supported species cluster consisting of members of the families Lacturidae, Limacodidae, Himantopteridae, and Somabrachyidae. We consequently agree to raise Phaudinae to family rank. Within Zygaenidae, the subfamilies Callizygaeninae, Chalcosiinae, and Procridinae most likely constitute a monophyletic group, which is sister to the Zygaeninae. Our results imply that cuticular cavities were probably present in the larvae of the most recent common ancestor of Zygaenidae. Heterogynidae cannot be confirmed as sister taxon to this family, but appear at the very first split of the Zygaenoidea, although with poor support. The specific pattern of taxa in the molecular phylogeny showing larval cuticular cavities opens the possibility that these structures could have been already present in the most recent common ancestor of the Zygaenoidea.     

The phylogenetic relationships of Chalcosiinae (Lepidoptera, Zygaenoidea, Zygaenidae)

The chalcosiine zygaenid moths constitute one of the most striking groups within the lower-ditrysian Lepidoptera, with highly diverse mimetic patterns, chemical defence systems, scent organs, copulatory mechanisms, hostplant utilization and diapause biology, plus a very disjunctive biogeographical pattern. In this paper we focus on the genus-level phylogenetics of this subfamily. A cladistic study was performed using 414 morphological and biochemical characters obtained from 411 species belonging to 186 species-groups of 73 genera plus 21 outgroups. Phylogenetic analysis using maximum parsimony leads to the following conclusions: (1) neither the current concept of Zygaenidae nor that of Chalcosiinae is monophyletic; (2) the previously proposed sister-group relationship of Zygaeninae + Chalcosiinae is rejected in favour of the relationship (Zygaeninae + ((Callizygaeninae + Cleoda ) + ( Heteropan + Chalcosiinae))); (3) except for the monobasic Aglaopini, none of the tribes sensu Alberti (1954 ) is monophyletic; (4) chalcosiine synapomorphies include structures of the chemical defence system, scent organs of adults and of the apodemal system of the male genitalia. A paired metathoracic androconial organ and a series of abdominal tergal corematal organs have been discovered, both being new to Lepidoptera. Due to highly homoplastic patterns in copulatory structures and wings that demonstrate significant sexual dimorphism, polymorphism and mimicry, 17 of the 69 'true' chalcosiine genera ( c . 25%) are shown to be either paraphyletic or polyphyletic. The present classification is therefore very misleading. Reductions of various parts of the male genitalia in some groups are accompanied by morphological and functional replacement involving the 8th abdominal segment. A prominent but convergent lock and key mechanism is revealed.  © 2005 The Linnean Society of London, Zoological Journal of the Linnean Society , 2005, 143 , 161–341.     

The ultrastructure of the cuticle of Nematoda

 The ultrastructure of the body cuticle in species of six of seven representative genera of Stilbonematinae (Eubostrichus, Catanema, Laxus, Robbea, Leptonemella, and Stilbonema) was investigated using SEM and TEM techniques. Additionally, one species of Spirinia (Spiriniinae) and one of Desmodora (Desmodorinae) were studied for outgroup comparison. The body cuticle of all investigated stilbonematids shows a consistent pattern composed of specific elements in a characteristic arrangement to each other. This pattern does not occur in Stilbonematinae alone, but also in Desmodorinae and Spiriniinae. Furthermore, a comparison within the Desmodorida reveals that this cuticular pattern apparently is present in the cuticle of representatives of Monoposthiidae, Epsilonematidae, and Draconematidae. The present results lead to the following conclusions: (1) the cuticle of Stilbonematinae contains no autapomorphic characters for this taxon, (2) there is a common cuticular pattern within the Desmodorida, and (3) this pattern is an autapomorphic character for the order Desmodorida. Accepted: 4 February 1996     

Extrafloral nectaries in the genus Macaranga (Euphorbiaceae) in Malaysia: comparative studies of their possible significance as predispositions for myrmecophytism

Some species of the paleotropical tree genus Macaranga (Euphorbiaceae) live in close association with ants. The genus comprises the full range of species from those not regularly inhabited by ants to obligate myrmecophytes. In Malaysia (Peninsular and Borneo) 23 of the 52 species are known to be ant-associated (44%). The simplest structural adaptation of plants to attract ants are extrafloral nectaries. We studied the distribution of extrafloral nectaries in the genus Macaranga to assess the significance of this character as a possible predisposition for the evolution of obligate myrmecophytism. All species have marginal glands on the leaves. However, only the glands of non- myrmecophytic species function as nectaries, whereas liquids secreted by these glands in myrmecophytic species did not contain sugar. Some non-myrmecophytic Macaranga and transitional Macaranga species in addition have extrafloral nectaries on the leaf blade near the petiole insertion. All obligatorily myrmecophytic Macaranga species, however, lack additional glands on the lamina. The non-myrmecophytic species are visited by a variety of different ant species, whereas myrmecophytic Macaranga are associated only with one specific ant-partner. Since these ants keep scale insects in the hollow stems, reduction of nectary production in ant-inhabited Macaranga seems to be biologically significant. We interpret this as a means of (a) saving the assimilates and (b) stabilization of maintenance of the association's specificity. Competition with other ant species for food rewards is avoided and thereby danger of weakening the protective function of the obligate ant- partner for the plant is reduced. A comparison with other euphorb species living in the same habitats as Macaranga showed that in genera in which extrafloral nectaries are widespread, no myrmecophytes have evolved. Possession of extrafloral nectaries does not appear to be essential for the development of symbiotic ant-plant interactions. Other predispositions such as nesting space might have played a more important role.     

Harmonic Hopping,and Both Punctuated and Gradual Evolution of Acoustic Characters in Selasphorus Hummingbird Tail-Feathers

Models of character evolution often assume a single mode of evolutionary change, such as continuous, or discrete. Here I provide an example in which a character exhibits both types of change. Hummingbirds in the genus Selasphorus produce sound with fluttering tail-feathers during courtship. The ancestral character state within Selasphorus is production of sound with an inner tail-feather, R2, in which the sound usually evolves gradually. Calliope and Allen''s Hummingbirds have evolved autapomorphic acoustic mechanisms that involve feather-feather interactions. I develop a source-filter model of these interactions. The ‘source’ comprises feather(s) that are both necessary and sufficient for sound production, and are aerodynamically coupled to neighboring feathers, which act as filters. Filters are unnecessary or insufficient for sound production, but may evolve to become sources. Allen''s Hummingbird has evolved to produce sound with two sources, one with feather R3, another frequency-modulated sound with R4, and their interaction frequencies. Allen''s R2 retains the ancestral character state, a ∼1 kHz “ghost” fundamental frequency masked by R3, which is revealed when R3 is experimentally removed. In the ancestor to Allen''s Hummingbird, the dominant frequency has ‘hopped’ to the second harmonic without passing through intermediate frequencies. This demonstrates that although the fundamental frequency of a communication sound may usually evolve gradually, occasional jumps from one character state to another can occur in a discrete fashion. Accordingly, mapping acoustic characters on a phylogeny may produce misleading results if the physical mechanism of production is not known.     

Current understanding of the development of sex attractant-based biocontrol in burnet moths

Burnet moths, also known as the family Zygaenidae, are a typical diurnal family of Lepidoptera. Some species are important insect pests in agriculture and forestry. The use of sex attractants is one of the most important measures in the integrated pest management of burnet moths because these attractants are highly efficient, do not cause pollution, and are convenient and harmless to natural enemies. The earliest information about sex pheromones in Zygaenidae was reported in 1972. To date, many studies have described sex pheromones and attractants of species in Zygaenidae. Here, we review a total of 22 sex attractant compositions identified for 57 species of Zygaenidae and compare the differences in the structures of sex attractants among these species. The application of sex attractants in seasonal monitoring showed that the main activity period of zygaenid moths is from May to September each year. However, there are few reports on the effects of sex attractants on mass trapping of members of the family Zygaenidae. The ratio and degradation of sex attractants and the trap color, shape and hanging height were considered the main factors influencing capture effectiveness. Directions for further study of burnet moths are also discussed.     

Notobaetis: A New Genus of Baetidae (Ephemeroptera) from South America

A new genus and species, Notobaetis penai, n. gen., n. sp., of Baetidae (Ephemeroptera), is described based upon reared specimens collected from the Cordoba Province of Argentina. This new genus appears to be phenetically intermediate between Baetis Leach and Centroptilum Eaton while possessing a unique combination of characters and one autapomorphic character.     

On the phylogenetic significance of two miocene zygaenid moths (insecta,lepidoptera)

Two Miocene zygaenid moths (Insecta, Lepidoptera), »Zygaena« miocaenica Reiss andZygaenites controversus BuRGEFF, both from the Schwäbiscne Alb in south-western Germany, are redescribed and illustrated in detail. Comparison with a cladogram of the Zygaeninae illustrates that they belong to two different genera, i. e. to two different evolutionary lines of the subfamily. Characters in the venation ofZygaenites controversus demonstrate its close relationship to the extant genusEpizygaenella. However, there are no specific characters in »Zygaena« miocaenica which would allow a precise assignment of that species, altough it clearly shows derived characters of the (Praezygaena + Reissita + Epizygaenella + Zygaena) group. This ist the most advanced subdivision of the Zygaeninae, which orginated in the Afrotropics, although the main radiation occurred in Eurasia.     

Morphology of the fifth sternal glands of Neotropical social wasps (Hymenoptera,Vespidae, Polistinae)

Glands play a central role in the social behavior of insects, and comparative analysis of their structure may provide insights about their evolution and their role in insect social biology. Here, we describe the glands of the fifth metasomal sternite of ten polistine wasps with different social behaviors. The glands of foragers of these species were studied using semithin sections of Araldite‐embedded tissue, and scanning electron microscopy. The structure of the glands observed was mostly similar to what has previously been reported for independent‐founder and swarm‐founder species, respectively. Scale‐shaped modifications in the fifth sternite are reported for the first time for members of the genera Synoeca and Epipona. Two previously unreported glands in the fifth sternite are also described. A more restrictive definition of the name Richards' gland is proposed, applying to the class 3 cells associated with scale modifications of the fifth sternite, based on structural differences between the glands observed, homology requirements, and functional evidence. Previous phylogenetic reconstructions of this character suggest that it appeared early in the Vespidae and disappeared four times, with a single reappearance in the genus Vespula. The restricted definition of Richards' gland suggests that this structure is an exclusive trait of the Epiponini, with two reversals.     

Das Triplett aus zwei Monophyla und einem Paraphylum als Baustein des phylogenetischen Systems

The triplet consisting of two monophyletic taxa and one paraphyletic taxon as constructive element of the phylogenetic system Evolution has produced very many novelties (apomorphies). Most of them are small and relatively inconstant, these are more or less indicative of the phylogenetic relationships between closely related species. They cannot be the constitutive character of a supraspecific taxon that exists since a long time and comprises many diversified species. Such a taxon of higher rank can only be characterized by an improbable, rare novelty that has developed only once and has been preserved in all descendent species. Two consecutive apomorphies of this persistent type (‘fixed apomorphies’) characterize three supraspecific taxa, the triplet “A”, “B” and “A minus B” (Fig. 1). The group “A minus B” is rejected in Hennig's theory because it is ‘paraphyletic’, but it is not an artefact created by the systematicist. It is an inevitable mathematical consequence of the differentiatison of the group “B” within the group “A”. Being the result of a subtraction, it is necessarily associated with the two monophyletic partners in the triplet, as it is delimited on one side by the synapomorphy of the group “A”, of which it is a part, and on the other side by the autapomorphy of the separate group “B”. Traditional classifications often include paraphyletic groupings that are inconsistent with phylogenetics, e. g. the Reptilia and the Apterygota. The fault in such cases is that these groups are extended beyond the limits of a triplet and cover more than a single interval between consecutive monophyletic taxa. Paraphyletic groups are admitted in the phylogenetic system only for bridging the gaps in our cladistic information. According to HENNIG'S theory, all supraspecific taxa should be arranged two by two as sister-groups originating from one ancestral species and comprising all descendents of that species. The fixed evolutionary novelties which characterize higher supraspecific taxa are, however, rare and scattered. It is highly improbable that they have developed in sister species, therefore the taxa marked by them cannot be sister-groups (except in very rare cases). In HENNIG'S earlier papers, e. g. in his system of Lepidoptera (1953: 46–49), the alleged ‘sister-groups' are, in reality, the groups “B” and “A minus B” of a triplet (see Fig. 2). In his revised concept (1957 and later), two autapomorphic groups which are most closely related in the recent fauna (“B” and “C” in Fig. 3) are called ‘sister-groups’. But these have originated independently from different ancestors in a plesiomorphic complex of extinct species and are more closely related to parts of this complex than to each other. True sister-groups (“Bx” and “Cx” in Fig. 4) would be formed if these related plesiomorphic species were included, but this extension of the ’backward‘ border of the taxon is not justified by synapomorphy (in the terms of logic, it is a ’metabasis‘), and it would make the classification of fossil species impossible, unless these show at least one synapomorphy with either “B” or “C”. In the system of the recent fauna the sister-groups are identical with the autapomorphic groups, because the plesiomorphic species are extinct. The natural system based on synapomorphies and autapomorphies is the triplet-system as outlined in Figure 6. It is not a new type of classification, but its theoretical foundation was missing, and precise instructions were needed for its use in phylogenetics. The information obtained by HENNIG'S method is entirely preserved in this system and can be retrieved from it, and both recent and extinct species can be classified together. The disadvantage of the triplet-system is that it contains twice as many taxa as HENNIG'S classification. This complexity will limit its use in the practice of taxonomy, but it may be simplified by transforming the system into a sequence of paraphyletic taxa terminating in a single monophylum.     

Arenophilic mantle glands in the Laternulidae (Bivalvia: Anomalodesmata) and their evolutionary significance

The mantle margins of several anomalodesmatans bear multicellular arenophilic glands, the mucoid secretions of which attach sand grains and other foreign particles to the outer surface of the periostracum. These glands have been recorded for many of the anomalodesmatan families and are used as a key morphological character in recent attempts to unravel the evolutionary relationships within the Anomalodesmata. The glands occur in Laternula elliptica, L. truncata, L. boschasina and L. marilina, discharging from the top of muscular papillae at the distal tip of the siphons. The secretions are laid down as threads organized in longitudinal lines along the length of the periostracum that covers the siphonal walls. This is the first record of arenophilic mantle glands in members of the Laternulidae, a finding that not only broadens our current knowledge of the family's morphology, but assists in the reconstruction of anomalodesmatan evolutionary history.     

Molecular anatomy of an endodermal gland: Investigations on mucus glycoproteins and cell turnover in Brunner's glands of didelphis verginiana using lectins and PCNA immunoreactivity

Brunner's glandsl are located in the submucosa of the proximal duodenum and are unique to mammalian species. The North American opossum (Didelphis virginiana) is generally regarded as a prototype marsupial that closely resembles fossil didelphids which can be placed at the beginning of mammalian evolution. The current investigation provided an opportunity for the analysis of secretory products from these glands in a species thought to be more closely related to earlier evolutionary forms. Extracts of Brunner's glands were subjected to SDS-PAGE and Western Blotting. The results indicate the presence of two high molecular weight PAS-positive glycoprotein bands. In addition to these two PAS-positive bands, several other glycoprotein bands were detected in the high molecular weight range that bind several lectins which typically recognize O-linked carbohydrates indicative of mucus type glycoproteins. The same lectins bind to glandular structures in tissue sections. Comparison of lectin binding sites with the pyloric glands of the stomach and duodenal goblet cells indicates that brunner's glands carbohydrate residues resemble those of the pyloric glands more closely than those of the duodenal goblet cells. The low cell turnover rate in brunner's glands is in contrast to the rapid turnover rate of goble cell precursors in the duodenal crypts. The mucus composition and the cell turnover rate correlate well with embryological data and suggest that Brunner's glands of Didelphis evolved from an epithelium more closely associated closely associated with the stomach than that of the duodenum as the topography of the gland would suggest. © Wiley-Liss, Inc.     

Correlation Between the Mesodermal Male Genital Ducts and the Spermatophore Structure in a Ditrysian Moth,Zygaena trifolii (Esper, 1783) (Insecta,Lepidoptera, Zygaenidae)

Abstract The mesodermal male genital system and the spermatophore of Zygaena trifolii (Insecta, Lepidoptera, Zygaenidae) have been studied histologically. Whereas the long unpaired duct, the ductus mesospermaticus, has been found to consist of at least six distinctly different regions, the tissues and secretions of the seminal vesicles and the accessory glands were found to be quite uniform. Several of the different regions are separated from each other by epithelial septa (constrictions). The external structure and the internal organization of the spermatophore are described. Based on histological comparisons, the correlation between the contents of the spermatophore and the different parts of the male system is given.     

Chromosomal evolution and distribution of telomeric repeats in golden moles (Chrysochloridae, Mammalia)

Golden moles (Chrysochloridae) are small, subterranean mammals endemic to sub-Saharan Africa that together with tenrecs constitute one of six orders in Afrotheria. Here we present a comprehensive karyotypic comparison among six species/subspecies of golden moles based on G-banding and chromosome painting. By expanding the species representation to include a further five species recently published in a companion paper, we were able to map the distribution of telomeric repeats in ten species/subspecies that are representative of six of the nine currently recognized genera. We conclude that: (i) the monophyly of Amblysomus is supported by the amplification of heterochromatin in several pericentric regions and one intrachromosomal rearrangement; (ii) A. hottentotus meesteri groups as sister to a clade that contains A. h. hottentotus, A. h. longiceps, A. h. pondoliae and A. robustus, an association that is underpinned by a shared intrachromosomal rearrangement and the detection of telomeric sequences in the centromeres of all chromosomes of the three A. hottentotus subspecies and A. robustus but, importantly, not in those of A. h. meesteri. These findings indicate an absence of gene flow suggesting that A. h. meesteri should be elevated to specific status. We hypothesize that the lack of gene flow may, in part, reflect hybrid dysgenesis resulting from abnormal meiotic segregation as a consequence of differences in the nature of the centromeric specific satellites; (iii) chromosomes 7 and 13 of Chrysochloris asiatica are fused in both Calcochloris obtusirostris and Eremitalpa granti, but that the position of the centromere in the fused chromosome differs in each species. This suggests that rather than being indicative of common ancestry, the fusion is more likely a convergent character which has arisen independently in each lineage. Furthermore our painting data show two centromeric shifts that are probably autapomorphic for C. obtusirostris. Finally, we conclude that (iv) golden moles are characterized by strong karyotypic conservatism but in marked contrast to the constrained rates of change exhibited by most species, A. robustus is unique in that three autapomorphic fissions define its evolutionary history, and hence the more extensive reshuffling of its genome.     

Unusual location and structure of female pheromone glands in Theresimima (= Ino) ampelophaga Bayle-Berelle (Lepidoptera : Zygaenidae)

Morphological and behavioural studies strongly suggest that the sex pheromone glands of female Theresimima (= Ino) ampelophaga (Lepidoptera : Zygaenidae) are situated on the anterior part of the 3rd–5th abdominal tergites. The glandular epithelium consists of 2 cell types: gland cells and wrapping cells. The gland cells have a central microvilli-lined cavity which is in contact with the lumen of the hair. These hairs are exposed during the calling behaviour, and the pheromone is probably given off via pores in the scale wall.     

Ultrastructural and phylogenetic assessment of wax glands in pit scales (Homoptera : Coccoidea)

TEM/SEM and computerized images of 5 wax glands for 3 type species of Coccoidea (Homoptera): Asterodiaspis variolosa (Asterolecaniidae), Cerococcus quercus (Cerococcidae) and Lecanodiaspis sardoa (Lecanodiaspididae) were studied. Their cuticular structures were compared with 142, 56, and 61 species in their respective families to determine relationships among pit scale taxa. Significant differences include: the morphology of the outer and inner ductule of the tubular duct gland, structure of the pores (8-shaped, multilocular and quinquelocular), and the absence or presence of cribriform plates and their structural variations. Three distinctive tubular duct shapes (asterform, ceroform and lecanoform) are common in pit scale species. Apomorphic characteristics of the asterform tubular ducts include an absence of the inner ductule and the progressive reduction of the outer ductule's diameter from the pore to its inner end. These characters easily separate asterolecaniids from the cerococcid-lecanodiaspidid lineage. The constricted lecanoform tubular ducts and the curved teeth on the rim at the inner end of the outer ductule in the ceroform tubular ducts are regarded as autapomorphic. The presence of 8-shaped pores is considered a plesiomorphic condition. Specific cuticular variations of the 8-shaped pores, characterizing familial taxa, include pores even with the surface in asterolecaniids, pores with raised walls in cerococcids, and bent pores in lecanodiaspidids. The dominant 8-shaped pore patterns in pit scales are those arranged in a marginal band in lecanodiaspidids, in a swirl-like pore pattern in the cerococcids, and in a marginal row in asterolecaniids. A divergent evolutionary trend is noted for the structure of the cribriform plate; they are with micro-orifices in cerococcids, but without micro-orifices in lecanodiaspidids. The former state is considered apomorphic. Cribriform plates arranged in clusters characterize the cerococcids, while plates in longitudinal rows characterize the lecanodiaspidids. These data confirm the concept that the pit scales constitute a paraphyletic group and the Asterolecaniidae, Cerococcidae and Lecanodiaspididae are monophyletic.     

Prey synchronize their vigilant behaviour with other group members

It is generally assumed that an individual of a prey species can benefit from an increase in the number of its group''s members by reducing its own investment in vigilance. But what behaviour should group members adopt in relation to both the risk of being preyed upon and the individual investment in vigilance? Most models assume that individuals scan independently of one another. It is generally argued that it is more profitable for each group member owing to the cost that coordination of individual scans in non-overlapping bouts of vigilance would require. We studied the relationships between both individual and collective vigilance and group size in Defassa waterbuck, Kobus ellipsiprymnus defassa, in a population living under a predation risk. Our results confirmed that the proportion of time an individual spent in vigilance decreased with group size. However, the time during which at least one individual in the group scanned the environment (collective vigilance) increased. Analyses showed that individuals neither coordinated their scanning in an asynchronous way nor scanned independently of one another. On the contrary, scanning and non-scanning bouts were synchronized between group members, producing waves of collective vigilance. We claim that these waves are triggered by allelomimetic effects i.e. they are a phenomenon produced by an individual copying its neighbour''s behaviour.     

Species boundaries within Saprolegnia (Saprolegniales, Oomycota) based on morphological and DNA sequence data

Saprolegnia is a common and widespread genus of Oomycetes, however species identifications are difficult and uncertain. To test whether keys based on morphological characters could identify species as determined by molecular characters we determined partial DNA sequences for the 28S rRNA gene and the complete internal transcribed spacer (ITS) region for 55 isolates belonging to Saprolegnia and one isolate of Protoachlya hypogyna that exhibited saprolegnoid zoospore discharge in water culture. Phylogenetic analyses of the combined sequence data yielded 10 robustly supported clades that probably represent separate species. Morphological analyses of all isolates revealed that each DNA-based clade could be delimited from others by autapomorphic or unique combinations of morphological character states but not without employing several features previously not used at the species level. Taxonomic implications of these results are discussed and recommendations for less equivocal characterization of new Saprolegnia species are made.     

Epidermal keratinocyte stem cells: their maintenance and regulation

The epidermis is a stratified epithelium consisting of inter follicular regions and appendages (hair follicles, sweat glands, sebaceous glands). The dominant cell type (the keratinocyte) is arranged in groups of cells termed epidermal proliferative units (EPUs), and one centrally-located clonogenic stem cell is ultimately responsible for replacing the remainder of the cells in the unit. Evidence is reviewed which indicates that the epidermal Langerhan's cell (ELC), and the cells comprising the dermis, may modify the keratinocyte microenvironment to create stem cell ‘niches’ and cellular diversity within the basal layer.