Association of insect life stages using DNA sequences: the larvae of Philodytes umbrinus (Motschulsky) (Coleoptera: Dytiscidae)

Abstract. Insect life stages are known imperfectly in many cases, and classifications are based often on only one or a few semaphoronts of a species. This is unfortunate as information in alternative life stages often is useful for scientific study. Although recent examples of DNA in taxonomy have emphasized the identification of indistinguishable species, such sequence data facilitate the association of life history stages and hold considerable promise in phylogenetic analysis, evolutionary studies, diagnostics, etc. These concepts are discussed here and an example is provided from diving beetles (Dytiscidae: Coleoptera). Three unknown larval specimens of an apparent species of Laccophilinae collected in Namibia were associated with the species Philodytes umbrinus (Motschulsky) using DNA sequence data. An 806-bp portion of the gene cytochrome oxidase I was sequenced from the unknown larvae. Several identified adult specimens of species of Laccophilinae from Namibia were also sequenced, including two P. umbrinus specimens and specimens from four Laccophilus Leach species. Additional species of Laccophilus from other areas of the world also were sequenced, as were specimens of Agabetes acuductus (Harris), Australphilus saltus Watts, Neptosternus boukali Hendrich & Balke and a species of Laccodytes Régimbart. Parsimony analysis resulted in two most parsimonious trees with the unknown larva unambiguously resolved in a group with both adult specimens of P. umbrinus (bootstrap value = 100%). The average pairwise p-distance between the unknown larva and adult P. umbrinus specimens averaged 0.09% (0–0.14%), compared with an average divergence between other conspecifics in the analysis of 0.24% (0–0.82%) and an overall average divergence between species of 13.49% (1.90–19.86%). Based on this, the unknown larvae were assigned to P. umbrinus. The larvae are diagnosed and described and their relationship with other Laccophilinae is discussed.     

<Emphasis Type="Italic">Paragonimus skrjabini</Emphasis> Chen, 1959 (Digenea: Paragonimidae) and related species in eastern Asia: a combined molecular and morphological approach to identification and taxonomy

A molecular and morphometric investigation is reported on the species complex of mammalian lung.ukes of which Paragonimus skrjabini Chen, 1959 and P.miyazakii Kamo, Nishida, Hatsushika & Tomimura, 1961 are the best-known examples. This species complex (here called the P.skrjabini complex) is shown to be monophyletic using DNA sequences from the nuclear ITS2 region and the mitochondrial cox1gene. The latter marker permits the discrimination of populations, some previously named as distinct species, from various geographical locations in China and Japan. Morphometric analysis of a number of variables accords remarkably closely with the molecular results. Main findings are that (1) nominal P.skrjabini from Fujian Province in eastern China is phylogenetically very close to P.miyazakii from Japan. It is proposed that both taxa should be referred to the same subspecies as P.skrjabini miyazakii. (2) Populations from Guangdong Province, Guangxi Zhuang Autonomous Region, Sichuan and Hubei should be referred to P.skrjabini skrjabini. Specimens of P.skrjabini from the Yangtze basin in Sichuan and Hubei are remarkably similar to one another according to genetic and morphometric data, but are not so different from Guangdong populations that they should be referred to a different subspecies at this stage. Specimens of P.skrjabini from Yunnan have not been assigned to a subspecies at this stage. (3) Partly following earlier studies, we regard the following as synonyms of P.skrjabini: P.miyazakii (reduced to subspecific status); P.szechuanensis Chung & Tsao, 1962 (probably belongs to P.s.skrjabini); and P.hueitungensis Chung, Hsu, Ho, Kao, Shao, Chiu, Pi, Liu, Ouyang, Shen, Yi & Yao, 1975 (probably belongs to P.s.skrjabini ); P.veocularis (Chen & Li, 1979) (specimens from Fujian regarded as belonging to this species should be referred to P.skrjabini miyazakii: the subspecific status of specimens from the type-locality in northern Sichuan is unclear). A number of questions remain unresolved. The name P. hokuoensis Ho & Chung, 1964 was proposed for two individual metacercariae of distinctive appearance from southern Yunnan. DNA sequences from very similar metacercariae from the same locality place this nominal species within, or sister to, the P. skrjabini complex. As yet, nothing is known regarding adult morphology or biology of this taxon and we retain it here as a distinct species. P. heterorchis (Zhou, Pang & Hsiang, 1982) might be a synonym of P. skrjabini: the form of the metacercaria provides evidence against this view and further work is required. P. macrorchis Chen, 1962 has probably been confused with P. skrjabiniin China. Within China, the former probably occurs only on Hainan Island, although P. fukienensis Tang & Tang, 1962 from Fujian Province could be a synonym.     

A revision of the scale insect genera Etiennea and Platysaissetia (Homoptera: Coccidae) with particular reference to Africa

Abstract. From a study of P.castilloae (Cockerell), the type species of Platysaissetia Cockerell, it is considered that none of the other species presently placed in Platysaissetia are congeneric. All known species from Africa (and one from British Guiana) are redescribed and are here transferred to the genus Etiennea Matile-Ferrero; one species is synonymized and twelve species are described as new. Lectotypes have been designated for Etiennea kellyi (Brain) and E.montrichardiae (Newstead). Although not considered to be congeneric with P.castilloae, P.tsaratananae Mamet and P.crustuliforme (Green) are left in Platysaissetia.     

Structure and function of adipokinetic hormones of the large white butterfly Pieris brassicae

The large white butterfly Pieris brassicae L. (also called cabbage white) is very common in Europe, Asia and the northern region of Africa, and has also been found in South Africa during approximately the last 20 years. The species is considered a pest insect, with larvae attacking brassicaceous crops. The adult is a strong migratory flyer and new territory can be infested this way. As a first step to investigate methods for combating this pest species, the present study aims to determine the complement of adipokinetic peptides, here generically referred to as adipokinetic hormones (AKHs), which are required to regulate the mobilization of fuels for insect flight. Biological assays, as well as mass spectrometry, reveal information about the presence, structure and function of AKHs in P. brassicae: a methanolic extract of the corpora cardiaca has hypertrehalosaemic activity in cockroaches, does not cause hyperlipaemia in locusts, and has adipokinetic activity in P. brassicae itself. Liquid‐chromatography electrospray ion trap mass spectrometry reveals three peptides that can be associated with the AKH family: the non‐amidated undecapeptide Vanca‐AKH (pELTFTSSWGGK‐OH), the nonapeptide Manse‐AKH (pELTFTSSWG amide) and the novel octapeptide Piebr‐AKH (pELTFSSGW amide). Sequence confirmation of all three assigned structures is obtained from matching mass spectrometry spectra from synthetic and native peptides. Moreover, the synthetic peptides Manse‐AKH and Piebr‐AKH have significant hyperlipaemic (=adipokinetic) activity when injected into newly‐emerged adult cabbage white butterflies. The non‐amidated Vanca‐AKH is, apparently, incompletely processed Manse‐AKH without hormonal activity. Simulated dispersal flight is able to release AKHs, as indicated by the higher concentration of lipids in the haemolymph of adult P. brassicae after activity and rest periods.     

Observations on the morphology and life-cycle of Procerovum varium (Onji & Nishio, 1916) (Trematoda: Heterophyidae)

The life-cycle of Procerovum varium (Digenea: Haplorchiinae) was studied experimentally and the morphology of stages in the life-cycle has been described and illustrated. Infections with adult flukes were found in the pond heron Ardeola grayii and heavy infections with metacercariae were found, attached to the liver of the fish Oryzias melastigma (Oryziatidae) occurring in a freshwater stream situated in Visakhapatnam, India. The cercariae developing in the snail Thiara tuberculatapossessed typical haplorchiine features and were characterised by the presence of numerous cystogenous glands. Early stages of metacercarial development occurred free in the muscles of the fish intermediate host. The larvae reached the liver at 5 days post-infection, encystment commenced 2 days later and 15-day-old metacercariae were found to be infective to chicks, ducks and mice that served as suitable experimental hosts. The adult flukes obtained from natural and experimental infections showed many intraspecific variations, especially in the size and shape of the expulsor which depend on the quantity of sperm it contains. The validity of various species described in the genus and differentiated on the basis of differences in the size of the expulsor has been examined. It is concluded that only three species of the genus, namely P. varium, P. cheni and P. calderoni, are valid. "P. sisoni" of Chen (1949) is confirmed as a synonym of P. varium. P. varium is reported for the first time from India.     

Species of the genus Psoroptes (Acari: Psoroptidae): A taxonomic consideration

The biosystematic status of mite species belonging to the genus Psoroptes Gervais, 1841 is difficult to determine by phenotypic methods and has been subject to taxonomic revisions and ongoing debate. At present, the existence of five species, P. cuniculi (Delafond, 1859), P. ovis (Hering, 1838), P. equi (Hering, 1838), P. cervinus Ward, 1915 and P. natalensis Hirst, 1919, is generally accepted. This classification is based mainly on the host species, the localization of the mites on their hosts and morphological characters of male mites. However, a critical review of the literature indicates that the features used to discriminate between the five species are not unequivocal: (a) the localization of mite populations on host animals is not completely strict, (b) the lengths of the outer opisthosomal setae of male mites, which are the main morphological features used for species discrimination, overlap between the five postulated species, and (c) host specificity cannot be deduced from results of transfer experiments. Rather, conspecificity of the members of the genus Psoroptes has to be presumed which is supported by molecular genetic analyses. On these grounds and on rules of priority P. cervinus Ward, 1915, P. cuniculi (Delafond, 1859), P. natalensis Hirst, 1919 and P. ovis (Hering, 1838) are seen as synonyms of P. equi (Hering, 1838).     

Petraliellidae Harmer, 1957 (Bryozoa: Cheilostomata) from Queensland,Australia

Several species of the family Petraliellidae were first described from the coast of Queensland, including the type species of Sinupetraliella, S. litoralis. These species are redescribed from type, or topotype specimens, and include Petraliella concinna, which has not been certainly found since its introduction in 1891; lectotype material is designated for P. buski and P. magna. Six other species are described from Queensland, P. crassocirca, P. dentilabris, P. dorsiporosa, Mucropetraliella bennetti, M. serrata and M. tuberosa. The species Mucropetraliella tuberosa is a new addition to the Queensland fauna and is described and illustrated here for the first time since its introduction in 1884. The characters of the family Petraliellidae are briefly discussed and the genera to which the Queensland species are assigned is reviewed. A taxonomic key to the ten Queensland petraliellid species described is also provided.     

PCR-based,rapid diagnosis of parasitism of Lygus spp. (Hemiptera: Miridae) by Peristenus relictus (Hymenoptera: Braconidae)

Several species of Lygus (Heteroptera: Miridae: Mirini) are serious crop pests in North America where their parasitism rate by native nymphal parasitoids is generally lower than in Europe. Peristenus relictus (Ruthe) (formerly P. stygicus Loan) (Hymenoptera: Braconidae: Euphorinae) is the predominant nymphal parasitoid of several Lygus spp. in the warm Mediterranean region and has been a candidate for introduction against Lygus hesperus Knight and L. lineolaris (Palisot de Beauvois) in the southern US. We report a rapid, sensitive, and specific PCR-based assay for diagnosis of P. relictus immature stages within Lygus nymphs that entails three steps: DNA extraction, PCR of the partial mitochondrial COI gene, and agarose gel electrophoresis. The PCR-based methodology is species-specific because the target DNA of other sympatric, congeneric species was not amplified with use of the primers developed for P. relictus diagnosis. The sensitivity of the PCR method, assessed through spike tests, was established by the detection of a ratio of 1:10,000 P. relictus DNA to Lygus DNA. Molecular diagnosis of parasitism of field collected nymphs is achievable in one day, eliminating the need to rear nymphs to obtain adult parasitoids for morphological identification.     

Eye ontogeny in Pleurodema bufoninum: A comparison with Pleurodema somuncurense (Anura,Leptodactylidae)

Vision is one of the main sensory systems in amphibians, and the eye structure is highly associated with habitat conditions. The ontogeny, as well as the adult structure, of the eye has been studied in only a few species. The life change after metamorphosis is accompanied by changes in the visual environment. The aim of this work is to describe the eye ontogeny of Pleurodema bufoninum and to compare it with that of Pleurodema somuncurense . Specimens of both Pleurodema species were processed for histology analysis at different stages of development, including the tadpole, postmetamorphic, and adult forms. Eyes in both Pleurodema species are composed of the 3 tunics, tunica fibrosa , tunica vasculosa , and tunica interna , and the lens. Additionally, in both, the iris presents a projection on its dorsal and ventral free ends that screens the cornea. This structure has been reported in the eye of several anuran species and is called the umbraculum, meniscus or pupillary nodule. Our results show that the structures related to light capture (retina and lens) appear early in larval life, while the components of the terrestrial‐life eye (scleral cartilage, specialized cornea, eyelids, nictitating membrane, and Harderian's gland) do not develop until the metamorphic climax, when the tadpole leaves the water. The adult eyes of P. bufoninum and P. somuncurense are very similar in structure and development.     

Phylogeny of Physarida (Amoebozoa,Myxogastria) Based on the Small‐Subunit Ribosomal RNA Gene,Redefinition of Physarum pusillum s. str. and Reinstatement of P. gravidum Morgan

Myxomycetes (also called Myxogastria or colloquially, slime molds) are worldwide occurring soil amoeboflagellates. Among Amoebozoa, they have the notable characteristic to form, during their life cycle, macroscopic fruiting bodies, that will ultimately release spores. Some 1,000 species have been described, based on the macroscopic and microscopic characteristics of their fruiting bodies. We were interested in Physarum pusillum (Berk. & M.A. Curtis) G. Lister, a very common species described with two variants, each bearing such morphological differences that they could represent two distinct species. In order to test this, we observed key characters in a large selection of specimens attributed to P.  pusillum, to its synonyms (in particular Physarum gravidum), and to related species. In addition, the small‐subunit ribosomal RNA gene was obtained from seven of these specimens. Based on these data, we provide a comprehensive phylogeny of the order Physarida (Eukaryota: Amoebozoa: Conosa: Macromycetozoa: Fuscisporidia). Morphology and phylogeny together support the reinstatement of P. gravidum Morgan 1896 with a neotype here designated, distinct from P. pusillum, here redefined.     

Comparing longevity of Pseudacteon species of different sizes: effect of sugar feeding

Sugar feeding is known to enhance lifespan in many parasitoid species. Several species of phorid flies in the genus Pseudacteon (Diptera: Phoridae) have been introduced in the Southern U.S.A. for the biological control of imported fire ants of Solenopsis species (Hymenoptera: Formicidae). Pseudacteon species are short‐lived flies and little is known about their nutritional ecology. Results from previous studies in our laboratory show that one of the introduced species, Pseudacteon tricuspis Borgmeier, is capable of feeding on sugar sources with a significant increase in lifespan. However, in Pseudacteon phorid flies, the degree to which sugar feeding can enhance the lifespan of other species is not clear, nor is it known whether there is a relationship between body size and longevity. In the present study, the effect of sugar feeding on the survival of three phorid fly species of different body sizes, Pseudacteon cultellatus Borgmeier, Pseudacteon curvatus Borgmeier and Pseudacteon obtusus Borgmeier, is investigated. For all three species, flies continuously provided with 40% or 20% sucrose solution live longer than individuals provided water only or 40% sucrose solution only on the first day of adult life. However, the degree to which sugar feeding enhanced longevity varies by species and is highest for P. obtusus. The data also indicate that longevity in Pseudacteon phorid flies is related to body size: the largest species (P. obtusus) lives significantly longer than the smaller species (P. cultellatus and P. curvatus). These results suggest that adults of all three Pseudacteon species are capable of feeding on sucrose solution and that sugar feeding can enhance their longevity. The significance of these results is discussed in relation to the field performance of Pseudacteon phorid flies as biological control agents of imported fire ants.     

Comparative growth in the postnatal skull of the extant North American turtle Pseudemys texana (Testudinoidea: Emydidae)

Bever, G.S. 2007. Comparative growth in the postnatal skull of the extant North American turtle Pseudemys texana (Testudinoidea: Emydidae). —Acta Zoologica (Stockholm) 88 : 000–000 Postnatal growth is one of the many aspects of developmental morphology that remains distinctly understudied in reptiles. Variation and ontogenetic scaling within the skull of the extant emydid turtle, Pseudemys texana is described based on 25 continuous characters. Results indicate that skull shape in this species changes little during postnatal growth relative to the only cryptodire taxa for which comparable datasets are available (Apalone ferox and Sternotherus odoratus). This relative lack of change results in the paedomorphic retention of a largely juvenile appearance in the adult form of P. texana. The skulls of males and females, despite the presence of distinct sexual dimorphism in size, grow with similar scaling patterns, and the few observed differences appear to reflect alteration of the male growth trajectory. Comparisons with A. ferox and S. odoratus reveal a number of similarities and differences that are here interpreted within a phylogenetic context. These preliminary hypotheses constitute predictive statements that phylogenetically bracket the majority of extant cryptodire species and provide baseline comparative data that are necessary for the future recognition of apomorphic transformations. Plasticity of ontogenetic scaling as a response to the homeostatic needs and behaviour of individuals commonly is evoked as a limitation of ontogenetic scaling as a means to inform phylogenetic studies. These evocations are largely unfounded considering that variability itself can evolve and thus be phylogenetically informative.     

Relative growth of the limbs and trunk in sifakas: Heterochronic,ecological, and functional considerations

Limb, trunk, and body weight measurements were obtained for growth series of Milne-Edwards's diademed sifaka, Propithecus diadema edwardsi, and the golden-crowned sifaka, Propithecus tattersalli. Similar measures were obtained also for primarily adults of two subspecies of the western sifaka: Propithecus verreauxi coquereli, Coquerel's sifaka, and Propithecus verreauxi verreauxi, Verreaux's sifaka. Ontogenetic series for the larger-bodied P. d. edwardsi and the smaller-bodied P. tattersalli were compared to evaluate whether species-level differences in body proportions result from the differential extension of common patterns of relative growth. In bivariate plots, both subspecies of P. verreauxi were included to examine whether these taxa also lie along a growth trajectory common to all sifakas. Analyses of the data indicate that postcranial proportions for sifakas are ontogenetically scaled, much as demonstrated previously with cranial dimensions for all three species (Ravosa, 1992). As such, P. d. edwardsi apparently develops larger overall size primarily by growing at a faster rate, but not for a longer duration of time, than P. tattersalli and P. verreauxi; this is similar to results based on cranial data. A consideration of Malagasy lemur ecology suggests that regional differences in forage quality and resource availability have strongly influenced the evolutionary development of body-size variation in sifakas. On one hand, the rainforest environment of P. d. edwardsi imposes greater selective pressures for larger body size than the dry-forest environment of P. tattersalli and P. v. coquereli, or the semi-arid climate of P. v. verreauxi. On the other hand, as progressively smaller-bodied adult sifakas are located in the east, west, and northwest, this apparently supports suggestions that adult body size is set by dry-season constraints on food quality and distribution (i. e., smaller taxa are located in more seasonal habitats such as the west and northeast). Moreover, the fact that body-size differentiation occurs primarily via differences in growth rate is also due apparently to differences in resource seasonality (and juvenile mortality risk in turn) between the eastern rainforest and the more temperate northeast and west. Most scaling coefficients for both arm and leg growth range from slight negative allometry to slight positive allometry. Given the low intermembral index for sifakas, which is also an adaptation for propulsive hindlimb-dominated jumping, this suggests that differences in adult limb proportions are largely set prenatally rather than being achieved via higher rates of postnatal hindlimb growth. Our analyses further indicate that the larger-bodied P. d. edwardsi has a higher adult intermembral index than P. tattersalli and P. verreauxi, thus supporting the allometric argument regarding the interspecific scaling of limb proportions in arboreal primates which employ vertical postures (Cartmill, 1974,1985; Jungers, 1978, 1985). Lastly, additional analyses indicate that P. d. edwardsi exhibits significant sexual dimorphism where adult females are larger than adult males in about one-third of all adult comparisons, whereas P. tattersalli exhibits significant sexual dimorphism in about one-fifth of all adult comparisons. Among western sifakas, adult P. v. coquereli exhibit significant sex dimorphism in about one-third of all comparisons, whereas adult P. v. uerreauxi show no significant differences between the sexes. Given that all taxa are ontogenetically scaled, this suggests that sexual dimorphism develops via such processes as well. Interestingly, our data indicate that sex dimorphism is allometric, with larger-bodied taxa like P. d. edwardsi being more dimorphic.     

A revision of the <Emphasis Type="Italic">Paraulopus oblongus</Emphasis> group (Aulopiformes: Paraulopidae) with description of a new species

 Paraulopus brevirostris, P. filamentosus, P. japonicus, P. legandi, P. maculatus, and P. oblongus are redescribed. All species are included in the Paraulopus oblongus group, characterized by having no supraocular ridge, 2.5–3.5 scales above the lateral line, and small adult body size (70–150 mm SL). In addition, P. atripes, from the Indian Ocean, is described as a new species of the P. oblongus group, being defined by the following combination of characters: small antrorse dentary process on chin; eye directed laterally; pelvic fin black in males; caudal fin white; 32–34 gill rakers; 45–46 pored lateral-line scales; and 3.5 scales below lateral line. Paraulopus albimaculatus is regarded as a junior synonym of P. brevirostris, based on examination of type specimens. A key to species in the P. oblongus group is included. Received: September 2, 2002 / Revised: January 31, 2003 / Accepted: February 17, 2003     

Systematics, biogeography and host plant associations of the Pseudomyrmex viduus group (Hymenoptera: Formicidae), Triplaris- and Tac/Mga/t-inhabiting ants

The Pseudomyrmex viduus group is a Neotropical clade of arboreal ants containing 13 species, of which three are newly described here: P. insuavis, P. ultrix, and P. vitabilis. Most species inhabit the domatia of specialized ant-plants. The ants keep brood and scale insects (Coccoidea) in the plant cavities, and defend their nest sites much more aggressively than do generalist species of Pseudomyrmex nesting in dead twigs. Five species are obligate associates of trees in the genus Triplaris (Polygonaceae) and five taxa are restricted to Tachigali (Fabaceae: Caesalpinioideae). One species, P. viduus (F. Smith), is much less host-specific, inhabiting Cordia, Coussapoa, Ocotea, Pseudobombax, Pterocarpus, Sapium, Triplaris and other myrmecophytes. Nothing is known about the biology of P. vitabilis, a close relative of/?viduus. The last member of the species group, P. kuenckeli (Emery), is a non-specialist but aggressive species that nests in dead branches of various plants. A cladistic analysis indicates that this aggressive behaviour evolved before obligate associations with specialized ant-plants, and that the Triplaris and Tachigali inhabitants each form their own clade. P. viduus, which is nested within the Triplaris-associated clade, suggests a possible model for host plant evolution in these ants wherein shifts from one ant-plant to another involve an intermediate phase of expanded host plant use. At least nine other Pseudomyrmex species, from two different species groups (not closely related to the P. viduus group), have evolved specialized associations with Triplaris or Tachigali including five new species: P. crudelis, P. deminutus, P. eculeus. P. ferox and P. hospitalis. Although the P. viduus group is centred in the Amazon basin, the geographic ranges of most species do not coincide with the Pleistocene forest refugia proposed by Haffer and others. A consideration of the phylogenetic relationships, distribution patterns, and host plant specificity of the ants indicates that much of the diversification of the P. viduus group occurred before the Pleistocene, and that the interactions with Triplaris and Tachigali plants are also of Tertiary origin.     

Protrellus dixoni n. sp. (Nematoda: Thelastomatidae) from the cockroach Drymaplaneta variegata

Abstract

In New Zealand, Protrellus dixoni n. sp. occurs in the hind gut of the introduced Australian cockroach Drymaplaneta variegata. It is distinguished from all other Protrellus species by its morphology. Females have a well developed muscular ovijector with a sphincter, and a conical tail with a filiform projection; males have four, not three, tail papillae. The taxonomic history of the genus is discussed and the generic diagnosis emended. New combinations include Protrellus eurycotesi, P. ischnopterae, P. ituana, and P. gurri. The population structure of P. dixoni in adult hosts is described: there are never more than one adult male, and usually few (one to three) adult females per host.     

CHROMOSOMAL ANALYSES AND THE ORIGIN OF ALLOPOLYPLOID POLYPODIUM VIRGINIANUM (POLYPODIACEAE)

Polypodium virginianum L. has been treated as a morphologically and chromosomally variable species. Recently, however, two species have been named from within the P. virginianum complex. Chromosome counts reported here confirmed that these segregate species, P. appalachianum Haufler and Windham and P. sibiricum Siplivinskij, are diploids with n = 37. It was further hypothesized that hybridization between these diploids initiated the allotetraploid P. virginianum. Naturally occurring, sterile triploid plants obtained from habitats in which the putative allotetraploid was sympatric with its presumed diploid progenitors have been verified isozymically as backcross hybrids. Meiotic chromosomal behavior showed that the triploids contained univalents and bivalents in nearly equal numbers, confirming that tetraploid P. virginianum contained one genome from each diploid. Observation of three to six trivalents in triploid preparations suggested that the diploids are still closely related genetically. This hypothesis was supported by isozymic studies which found a genetic identity of 0.610 between P. appalachianum and P. sibiricum, a value much higher than the average identity for congeneric fern species studied to date (0.33) and higher than any other species pair in the P. vulgare L. complex (average I = 0.307). These chromosomal and isozymic data help to resolve a biosystematic riddle that has interested pteridologists for over 40 years.     

Diapause in the seasonal cycle of stink bugs (Heteroptera,Pentatomidae) from the Temperate Zone

The paper reviews the data on diapause and related phenomena in stink bugs (Heteroptera: Pentatomidae). Using stink bugs as examples, the consecutive stages of the complex dynamic process of diapause (such as diapause preparation, induction, initiation, maintenance, termination, post-diapause quiescence, and resumption of direct development) are described and discussed. Out of 43 pentatomid species studied in relation to diapause in the Temperate Zone up to date, the majority (38 species) overwinter as adults, two species—as eggs, and another two species—as nymphs. Pentatoma rufipes is believed to be able to overwinter at different stages of its life cycle. Less than 5 % of pentatomid species are probably able to overwinter twice. Only five species have obligate diapause, others have the facultative one. Day-length and temperature are the main diapause inducing factors in the majority of species. The role of food in the control of seasonal development is essential in the pentatomid species feeding on plant seeds. In different species, different stages are sensitive to day-length. Some pentatomids retain sensitivity to photoperiod even after diapause, others lose it and become photo-refractory (temporarily or permanently). In Pentatomidae, such seasonal adaptations as photoperiodic control of nymphal growth rates, seasonal body colour change, migrations, and summer diapause (aestivation) are widely represented, whereas wing and/or wing muscle polymorphism has not been reported yet. In the subfamily Podopinae, induction of facultative reproductive winter diapause is under the control of photoperiod and temperature. All species feed on seeds and their seasonal development to a great extent reflects availability of food. However, the same food preferences and pattern of seasonal development are also characteristic to many species from the subfamily Pentatominae. All species of the subfamily Asopinae are predators. Among them, Picromerus bidens and Apateticus cynicus have obligate embryonic winter diapause, which is rear among true bugs. At the same time, A. cynicus and Podisus maculiventris belong to the same tribe but have different types of diapause: obligate embryonic diapause in A. cynicus and facultative adult diapause in P. maculiventris. Other Asopinae species studied up to date have facultative adult diapause controlled by photoperiod and temperature with probably only one exception: in Andrallus spinidens, adult diapause is controlled by temperature, and photoperiod plays only a secondary role. Thus, in spite of the similar habits and feeding types among Asopinae, the species of this subfamily have different types of diapause and the latter is controlled by different factors. In the subfamily Pentatominae, most species overwinter as adults and induction of their diapause is controlled by the long-day type photoperiodic response, in spite of the differences in their feeding preferences (within phytophagy). However, there are some exceptions in this subfamily, too: Palomena prasina, P. angulosa and Menida scotti have obligate diapause, which conditions univoltinism in these pentatomids. In M. scotti, only females have obligate adult diapause, whereas males remain physiologically active through the whole winter, this pattern being unusual for Heteroptera. The univoltine seasonal cycle of this species with summer diapause (aestivation) and apparent migrations is similar to that of shield bugs (Scutelleridae). According to the analysis of seasonal development, the evolution of seasonal adaptations in Pentatomidae does not directly reflect their phylogeny. However, individual genera, small tribes or even subfamilies have similar complexes of seasonal adaptations. At the same time, Pentatominae is a large and apparently collected taxon, but most of species in this subfamily have the same facultative adult diapause.     

Pythiopina,an enigmatic subtribe of darkling beetles (Coleoptera: Tenebrionidae: Pedinini): taxonomic revision,microtomography, ecological niche models and phylogenetic position

Morphological, anatomical, and distributional data concerning the S outh A frican endemic beetle subtribe P ythiopina (T enebrionidae: P edinini) are revised. Five species, representing two genera, are recognized. Included in this total is one new species (Meglyphus mariae K amiński sp.n. ). The following species are placed in synonymy: Meglyphus ciliatipes [=Meglyphus calitzensis syn.n. ]; Meglyphus laenoides [=Meglyphus andreaei syn.n. ; =Meglyphus namaqua syn.n. ]. Microtomographic models for all valid P ythiopina species, including the holotype of the newly described species, are presented and analysed. Endoskeleton morphology (specifically characters of the tentorium and metendosternite) proved to be informative at the specific and generic levels. An identification key is provided to all known species of the subtribe. Environmental niche models are presented for the majority of species. A molecular phylogeny of P edinini based on six genetic loci (28S : D 1–D 3 region; 28S : D 4–D 5 region, COII , A rgK , CAD 2, wg) was also produced to explore the phylogenetic position of P ythiopina. This analysis is the first to include representatives of all seven subtribes of P edinini, and supports a sister relationship between P ythiopina and the P alaearctic subtribe D endarina. Results also suggest the existence of a second pair of sister taxa within P edinini (in addition to M elambiina) with an amphitropical A frican distribution. This published work has been registered in ZooBank, http://zoobank.org/urn:lsid:zoobank.org:pub:285AD87A‐46B1‐4FE9‐BC57‐949EA1F70D49 .     

Structure of ovaries of scale insects: ii. margarodidae (INSECTA,hemiptera, COCCINEA)

The paired, spindle-shaped ovaries of the second instar of the Polish cochineal, Porphyrophora polonica (L.) (Hemiptera: Coccinea) are filled with cystocytes that are arranged into rosettes. In the centre of each rosette, there is a polyfusome. During the third instar, cystocytes differentiate into oocytes and trophocytes (nurse cells) and ovarioles are formed. Ovaries of adult females are composed of about 300 ovarioles of the telotrophic type. Each of them is subdivided into a tropharium (trophic chamber) and vitellarium. The tropharium consists of trophocytes and arrested oocytes that may develop. The number of germ cells in the trophic chambers varies from 11 to 18 even between the ovarioles of the same ovary. The obtained results seem to confirm the concept of a monophyletic origin of the primitive scale insects (Archaeococcoidea).