Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

The objective was to develop an efficient protocol for cryopreservation of agouti (Dasyprocta aguti) ovarian tissue. Agouti ovarian fragments were placed, for 10 min, in a solution containing MEM and fetal bovine serum plus 1.5 M dimethyl sulfoxide (DMSO), ethylene glycol (EG) or propanediol (PROH); some of those fragments were subsequently cryopreserved in a programmable freezer. After exposure and/or thawing, all samples were fixed in Carnoy prior to histological analysis. To evaluate ultrastructure, follicles from the control and all cryopreserved treatments were fixed in Karnovsky and processed for transmission electron microscopy. After exposure and freezing, there was a significant decrease in the percentage of morphologically normal preantral follicles in all treatments when compared to the control (92.67 ± 2.79, mean ± SD). However, there were no significant difference when the exposure and freezing procedures were compared using the same cryoprotectant. Moreover, there was no significant difference among cryoprotectants at the time of exposure (DMSO: 64.7 ± 3.8; EG: 70.7 ± 11.2, PROH: 63.3 ± 8.5) or after freezing (DMSO: 60.6 ± 3.6, EG: 64.0 ± 11.9; PROH: 62.0 ± 6.9). However, only follicles frozen with PROH had normal ultrastructure. In conclusion, preantral follicles enclosed in agouti ovarian tissue were successfully cryopreserved using 1.5 M PROH, with satisfactory maintenance of follicle morphology and ultrastructure.     

Development,morphology and ultrastructure of the branchial crown of Fabricia stellaris (Müller, 1774) (Polychaeta: Sabellida: Fabriciinae)

Randel, N. and Bick, A. 2011. Development, morphology and ultrastructure of the branchial crown of Fabricia stellaris (Müller, 1774) (Polychaeta: Sabellida: Fabriciinae). —Acta Zoologica (Stockholm) 93 : 409–421. Sabellidae and Serpulidae are well‐known tube‐building polychaetes with a distinctive and often spectacularly colourful branchial crown. Morphological investigations suggest that these taxa form the monophyletic clade Sabellida, with the adelphotaxa Sabellidae and Serpulidae, but the relationship between these taxa remains ambiguous. Molecular investigations have indicated that the Fabriciinae, major taxon of Sabellidae, belongs to Serpulidae, thereby making Sabellidae paraphyletic; however, morphological characters are absent to support this result. We investigate the development, anatomy and ultrastructure of the branchial crown of Fabricia stellaris (Müller, 1774), describing morphological characteristics useful not only for constructing morphological phylogenies but also for understanding the evolution of the branchial crown. The morphology of the radioles and pinnules does not differ from each other. The supporting tissue of the branchial crown consists of myoepithelial cells and a solid extracellular matrix (ECM). Both ciliated and non‐ciliated cells form the epidermal layer; ciliated cells shape the food groove. Most important is the result that radioles and pinnules within Sabellida may not be homologous, because the morphology and the branching of radioles and pinnules are completely different between Sabellinae, Fabriciinae and Serpulidae. The terms ‘primary branch’ for radioles and ‘secondary branch’ for pinnules are proposed for Fabriciinae. The phylogeny of the Sabellida is discussed.     

The male postabdomen and reproductive system of Bibio marci Linnaeus, 1758 (Hexapoda: Diptera: Bibionidae)

The male postabdomen and the internal parts of the male genital system of Bibio marci (Bibionomorpha) were examined and reconstructed 3‐dimensionally. Several features differ from the presumptive dipteran groundplan. The bases of the gonopods are fused with each other and with tergite IX. The penis is not tube‐shaped and only sclerotized on the ventral side. The vasa deferentia are S‐shaped, and two pairs of accessory glands are present. In contrast to these characteristics, the arrangement of the internal parts is probably close to the ancestral condition. With its specific shape, the penis is well suited for the transfer of a spermatophore. The dorsal sclerite of the copulatory organ probably represents the medially fused parameres. A cladistic analysis of 27 characters of the postabdomen yielded two most parsimonious trees, with the strict consensus as follows: Nannochoristidae (outgroup) + (Culicidae [Culicomorpha] + ((Nymphomyiidae + (Tipulidae + Trichoceridae)) + (Tabanidae [Brachycera] + (Bibionidae, Anisopodidae, Axymyiidae [Bibionomorpha])))). Potential synapomorphies of Bibionomorpha (including Axymyiidae) and Brachycera are the fusion of sternum IX with the gonocoxites, the fusion of the parameres forming the dorsal sclerite and the presence of an entire series of postabdominal muscles (M4, M20, M23, M26, M27, M31, M35 and M37). The results of the analysis are preliminary as it is based on a single‐character system with a limited taxon sampling. However, the main result – a clade Bibionomorpha + Brachycera – is fully compatible with current hypotheses on dipteran phylogeny.     

An integrative taxonomic revision of the Tarentola geckos (Squamata,Phyllodactylidae) of the Cape Verde Islands

Recent phylogeographical analyses using mitochondrial DNA (mtDNA) sequences indicate that the Tarentola geckos from the Cape Verde archipelago originated from a propagule that dispersed from the Canary Islands approximately 7.7 Mya and that underwent a fast evolutionary radiation. Molecular analyses carried out to date clearly show some incongruences with the current taxonomy of Tarentola from the Cape Verde Islands, with some species being paraphyletic or polyphyletic, and several independently evolving lineages needing formal taxonomic recognition. The aim of this study was to clarify the systematics of this group to unravel its taxonomy by applying an integrative approach based on information from three independent sources: mtDNA, nuclear genes, and morphology. As a result of this taxonomic revision, two novel species for the islands of S. Nicolau and Fogo are described and eight subspecies are upgraded to species level. Moreover, an identification key for the genus Tarentola from the Cape Verde archipelago is presented. This study reconciles taxonomy and phylogeny in this group, provides a better understanding of diversity patterns, new insights on evolutionary hypotheses, and supports the basic framework for the future management and conservation of this unique reptile radiation. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 328–360.     

Description and phylogeny of a new Neotropical genus of Acalyptini (Coleoptera: Curculionidae: Curculioninae) associated with the staminodes of Cyclanthaceae

The weevil genus Azotoctla gen. nov. (Coleoptera: Curculionidae: Curculioninae: Acalyptini Thomson), is described to accommodate the following 15 new Neotropical species: Azotoctla aecuatorialis sp. nov. (Ecuador), Azotoctla anerunca sp. nov. (Ecuador), Azotoctla angustacra sp. nov. (Ecuador), Azotoctla curvirostra sp. nov. (Peru), Azotoctla dasygastra sp. nov. (Costa Rica and Honduras), Azotoctla femorata sp. nov. (Colombia, Costa Rica, Ecuador and Panama), Azotoctla gomezi sp. nov. (Colombia, Costa Rica and Panama), Azotoctla gottsbergeri sp. nov. (Brazil), Azotoctla clemmyssa sp. nov. (Colombia), Azotoctla melolauta sp. nov. (Colombia), Azotoctla migueli sp. nov. (Colombia), Azotoctla nana sp. nov. (Panama), Azotoctla punctata sp. nov. (Costa Rica), Azotoctla tibiatra sp. nov. (Belize), and Azotoctla tuberquiai sp. nov. (Colombia). Azotoctla is placed in the tribe Acalyptini, subtribe Staminodeina Franz, thus representing the sister taxon of Staminodeus Franz. The monophyly of Azotoctla is supported by the following inferred traits: a noncarinate rostrum that is tumescent above the antennal insertion in males; equilaterally subtriangular hemisternites of the male sternum 8; paired, longitudinal, laterally positioned sclerites of the aedeagus; and a bifurcate lamina of the female sternum 8, which bears explanate, diverging furcal arms. A key to the species of Azotoctla is provided; and the salient features of each species are illustrated and presented along with distributional and natural history data. A morphological cladistic analysis of 23 terminal species (eight outgroup, 15 ingroup) and 33 characters yielded a single most parsimonious cladogram (length = 75 steps, consistency index = 52, retention index = 77), with the ingroup topology ((A. aecuatorialis, (A. anerunca, A. gottsbergeri)), (A. punctata, ((A. angustacra, A. nana), (A. tibiatra, (A. clemmyssa, (A. curvirostra, A. femorata)), (A. melolauta, ((A. gomezi, A. migueli), (A. dasygastra, A. tuberquiai))))))). Species of Azotoctla are reproductively associated with the ephemeral staminodes of different members of the Neotropical monocot family Cyclanthaceae; most commonly with the widespread genus Carludovica Ruiz & Pavón. The interspecific homogeneity in external morphology, coupled with marked differences in genital traits and an apparent absence of narrow geographical ranges mediated by either host plants and/or biogeographical factors, jointly suggest that the diversification of Azotoctla is a relatively recent phenomenon and driven strongly by sexual selection. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 166 , 559–623.     

Adaptations for nectar‐feeding in the mouthparts of long‐proboscid flies (Nemestrinidae: Prosoeca)

The insects with the longest proboscis in relation to body length are the nectar‐feeding Nemestrinidae. These flies represent important pollinators of the South African flora and feature adaptations to particularly long‐tubed flowers. The present study examined the morphology of the extremely long and slender mouthparts of Nemestrinidae for the first time. The heavily sclerotized tubular proboscis of flies from the genus Prosoeca is highly variable in length. It measures 20–47 mm in length and may exceed double the body length in some individuals. Proximally, the proboscis consists of the labrum–epipharynx unit, the laciniae, the hypopharynx, and the labium. The distal half is composed of the prementum of the labium, which solely forms the food tube. In adaptation to long‐tubed and narrow flowers, the prementum is extremely elongated, bearing the short apical labella that appear only to be able to spread apart slightly during nectar uptake. Moving the proboscis from resting position under the body to a vertical feeding position is accomplished in particular by the movements of the laciniae, which function as a lever arm. Comparisons with the mouthparts of other flower visiting flies provide insights into adaptations to nectar‐feeding from long‐tubed flowers. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, ?? , ??–??.     

Comparative floral structure and systematics in Ochnaceae s.l. (Ochnaceae,Quiinaceae and Medusagynaceae; Malpighiales)

Ochnaceae s.l. (Ochnaceae, Quiinaceae and Medusagynaceae), one of the well‐supported subclades of the large order Malpighiales retrieved so far in molecular phylogenetic studies, were comparatively studied with regard to floral structure using microtome section series and scanning electron microscopy (SEM). Floral morphology, anatomy and histology also strongly reflect this close relationship. Potential synapomorphies of the subclade include: flowers nectarless, sepals of different sizes within a flower, petals not retarded in development and forming the protective organs of advanced floral buds, petal aestivation contort, petals with three vascular traces, petals reflexed over the sepals and directed toward the pedicel, polystemony, anthers almost or completely basifixed, gynoecium often with more than five carpels, short gynophore present, styles separate for at least their uppermost part and radiating outwards, suction‐cup‐shaped stigmas, vasculature forming a dorsal band of bundles in the upper stylar region, gynoecium epidermis with large, radially elongate cells, ovules either weakly crassinucellar or incompletely tenuinucellar with an endothelium, abundance of tanniferous tissues and sclerenchyma in floral organs. The most strongly supported subclade of two of the three families in molecular analyses, Quiinaceae and Medusagynaceae, is also particularly well supported by floral structural features, including the presence of functionally and morphologically unisexual flowers, a massive thecal septum that persists after anther dehiscence, styles radiating outward from the ovary, two lateral ovules per carpel, positioned one above the other, conspicuous longitudinal ribs on the ovary wall at anthesis, and a ‘false endothelium’ on the nucellus at anthesis. Additionally, the group fits well in Malpighiales and further emphasizes the relationship of Malpighiales with Celastrales and Oxalidales, and thus the unity of the COM clade. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 170 , 299–392.     

Floral synorganization and its influence on mechanical isolation and autogamy in Marantaceae

The flowers of Marantaceae (～ 550 species) exhibit a highly derived pollination mechanism within Zingiberales, with a rapid and irreversible style movement based on a close synorganization of different floral parts. Given the complexity of the structure, we assume that little variation is possible if functionality is to be maintained. To test this, we investigated how much floral diversity exists in the clade and whether this diversity potentially influences the breeding system and placement of pollen on the pollinator. Flowers of 66 species covering the five major phylogenetic clades of the family were analysed. All species are similar in their basic flower construction: the fleshy staminode forms the tunnel‐shaped roof of the flower and narrows the tube with stiff swellings, and the hooded staminode holds the style under tension and narrows the flower entrance with its trigger appendage. Despite morphological diversity of the pollination apparatus, functionality is maintained by coordinated variation of the fleshy and hooded staminodes. Autogamy is usually avoided by herkogamy. However, in a few exceptions, subtle morphological changes alter the breeding system from allogamy to autogamy. Variable floral proportions allow for differential pollen deposition potentially causing mechanical isolation between sister taxa. This study clearly illustrates that structural variation is not only present in the highly synorganized flowers of Marantaceae, but that it also creates potentially new options for evolutionary diversification. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 168 , 300–322.     

Intraspecific versus interspecific variation in Miocene Great Basin mylagaulids: implications for systematics and evolutionary history

The Mylagaulidae are a family of burrowing rodents abundant in Miocene faunas from western North America. Recent taxonomic revisions of mylagaulids from the Great Plains suggest that their systematics may be best understood on a regional basis. Previous studies addressed the taxonomy and evolutionary history of mylagaulids from the Great Basin, but recent discoveries of specimens, new phylogenetic data, and more detailed stratigraphical information necessitate a thorough reanalysis of their relationships and occurrences. We present a revision of the systematics of the mylagaulids from the Great Basin. In addition to rare large mylagaulids of uncertain taxonomic affinity, we recognize four species of mylagaulids distributed throughout Oregon and Nevada from the late Hemingfordian through to the early late Hemphillian: Alphagaulus vetus, Hesperogaulus gazini, Hesperogaulus wilsoni, and a new species from the genus Hesperogaulus. All species are known from large sample sizes of isolated premolars, allowing consideration of ontogenetic variation in determining the key morphological differences that allow recognition of different species. Although the number of enamel lakes varies within a given taxon, the presence of some of these lakes is taxonomically significant. This result emphasizes the importance of understanding ontogeny in describing species of fossil hypsodont mammals. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 427–450.