Phylogeny and divergence time of island tiger beetles of the genus Cylindera (Coleoptera: Cicindelidae) in East Asia

To examine the diverse colonization histories in eight tiger beetle species of the genus Cylindera (Coleoptera: Cicindelidae) on the East Asian islands, we conducted phylogenetic analyses and divergence time estimation using mitochondrial cytochome oxidase subunit I (COI) and nuclear 28S rDNA sequences. The island fauna consisted of four subgenera: Apterodela, Cicindina, Ifasina, and Cylindera. Apterodela is a flightless group with large bodies, whereas the others are fliers with small bodies. In Apterodela, the divergence among endemic species in Taiwan, Japan, and the mainland was ancient (2.1–4.7 Mya), as expected from their flightlessness. Their dispersal might have occurred across the extended landmass in East Asia during the Pliocene. In the subgenus Cicindina, Cylindera elisae has spread throughout East Asia, from which an endemic species, Cylindera bonina, was derived on the oceanic Bonin Islands during the early Pleistocene (0.9 Mya). This indicates the significance of Cylindera bonina, which is currently confined to a single island, for conservation. In the subgenus Ifasina, Cylindera kaleea is widely distributed in East Asia, and its sister species Cylindera humerula, endemic to Okinawa Island, diverged 1.0 Mya, whereas Cylindera psilica on Taiwan and the Yaeyama Islands diverged approximately 0.8 Mya. In the subgenus Cylindera, the colonization of Cylindera gracilis in Japan from the mainland occurred during the last glacial period. With the exception of C. bonina, which likely colonized new territories by flight or drifting, other dispersal events might have used land connections that occurred repeatedly during the Pliocene and Pleistocene. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2011, 102 , 715–727.     

Developmental morphology of the ovules of Amborella trichopoda (Amborellaceae) and Chloranthus serratus (Chloranthaceae)

The developmental morphology of the outer integument in the pendent orthotropous ovules of Amborella trichopoda (Amborellaceae) and Chloranthus serratus (Chloranthaceae) was studied. In both species the outer integument is semiannular at an early stage and becomes cup-shaped but dorsiventrally somewhat asymmetric at later stages. The outer integument, which is initiated first on the concave and lateral sides of the ovule, differs from that of the anatropous ovules of other basal families with the outer integument semiannular at an early stage or throughout development. The bilateral symmetry of the outer integument is shared by these orthotropous and anatropous ovules. The developmental pattern of the outer integument and ovule incurving characterize the ovule of the Amborellaceae and Chloranthaceae, which is not equivalent to typical orthotropous ovules of eudicots. A phylogenetic analysis of ovule characters in basal angiosperms suggests that anatropous ovules with cup-shaped outer integuments and orthotropous ovules were derived independently in several clades and that the ovules of Amborella and Chloranthus might also be derivative.     

Molecular phylogenetic analysis of Podostemaceae: implications for taxonomy of major groups

The river‐weed family Podostemaceae (c. 300 species in c. 54 genera) shows a number of morphological innovations to be adapted to its unusual aquatic habitat, and its unique or rare bauplan features have been reflected in the traditional (i.e. non‐molecular) classification recognizing numerous monotypic or oligospecific genera. The infrasubfamilial relationships of many genera remained unclear. The present study used molecular phylogenetic analysis of matK sequences for 657 samples (c. 132 species/c. 43 genera). The family was traditionally divided into three subfamilies (Podostemoideae, Tristichoideae and Weddellinoideae). American Podostemoideae were shown to be polyphyletic and divided into four clades, i.e. Ceratolacis, Diamantina, Podostemum and all other genera. Among the podostemoid clades, Diamantina was the first branching clade and a clade comprising Mourera and the Apinagia subclade was then sister to the remainder of the New World and Old World Podostemoideae with low statistic supports. The Old World Podostemoideae comprised four monophyletic clades, i.e. two African clades, one Madagascan clade and one Asian clade, although the relationships among these clades and American Ceratolacis and Podostemum were poorly resolved. African Podostemoideae were polyphyletic, with Saxicolella pro parte being weakly supported as sister to the remaining Old World Podostemoideae plus Ceratolacis and Podostemum. In contrast to the American and African clades, monophyly of four Asian subclades was well supported. Plants of Tristicha (Tristichoideae) and of Weddellina (Weddellinoideae), which are currently treated as monospecific, had great matK differentiation equivalent to at least interspecific variation. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169 , 461–492.     

Developmental Changes in the Activity of Xanthine Dehydrogenase and closely Related Enzyme in Chick Tissues

Xanthine-oxidizing activities in the chick tissues were measured by a couple of assay procedures during development of chick embryo. With the usual assay using pterine and NAD⁺, no detectable level of XDH activity was observed in the liver and little in the duodenum before hatching, whereas an appreciable activity was detected in the kidney of chick embryo. When assayed with xanthine and dichlorophenol indophenol, an XDH-like enzyme activity was significantly detected in the embryonic liver, while no further enhancement of the activity was detected in the kidney and duodenum. Electrophoregrams obtained with samples from various developmental stages, followed by activity staining with tetrazolium dye, supported the above results and revealed that the embryonic XDH-like enzyme is not distinguishable from XDH of adult tissue in molecular size. This XDH-like enzyme, pre-existing in the liver before hatching, however, exhibited no cross reaction with antibody against intact XDH. The nature of this material was discussed in comparison with deflavinated XDH.     

Detection of Mouse Tyrosinase With a Monoclonal Antibody MAT-1 Against Human Tyrosinase

In this study we explored the possible application of MAT-1, which has been established as a monoclonal antibody against human tyrosinase, for detection of mouse tyrosinase. The MAT-1 reacted with B16 mouse melanoma cells, but not with tyrosinase-negative NIH-3T3 mouse fibroblasts. In western blot analysis of the large granule fraction (LGF) of B16 cells, MAT-1 detected a single protein of 80 kDa, whose size was close to that of human tyrosinase detected with MAT-1 in extracts of human melanocytes. Furthermore, the 80 kDa band that was detected with MAT-1 in the LGF of B16 cells was also detected by DOPA reaction. In order to confirm that the protein detected with MAT-1 is tyrosinase, a transient expression assay was carried out. When mouse tyrosinase or mouse tyrosinase-related protein 1, which shares high homology with human tyrosinase, was transiently expressed in tyrosinase-negative K1735 mouse melanoma cells by cDNA transfection, MAT-1 reacted only with the cells expressing mouse tyrosinase. These results indicate that MAT-1 specifically reacts with mouse tyrosinase.     

Breeding system and floral visitors of Veratrum album subsp. oxysepalum (Melanthiaceae)

Veratrum album subsp. oxysepalum is an andromonoecious perennial woodland herb. In the present study, to clarify the essential life-history characteristics of this species, sexual reproduction was examined and breeding experiments were carried out along with observations of floral visitors in the field. The results of the breeding experiments showed that this species has a high level of self-incompatibility. As various Coleoptera and Diptera visited the flowers, the seed production of V. album subsp. oxysepalum possibly depends on outcrossing pollination by insects.     

Sexual dimorphism in Champsosaurus (Diapsida, Choristodera)

Two nearly complete specimens of Champsosaurus (Diapsida, Choristodera) with distinctive morphologies, from the Tullock Formation (Early Paleocene) of northeastern Montana, USA, were described as different species. The limb bones of C. ambulator are more robust than those of C. laramiensis, indicating that C. ambulator was more adapted for walking than C. laramiensis. The phylogenetic significance of these limb bone morphologies, however, appears questionable because similar dimorphic variations occur in a closely related genus and champsosaurs from other geologic ages and locations. Female champsosaurs may have been better adapted to a terrestrial life than males due to nesting behavior on land, resulting in variable limb bone morphologies between sexes. The observed morphologic variations are, hence, hypothesized to reflect sexual dimorphism rather than sympatry of species. The C. ambulator-shaped humeri and femora, demonstrating a terrestrial adaptation, are suggested to belong to females and C. laramiensis-shaped limb bones to males. No significant variations of humeral and femoral morphologies occur in small champsosaur specimens, suggesting an aquatic niche for juveniles like adult males.