Genetic differentiation and distribution routes of the bronze featherback Notopterus notopterus (Osteoglossiformes: Notopteridae) in Indochina

Mitochondrial DNA analysis was used to investigate genetic variation in the bronze featherback fish Notopterus notopterus in Indochina. Five hundred and five fish samples collected from 11 localities were separated into three genetically differentiated intraspecific groups. Chao Phraya River samples formed a distinct group. Mekong River samples were clearly separated into two different groups, namely, middle and lower Mekong groups. The lower Mekong group was closely related to Malay Peninsula group, even though the two groups are separated by a tract of ocean. F‐values (0.82–0.95) also confirmed genetic differentiation of the middle and lower Mekong groups. Although the two Mekong groups inhabit the same river basin, molecular clock calculations indicate they separated approximately 1.2 Mya, suggesting (1) evolutionary divergence before invasion of the Mekong River; (2) differences in migration routes to the river; and/or (3) differences in the timings of invasion. The complexities of historical events cannot be ignored in determinations of factors responsible for the exceptional biodiversity in the Mekong basin. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 575–582.     

The Effect of Benzyladenine on Changes in Nuclease and Protease Activities in Intact Bean Leaves during Ageing

Primary leaves of intact bean plants (Phaseolus vulgaris L.) were treated with benzyladenine (BA) at different stages during growth. Changes in DNase, RNase, and proteas activities in the leaves were followed. Unlike the case of various excised tissues, cytokinin raised the activities of these hydrolases in intact bean leaves. Because BA elevated the levels of DNA, RNA, and protein in intact leaves, it may stimulate both synthesis and decomposition of these cellular constituents. The hydrolase activities showed differential responses to BA according to the age at which the leaf received the hormone treatment.     

Ion gradients in xylem exudate and guttation fluid related to tissue ion levels along primary leaves of barley

The concentration of ions in plant cells and tissues is an essential factor in determining physiological function. In the present study, we established that concentration gradients of mobile ions exist in both xylem exudates and tissues within a barley (Hordeum vulgare) primary leaf. For K⁺ and NO₃^?, ion concentrations generally decreased from the leaf base to the tip in both xylem exudates and tissues. Ion gradients were also found for Pi and Cl^? in the xylem. The hydathode strongly absorbed Pi and re‐translocated it to the rest of the plant, whereas Cl^? was extruded. The ion concentration gradients developed early during leaf growth, increased as the tissue aged and remained under both high and low transpiration conditions. Measurement of the expression profiles of Pi, K⁺ and NO₃^? transporters along the longitudinal axis of the leaf revealed that some transporters are more expressed at the hydathode, but for most transporters, there was no significant variation along the leaf. The mechanisms by which longitudinal ion gradients develop in leaves and their physiological functions are discussed.     

Determinants of feeding performance in free-ranging pit-vipers (Viperidae: Ovophis okinavensis): key roles for head size and body temperature

Feeding performance (handling time, capture success) in numerous animal species is well known to be influenced by a variety of ecological, functional, and physiological factors. Nonetheless, few studies have tested which factors are the strongest determinants of animal feeding performance in the wild. Using a field-based experiment, we examined the relationships among a number of functionally important variables and the predatory behaviour of free-ranging pit-vipers ( Ovophis okinavensis ) from Okinawa Island, Japan. Our main findings were: (1) strike latency was negatively related to snake body temperature and, hence, hotter snakes struck at frogs more readily than colder snakes; (2) initial bite position was correlated with ingestion direction (headfirst versus hindfirst) but ingestion direction was not correlated with ingestion duration; and (3) both snake head length and body temperature were negatively related with ingestion duration and, thus, snakes with longer heads and higher body temperatures had shorter ingestion durations. In O. okinavensis , head size and body temperature are therefore likely to have direct ecological consequences in terms of its feeding rate on explosively breeding frogs. More generally, however, this field-based study adds to the growing body of literature demonstrating that temperature has a pervasive influence on the feeding performance of ectotherms in general.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 93 , 53–62.     

The ontogeny of the platycopid Keijcyoidea infralittoralis (Ostracoda: Podocopa)

Platycopid ostracods such as the genus Keijcyoidea Malz, 1981 (Family Cytherellidae) have a unique body plan and are regarded as a phylogenetically ancient lineage. Their ontogeny, which is important in considering phylogenetical relationships, is very poorly known except for the growth of the carapace; there are nine instars including the adult, as in other podocopan groups. All appendages in all immature instars (A-8 to A-1) of Keijcyoidea infralittoralis Tsukagoshi, Okada & Horne, 2006 are described and illustrated here. The anlagen of the copulatory organs and the sexual dimorphism of carapace size appear in the sixth (A-3) instar, whereas sexual dimorphism in both the fifth and the sixth limbs, a distinctive feature of adults, is not clearly evident until the eighth (A-1) instar. Appendages are added at the moults between the second (A-7) and third (A-6), and between the fifth (A-4) and sixth (A-3) instars. The seventh limb, which platycopid ostracods have lost in the adult stage, is observed as an anlage in the sixth (A-3) and seventh (A-2) instars. During the other moults, there are no significant changes to the body plan. The ontogeny of the Platycopida is compared with that of the Podocopida, and strongly suggests that the phylogenetic position of the Platycopida is as an end-member of the Podocopa. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society , 2008, 153 , 213–237.     

Chemical Analysis of Melanin Pigments in Feather Germs of Japanese Quail Bh (Black at Hatch) Mutants

Bh (black at hatch) is a mutation of Japanese quails which causes darkening or lightening of the plumage in heterozygotes or homozygotes, respectively. We chemically analyzed melanin pigments in feather germs of Bh mutant embryos and in feathers of adult animals. Dark brown dorsal feathers of wild-type adult animals had white barrings, but heterozygous ones lacked clear barrings. The feathers of wild-type and heterozygote animals contained both eumelanins and pheomelanins, the latter being more pheomelanic. On the dorsal skin of 10-day old wild-type embryos, longitudinal stripes from black and yellow rows of feather germs developed; two or three longitudinal rows of black feather germs and then two or three rows of yellow feather germs next to the short central feather germs. Heterozygous embryos appeared black in plumage pigmentation, due to the presence of‘gray’feather germs in rows of dorsal feather germs that corresponded to yellow rows in wild-type embryos. Homozygous dorsal feather germs did not develop the black and yellow longitudinal stripes, but were brown. Chemical analysis showed that embryos of each genotype contained both eumelanins and pheomelanins in the feather germs; however, the eumelanin content in ho-mozygous feather germs was very low. These results suggest that the Bh mutation causes pheomelanic changes in feathers of quails.     

Regulatory mechanisms in phenotypic plasticity of diapause and nondiapause pupal colouration of the swallowtail butterfly Papilio machaon

Nondiapause pupae of Papilio machaon L. exhibit pupal colour diphenism comprising green–yellow and brown–white types. To understand the regulatory mechanism underlying the control of pupal colouration in P. machaon, the effect of environmental cues on diapause and nondiapause pupal colouration is investigated. When larvae reared under short‐day and long‐day conditions are allowed to pupate in sites with a smooth surface and a yellow background colour, all diapause pupae exhibit a brown–white type and 89.5% of nondiapause pupae exhibit a green–yellow type, respectively. With rough‐surface pupation sites, all diapause pupae exhibit brown–white and intermediate types, whereas a large proportion of nondiapause pupae exhibit brown–white and intermediate types, although some exhibit a green–yellow type. When extracts prepared from the head‐thoracic and thoracic‐abdominal regions of larval central nervous systems are injected into the ligated abdomens of P. machaon short‐day pharate pupae, all recipients exhibit a brown–white colouration. Furthermore, when each extract is injected into the ligated abdomen of Papilio xuthus L. short‐day pharate pupae with orange‐pupa‐inducing factor activity, recipients injected with the head‐thoracic extract exhibit the brown type, whereas those injected with the thoracic‐abdominal extract exhibit an orange colour. The results indicate that the response to the environmental cues of pupation site in P. machaon changes according to the photoperiodic conditions experienced during larval stages, and that at least two hormonal factors producing brown–white pupae are located in the larval central nervous system, with the secretion of these factors being regulated by the recognition of environmental cues in long‐day larvae.     

Species Specificity of the Insect Prothoracicotropic Hormone (PITH): the Presence of Bombyx-and Samia-Specific PTTHs in the Brain of Bombyx mori

Crude extracts of Bombyx mori brains can provoke adult development when injected into brain-removed dormant pupae of Bombyx mori and Samia Cynthia ricini. From this fact the prothoracicotropic hormone (PTTH) of Bombyx has long been thought to be species-nonspecifically active on Samia. Chemical fractionation of Bombyx brain or head extracts by fractional precipitation with acetone, Sephadex G-50 gel-filtration, and DEAE-Sepharose CL-6B chromatography, however, separated the fractions which activated Bombyx brainless pupae from those which activated Samia. Those results reveal the existence of two species-specific PTTHs.     

Effects of Thioureas, Inhibitors of Melanogenesis, on Lens Transdifferentiation of Cultured Chicken Embryonic Retinal Pigment Cells

The effects of phenylthiourea (PTU) and its analogues on chick embryonic pigmented epithelial cells (PECs) in culture were studied to elucidate the correlation between inhibition of melanogenesis of PECs and enhancement of their transdifferentiation into lens cells.
Both 0.25–0.5 mM PTU and 0.1 mM alpha-naphthylthiourea (ANTU) effectively inhibited melanogenesis of PECs and stimulated their transdifferentiation into lens cells at the same time. Thiourea (TU) also inhibited melanogenesis at a much higher concentration (4 mM), but did not stimulate the lens transdifferentiation at all. Methylthiourea (MTU), on the other hand, did not inhibit melanogenesis, but stimulated the lens transdifferentiation. Testicular hyaluronidase effectively amplified the above-mentioned stimulating effects of thioureas without their altering optimum concentrations, although this enzyme itself never enhanced the lens transdifferentiation of PECs but suppressed their melanogenesis at a concentration of 100 U/ml medium, onward.
These results suggest that the suppression of melanogenesis of PECs by PTU or its analogues does not directly correlate with their transdifferentiation into lens cells. The possible mode of thiourea actions on the lens transdifferentiation of PECs cultured in vitro is discussed.     

Quantitative Analysis of Protein Synthesis Altered by Estrogen in Cultured Xenopus Liver Parenchymal Cell

Using the primary culture system of male Xenopus laevis hepatocytes consisting of more than 95% parenchymal cells, the effect of estradiol-17 β (10⁻⁶M) on protein synthesis was quantitatively analyzed by ³H-leucine incorporation kinetics and the estimation of specific radioactivity of newly synthesized secretory protein. The cells in a well defined culture revealed high plating efficiency and very low DNA synthetic activity. The cultured cells could synthesize several secretory proteins containing serum albumin. The pattern of secreted proteins in SDS-polyacrylamide gel electrophoresis (SDS-PAGE) did not alter with culture time but secretion rate of protein increased for 7 days, starting on the third day following inoculation. Estradiol added to the culture media extensively induced the synthesis of yolk precursor protein vitellogenin which accounted for 40–50% of the overall secretory protein synthesis and 20–30% of the total protein synthesis on day 7 of estradiol treatment. Ultimately, the total protein synthesis and secretory protein synthesis were stimulated 1.2–1.3 fold and 2.0–2.2 fold, respectively, over those of the control cells cultured in the absence of estradiol. These results indicated that the stimulation of protein synthesis was largely due to vitellogenin production. Such an estradiol-dependent stimulation of protein synthesis was also detected in the low molecular weight protein(s). On the other hand, albumin synthesis was evidently reduced by estradiol. Thus, estradiol had two different effects on protein synthesis.
The results obtained in this study will be discussed in relation to the findings o in vivo experiments.