Phylogenetic analysis of the Brazilian microendemic Paepalanthus subgenus Xeractis (Eriocaulaceae) inferred from morphology

Paepalanthus subgenus Xeractis (Eriocaulaceae) comprises 28 recognized species endemic to the Espinhaço Range, in Minas Gerais state, Brazil. Most species of the subgenus are restricted to small localities and critically endangered, but still in need of systematic study. The monophyly of the subgenus has already been tested, but only with a few species. Our study presents the first phylogenetic hypothesis within the group, based on morphology. A maximum parsimony analysis was conducted on a matrix of 30 characters for 30 terminal taxa, including all species of the subgenus and two outgroups. The biogeographical hypotheses for the subgenus were inferred based on dispersal–vicariance analysis (DIVA). The analysis provided one most‐parsimonious hypothesis that supports most of the latest published subdivisions (sections and series). However, some conflicts remain concerning the position of a few species and the relationships between sections. The distribution and origin(s) of microendemism are also discussed, providing the ground for conservation strategies to be developed in the region. © 2011 The Linnean Society of London, Botanical Journal of the Linnean Society, 2011, 167 , 137–152.     

Larval morphology and phylogenetic position of Horelophopsis hanseni Satô et Yoshitomi (Coleoptera,Hydrophilidae, Horelophopsinae)

The subfamily Horelophopsinae was originally proposed as one of the earliest diverging clades of Hydrophilidae (s.s.), but its phylogenetic placement has never been tested. We describe the larva of Horelophopsis hanseni Satô et Yoshitomi, 2004 of the Horelophopsinae. Larval data are based on larval specimens collected together with adults, and unambiguously associated with them by means of DNA barcoding. We perform an analysis testing the phylogenetic position of H. hanseni based on larval and adult morphological characters. Horelophopsis hanseni is unambiguously placed within the hydrophilid subfamily Hydrophilinae and its close relationships to the genus Agraphydrus Régimbart, 1903 (Hydrophilinae, Acidocerini) is recognized. The results suggest that the subfamily Horelophopsinae is unlikely to be a basal taxon of Hydrophilidae, as originally suggested.     

Female-biased sex ratio in a wild bruchid seed-predator, Kytorhinus sharpianus. I. Larval competition and other factors

Abstract.

• 1 A wild bruchid seed-predator, Kytorhinus sharpianus, has a complex life cycle consisting of bi- and trivoltinism on a wild leguminous plant, Sophola flavescens. Observations of adults showed significant female-biased sex ratios (from 1:2 to 1:6) for nine generations over 4 years.
• 2 To investigate the potential effects of larval competition on the sex ratio, we altered the number of hatched eggs per seed and counted emergent males and females under laboratory conditions. Although only one adult could emerge per seed, the ratio of the females that emerged increased with the number of hatched eggs per seed. However, the sex ratio was not significantly different from 1:1 in the case of one hatched egg per seed.
• 3 We dissected seeds bearing two hatched eggs at regular intervals, and classified the surviving and the dead larvae according to their developmental stage. Over time, one larva within each seed always survived, while the other larva died from the second to fourth instar before the seed resource became exhausted.
• 4 In order to study the effects of the difference in the stages of two larvae in a seed on the emergence sex ratio, we manipulated intervals between the first and second ovipositions in the laboratory. As the difference in developmental stages of the two larvae increased, the closer to 1:1 the emergence sex ratio became.
• 5 Field observations, however, showed that about 60% of infested seeds were bored by only one K.sharpianus larva. This suggests that female dominance in larval competition within a seed may be relatively unimportant in causing the female-biased sex ratio in the field.

     

Calcium- and Cyclic AMP-independent,Labile Protein Kinase Appearing during Starfish Oocyte Maturation: its Extraction and Partial Characterization*

A burst of protein phosphorylation and an appearance of maturation-promoting factor have been reported to occur shortly before germinal vesicle (nucleus) breakdown (GVBD) in 1-methyladenine-induced oocyte maturation of starfish. To detect if a protein kinase is activated before GVBD, protein kinase activity was compared in maturing oocytes which were just undergoing GVBD and immature oocytes of Asterina pectinifera. The oocytes were homogenized in a buffer modified from that used for extracting amphibian maturation-promoting factor. When the supernatant protein of homogenized immature oocytes was used as a substrate, protein kinase activity in the supernatant of the maturing oocytes was 7-fold higher than that of immature oocytes. The protein kinase in the supernatant of the maturing oocytes showed a high substrate specificity for histone H1 among the exogenous substrates examined, and the activity of the maturing oocytes for histone H1 was 6- to 7-fold higher than that of immature oocytes. The protein kinase detected in the maturing oocytes was very labile and was inhibited neither by ethylene glycol bis(β-aminoethyl ether)N, N, N′, N′-tetraacetic acid nor by the heat-stable inhibitor protein of cyclic AMP-dependent protein kinase. These results indicate that a calcium- and cyclic AMP-independent, labile “maturation-specific protein kinase” appeared before GVBD in maturing oocytes, and suggest its participation in the phosphorylation burst in vivo. The possible correlation of this kinase with maturation-promoting factor and chromosome condensation was discussed.     

DNA binding of citrus dehydrin promoted by zinc ion

Dehydrins are hydrophilic proteins that accumulate during embryogenesis and osmotic stress responses in plants. Here, we report an interaction between citrus dehydrin Citrus unshiu cold-regulated 15 kDa protein (CuCOR15) and DNA. Binding of CuCOR15 to DNA was detected by an electrophoretic mobility shift assay, a filter-binding assay and Southwestern blotting. The binding was stimulated by physiological concentrations of Zn²⁺, but little stimulation occurred when other divalent cations, such as Mg²⁺, Ca²⁺, Mn²⁺, Ni²⁺ and Cu²⁺, were substituted for Zn²⁺. Ethylenediaminetetraacetic acid cancelled the Zn²⁺-stimulated binding. A binding curve and competitor experiments suggested that the DNA binding of CuCOR15 exhibited low affinity and non-specificity. Moreover, tRNA competed with the DNA binding. Histidine-rich domains and a polylysine segment-containing domain participated in the DNA binding. These results suggest that CuCOR15 can interact with DNA, and also RNA, in the presence of Zn²⁺. Dehydrin may protect nucleic acids in plant cells during seed maturation and stress responses.     

Over-wintering stage polymorphism of a bruchine beetle: geographical variation in optimal diapause strategy

Abstract.  1. Most temperate insects undergo diapause at a specific developmental stage to overcome severe winters. The bruchine beetle Bruchidius dorsalis in a warmer region in Japan, however, has diverse over-wintering stages – non-diapausing larvae, diapausing larvae, and diapausing adults, whereas in a cooler region, it over-winters only as the final instar larva or adult in diapause.
2. The geographical pattern of over-wintering stages in 12 populations was investigated over a wide range along the mainland of Japan. It revealed that a distinct geographical pattern of over-wintering stages exists in relation to temperature conditions. In regions with warmer climates, the proportion of non-diapausing larvae increased and B. dorsalis had a more complex over-wintering stage structure.
3. Life cycles were also compared between two areas of Japan by field experiments. In the cooler area, the first generation over-wintered in the diapausing larval or adult stage. Conversely, in the warmer area, diapause was induced later and some of the first-generation adults produced second-generation offspring before over-wintering.
4. Based on the geographical cline of climates and the differences in cold hardiness among stages, we can demonstrate that the over-wintering stage variation among and within populations results from an adaptive timing of diapause induction in each region, because the late larval or adult diapauses protect pupae or eggs – which unlike other stages are not cold hardy – from being produced late in the season.     

ESTABLISHMENT OF A CELL LINE FROM EMBRYONIC TISSUES OF THE FLESHFLY, SARCOPHAGA PEREGRINA (INSECTA: DIPTERA)

A continuous cell line was established from embryonic tissues of the fleshfly, Sarcophaga peregrine , and was designated as NIH-SaPe-4. The primary culture was initiated in October, 1977, and the cell line was passed 68 times during the following year. The cells were heterogeneous in morphology. Most cells were diploid and their chromosomes consisted of 4 metacentric, 6 sub-metacentric and 2 micro chromosomes. The population doubling time of the cell line was about 30 hr. The cells grew faster in Mitsuhashi-Maramorosch's medium than in Schneider's medium. The cells were either stored in the usual medium at 5°C for about 3 months, or in a medium containing 10% glycerol at –80°C for a longer period. Cell growth was suppressed by 20-hydroxy-ecdysone when at a greater concentration than 0.01 μg/ml, whereas insulin showed no effects on cell growth at a strength of 0.4 and 0.04 IU/ml.     

First record of an Early Barremian caprinid rudist from Japan – implications for the palaeobiogeography of the Caprinidae (Bivalvia)

Abstract: Pachytraga Paquier, 1900, the stratigraphically oldest genus of caprinine caprinid rudist, was previously known from only two chronospecies from a single lineage, that is the Hauterivian P. tubiconcha Astre, 1961 and the early Aptian P. paradoxa (Pictet and Campiche, 1869). Here, a new species, Pachytraga? tanakahitoshii, is erected on the basis of isolated left valves recovered from the Osaka and Sanchu areas, south‐west Japan. This species has a moderate shell size (antero‐posterior commissural diameter c. 30 mm), and its left valve is characterized by at least one possibly autapomorphic character (narrow anterior myophore, inclined inwards), as well as a mosaic of primitive (single longitudinal carina developed on the anterior side) and derived (simple marginal canals in the antero‐dorsal valve margin) characters of Mediterranean and Middle East Tethyan Pachytraga. The Japanese Pachytraga? represents the first probable record of this genus outside the Mediterranean/Middle Eastern Tethyan province, and its early Barremian age partly fills the ‘gap’ in its previously known stratigraphical record, although the evolutionary relationship of the Japanese form with Mediterranean and Middle East Tethyan Pachytraga remains unsolved. However, the discovery of early Barremian Pachytraga? in Japan indicates that the evolutionary history of the genus is more complex than previously thought and should thus be discussed in a broader palaeogeographical context that must now include the Pacific.