Reconstitution of Embryo-Like Structures from Sea Urchin Embryo Cells: Distinction Between Cell-Cell and Cell-Substrate Associations

Sea urchin embryos can be dissociated into a suspension of single cells that reconstitute embryo-like structures. When reconstitution is conducted in stationary cultures the first step is attachment of the cells to the culture plate, which requires calcium and metabolic energy but not protein synthesis. We have found that protease treated cells form cell-cell associations in stationary cultures without attaching to the culture plates, and that cell-plate attachments are unaffected by inhibition of protein synthesis. These data suggest that cell surface proteins are needed for cell-plate attachment and that these proteins are present on freshly dissociated cells. We also demonstrated that butanol extracted cells attach to the plates, but do not form functional cell-cell associations unless the butanol extracted material is restored to them. We conclude that sea urchin embryo cells contain two classes of attachment components. The first class functions in the cell-plate attachments, is protease sensitive, and not extracted by butanol; the second class is necessary for cell-cell associations, is protease insensitive, and extracted by butanol. Since protease treated cells reconstitute embryo-like structures without attaching to the culture plates, only the second class of attachment components is necessary for embryo reconstitution.     

Adaptive evolution of cryptic coloration: the shape of host plants and dorsal stripes in Timema walking-sticks

The adaptive significance of cryptic colour patterns has seldom been analysed in a phylogenetic context. We mapped data on the presence vs. absence of dorsal stripes, and the use of needle-like vs. broad foliage, onto a recent phylogeny of Timema walking-sticks, in order to infer the evolutionary history of these traits and test the hypothesis that the dorsal stripe is an adaptation for crypsis on needle-like leaves. By maximum parsimony optimization, the dorsal stripe has evolved five or six times in this clade, each time in association with the use of vegetation with needle-like leaves. Maddison's concentrated changes test showed that this association between morphology and habitat was statistically significant. By contrast, results based on Pagel's maximum likelihood analyses did not reach significance, probably because the large number of origins of dorsal stripe introduces statistical uncertainty. These results suggest that the adaptations for crypsis can arise readily and in parallel, in the appropriate selective environment. However, they may also constrain the evolution of host-plant use, as there is no unambiguous case of Timema species with dorsal stripes shifting to broad-leaved plants.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 1–5.     

Isolation of microsatellite markers from Pinus densiflora Sieb. et Zucc. using a dual PCR technique

We developed seven microsatellite loci from Pinus densiflora using a dual polymerase chain reaction (PCR) technique. Of 186 clones from a library based on suppression PCR, 127 contained microsatellite sequences. Of these, 43 candidates were determined sequences of both flanking regions, and 16 regions from this group were chosen as development markers. Seven of these primer pairs successfully amplified polymorphic single loci among 83 resistant trees against pine wood nematode. The observed heterozygosity of the seven microsatellite markers ranged from 0.247 to 0.843. Mendelian inheritance was confirmed using megagametophytes.     

Epidemiology of apple proliferation (AP) in northwestern Italy: evaluation of the frequency of AP-positive psyllids in naturally infected populations of Cacopsylla melanoneura (Homoptera: Psyllidae)

Adults of Cacopsylla melanoneura, vector of the apple proliferation (AP) phytoplasma, were collected every 2 weeks from January until May in 2000 and 2001 by the beating tray method in eight apple orchards of the Aosta Valley (northwestern Italy). Total DNA was extracted from batches of five insects and amplified with the universal phytoplasma primers P1/P7 in direct PCR. A nested PCR assay was then performed on P1/P7 amplicons using the primers fO1/rO1, specific for the AP‐ phytoplasma group. The digestion of fO1/rO1 amplicons with Ssp I restriction endonuclease confirmed that C. melanoneura adults harboured the AP phytoplasma. The data obtained with PCR were used to estimate the proportion of AP‐positive insects in over wintered and offspring adults. Percentages of AP‐positive insects of 3.6% and 0.8% were estimated in 2000 among over wintered and offspring psyllids respectively. In 2001 only the over wintered insects were found infected, with an estimated proportion of 2.8%. The seasonal abundance of the vector was measured using yellow sticky traps. C. melanoneura was always present at a low population level, and the highest density was recorded from mid‐February until mid‐March in both years. The results show that the overwintered population is higher and spends a longer period in apple orchards, suggesting the crucial role of the overwintered adults in vectoring AP.     

Isolation and characterization of 49 polymorphic microsatellite loci in the ayu,Plecoglossus altivelis

A total of 49 novel polymorphic microsatellite markers in the ayu fish, Plecoglossus altivelis, were successfully isolated and characterized. The number of alleles per locus, observed heterozygosity, and expected heterozygosity ranged from two to 25 (11.3 on average), from 0.125 to 1.000 (0.648 on average), and from 0.123 to 0.957 (0.751 on average), respectively. These microsatellite markers will be useful for both studying population structure and identifying quantitative trait loci that control evolutionarily and economically important traits of ayu.     

Paranucleospora theridion n. gen., n. sp. (Microsporidia,Enterocytozoonidae) with a Life Cycle in the Salmon Louse (Lepeophtheirus salmonis,Copepoda) and Atlantic Salmon (Salmo salar)

ABSTRACT. Paranucleospora theridion n. gen, n. sp., infecting both Atlantic salmon (Salmo salar) and its copepod parasite Lepeophtheirus salmonis is described. The microsporidian exhibits nuclei in diplokaryotic arrangement during all known life‐cycle stages in salmon, but only in the merogonal stages and early sporogonal stage in salmon lice. All developmental stages of P. theridion are in direct contact with the host cell cytoplasm or nucleoplasm. In salmon, two developmental cycles were observed, producing spores in the cytoplasm of phagocytes or epidermal cells (Cycle‐I) and in the nuclei of epidermal cells (Cycle‐II), respectively. Cycle‐I spores are small and thin walled with a short polar tube, and are believed to be autoinfective. The larger oval intranuclear Cycle‐II spores have a thick endospore and a longer polar tube, and are probably responsible for transmission from salmon to L. salmonis. Parasite development in the salmon louse occurs in several different cell types that may be extremely hypertrophied due to P. theridion proliferation. Diplokaryotic merogony precedes monokaryotic sporogony. The rounded spores produced are comparable to the intranuclear spores in the salmon in most aspects, and likely transmit the infection to salmon. Phylogenetic analysis of P. theridion partial rDNA sequences place the parasite in a position between Nucleospora salmonis and Enterocytozoon bieneusi. Based on characteristics of the morphology, unique development involving a vertebrate fish as well as a crustacean ectoparasite host, and the results of the phylogenetic analyses it is suggested that P. theridion should be given status as a new species in a new genus.     

Behavioral response of male mantid Tenodera aridifolia (Mantodea: Mantidae) to windy conditions as a female approach strategy

When sexual cannibalism presents a sexual conflict, one expects to find male traits that reduce the risk of cannibalism. In sexually cannibalistic species, selection is thought to have favored the evolution of male approach behaviors that reduce the probability that the female will kill the male. We tested the hypothesis that male mantids change their approach behavior in response to wind to reduce the risk of being noticed by females. Time between detection of the female by the male and mating was shorter under windy than windless conditions. Sexual approach behavior was observed more frequently under windy than windless conditions. Moreover, this behavior was observed more frequently when the female was walking than when the female was not walking under windy conditions. The detection rate of male mantids by females was significantly lower on swaying leaves than on fixed leaves. Our results thus indicate that male mantids were more active in response to wind. Therefore, we suggest that the male's quick approach strategy toward females when the wind is blowing at short range is adaptive in reducing the risk of detection by females.     

Pattern of conodont reworking in the Upper Devonian of the Alps: palaeoenvironmental and palaeotectonic implications

Transfer of sediments from shallow shelf to basin usually implies vertical (stratigraphic) reworking of fossils, but in a few instances produces only lateral reworking (actually within the time span of a subzone). An Upper Devonian (Frasnian) pelagic carbonate sequence with allodapic intercalations and thick intraclast parabreccias triggered by seismic shocks and related tsunami and turbidity currents has been studied. Here, the standard biozone sequence is undisturbed at the zonal as well as the subzonal level. However, distribution of conodonts is discontinuous, and ranges of some important species are altered; mature specimens are commonly broken, whereas complete specimens are mostly juvenile forms. Maxima and minima of element frequency and anomalous thickening or thinning of subzones are recognized. Such distribution patterns of conodonts are compared with the four main facies distinguished and related to the environmental interpretation (a platform-basin system, the original gentle slope of which was fragmenting in fault-controlled blocks). The mean frequency of shallow-water, proximal conodonts is 10% whereas transitional to distal deeper-water genera dominate throughout the entire section. In the dominant biofacies, conodonts settled vertically, in the subordinate one they were transported laterally. The 'intraclast parabreccia' is interpreted as a peculiar type of seismitc. The anomalous thickness variation of biozones is related to the step topography of the slope in a fault-controlled carbonate basin.