Comparative anatomy and systematics of woody Saxifragaceae. Aphanopetalum Endl.

The floral and vegetative anatomy of the small Australian genus Aphanopetalum were studied. Wood is described for the first time and is characterized by predominantly solitary pores, scalariform vessel element perforation plates with low bar numbers, imperforate tracheary elements with distinctly bordered pits, sparse axial parenchyma, and a combination of homocellular and heterocellular rayS. Starch occurs in both axial and ray parenchyma of the wood. Stems possess unilacunar, one-trace nodes and the uncommon feature of an endodermis with well-defined Casparian stripS. Leaves have anomocytic stomata, a bifacial mesophyll and semicraspedodromous venation or a combination of semicraspedodromous and brochidodromous venation. The tetramerous flowers are apetalous or have minute petals. The compound, half-inferior gynoecium consists of essentially totally united carpels. The pattern of floral vascularization resembles different Saxifragaceae sensu lalo in that the compound sepal-plane and petal-plane traces give rise to staman bundles as well as sepal, petal, and carpel wall venation in their respective planes. The ventral ovarian bundles are fused into a single ventral complex that subdivides at the top of the ovary to form ventral bundles and to supply the one ovule in each locule. Vegetative and floral features provide compelling evidence to suggest that Aphanopetalum has its nearest relatives among the Saxifragaceae sensu lato rather than Cunoniaceae. The genus is probably best treated as forming its own subfamily (or family) among the saxifragaean alliance.     

Incubation in Mice Provides a Signal for the Differentiation of Trypanosoma cruzi Epimastigotes to Trypomastigotes1

The differentiation of Trypanosoma cruzi epimastigotes into trypomastigotes was studied in diffusion chambers sub-cutaneously implanted in mice. Using epimastigotes of the Tulahuén strain, transformation was first evident at 16 h after implantation and reached its maximum (92% trypomastigotes) by 24 h. Shortly before their differentiation into trypomastigotes, epimastigotes were found to develop resistance to lysis by the alternative pathway of complement. Furthermore, implantation of stationary-phase (as opposed to log-phase) parasites resulted in the accumulation of large numbers of complement-resistant epimastigotes in the chambers. These observations suggest that epimastigotes pass through a complement-resistant transitional stage before differentiating into trypomastigotes and that transformation may require cell division. In a further series of experiments, epimastigotes recovered 7 h after implantation in mice were found to differentiate into trypomastigotes when cultured in vitro for an additional 17 h at 37°C. This observation indicates that the events which trigger the morphologic transformation of epimastigotes into trypomastigotes can be dissociated operationally from the differentiation process itself.     

Polyamine Synthesis and Interconversion by the Microsporidian Encephalitozoon cuniculi

Polyamines are small cationic molecules necessary for growth and differentiation in all cells. Although mammalian cells have been studied extensively, particularly as targets of polyamine antagonists, i.e. antitumor agents, polyamine metabolism has also been studied as a potential drug target in microorganisms. Since little is known concerning polyamine metabolism in the microsporidia, we investigated it in Encephalitozoon cuniculi, a microspordian associated with disseminated infections in humans. Organisms were grown in RK-13 cells and harvested using Percoll gradients. Electron microscopy indicated that the fractions banding at 1.051-1.059/g/ml in a microgradient procedure, and 1.102-1.119/g/ml in a scaled-up procedure were nearly homogenous, consisting of pre-emergent (immature) spores which showed large arrays of ribosomes near polar filament coils. Intact purified pre-emergent spores incubated with [1H] ornithine and methionine synthesized putrescine, spermidine, and spermine, while [14C]spermine was converted to spermidine and putrescine. Polyamine production from ornithine was inhibitable by DL-alpha-difluoromethylornithine (DFMO) but not by DL-alpha-difluoromethylarginine (DFMA). Cell-free extracts from mature spores released into the growth media had ornithine decarboxylase (ODC), S-adenosylmethionine decarboxylase (AdoMetdc), and spermidine/spermine N1-acetyltransferase (SSAT) activities. ODC activity was inhibited by DFMO, but not by DFMA. AdoMetdc was putrescine-stimulated and inhibited by methylglyoxal-bis(guanylhydrazone); arginine decarboxylase activity could not be detected. It is apparent from these studies that Encephalitozoon cuniculi pre-emergent spores have a eukaryotic-type polyamine biosynthetic pathway and can interconvert exogenous polyamines. Pre-emergent spores were metabolically active with respect to polyamine synthesis and interconversion, while intact mature spores harvested from culture supernatants had little metabolic activity.     

Breeding biology during establishment of a reintroduced Griffon Vulture Gyps fulvus population

We studied the breeding parameters of a reintroduced population of individually marked Griffon Vultures Gyps fulvus in the Grand Causses region of southern France from the time of first reintroduction in 1982 to 1992. Among nesters, 65% of the birds released as immatures or born in the wild recruited into the breeding population when 4 years old, i.e. 1 year earlier than previously described. The proportion of birds nesting each year was relatively high and increased with time, suggesting that conspecific attraction favoured recruitment. We detected a permanent adverse effect of long-term captivity on the nesting success of birds released when more than 2 years old: compared to the natural population, birds which had been kept in captivity showed a reduced breeding success during the whole study period. The breeding success of released immatures and wild-born birds was similar to the highest values observed in a natural population in the Spanish and French Pyrenees. Breeding failures did not usually result in mate change but affected nest-site fidelity. Birds not born in the wild were more likely to recruit to the largest subcolonies available, which highlights the role of social attraction. The observed philopatry of wild-born birds probably resulted from such a social attraction since most of them were born in the largest subcolony. Conspecific attraction maintained the spatial aggregation of nests, whereas nest changes after a breeding failure favoured the spatial expansion of the colony.     

Characterization of polymorphic microsatellite markers for the endangered Malagasy radiated tortoise (Geochelone radiata)

The radiated tortoise (Geochelone radiata) is an endangered endemic species from Madagascar that inhabits the semiarid spiny forest of the southern part of the island. Habitat destruction and illegal harvesting greatly threaten this species, as attested by the significant reduction of its distribution area in the past 30 years. In order to undertake appropriate conservation actions, it is essential to acquire a better knowledge of its genetic structure. For this study, 145 blood samples were collected from three populations of radiated tortoises in southwestern Madagascar. Eight microsatellite loci were found to be polymorphic, and allelic diversity and observed heterozygosity were high for all markers.     

Population genomics of wild and laboratory zebrafish (Danio rerio)

Understanding a wider range of genotype–phenotype associations can be achieved through ecological and evolutionary studies of traditional laboratory models. Here, we conducted the first large‐scale geographic analysis of genetic variation within and among wild zebrafish (Danio rerio) populations occurring in Nepal, India, and Bangladesh, and we genetically compared wild populations to several commonly used lab strains. We examined genetic variation at 1832 polymorphic EST‐based single nucleotide polymorphisms (SNPs) and the cytb mitochondrial gene in 13 wild populations and three lab strains. Natural populations were subdivided into three major mitochondrial DNA clades with an average among‐clade sequence divergence of 5.8%. SNPs revealed five major evolutionarily and genetically distinct groups with an overall F_ST of 0.170 (95% CI 0.105–0.254). These genetic groups corresponded to discrete geographic regions and appear to reflect isolation in refugia during past climate cycles. We detected 71 significantly divergent outlier loci (3.4%) and nine loci (0.5%) with significantly low F_ST values. Valleys of reduced heterozygosity, consistent with selective sweeps, surrounded six of the 71 outliers (8.5%). The lab strains formed two additional groups that were genetically distinct from all wild populations. An additional subset of outlier loci was consistent with domestication selection within lab strains. Substantial genetic variation that exists in zebrafish as a whole is missing from lab strains that we analysed. A combination of laboratory and field studies that incorporates genetic variation from divergent wild populations along with the wealth of molecular information available for this model organism provides an opportunity to advance our understanding of genetic influences on phenotypic variation for a vertebrate species.