Microsatellites from the vairone Leuciscus souffia (Pisces: Cyprinidae) and their application to closely related species

Eleven microsatellites were isolated from the vairone Leuciscus souffia (Risso 1826), an endangered fish that inhabits river systems in and around the Alps in Europe. The level of genetic diversity was assessed in 29 individuals of the subspecies L. s. souffia, and their variability was further estimated in seven individuals of a different subspecies, L. s. muticellus. Eight of these microsatellite loci were also applied to seven closely related cyprinid species. Availability of the reported microsatellite loci will facilitate the investigation of population genetic structure of these species with applications for the development of conservation strategies and phylogeographical approaches.     

B Lymphocyte Colony Formation in vitro: Ultrastructural Development of Individual Colonies

B lymphocyte colony development in agar culture was studied using an electron microscope, and more than 3,000 colony cells were identified and photographed. In early cultures (day 4) lymphoblasts dominated the colonies. From day 5 onwards plasmablasts and small lymphocytes were present in the colonies. From day 6 onward mature plasma cells were observed in increasing numbers. On day 9 of culture the colonies started to degenerate and on day 10 of culture approximately 70% of the colony consisted of pyknotic and degenerating cells. Topographically, the degenerating cells were concentrated in the center of the colony whereas proliferation took place in the periphery. Colony growth occurred in an exponential fashion, the number of viable colony cells being maximal on day 8 of culture (400–600 cells/colony). At this time the frequencies of the four B cell categories were: lymphoblasts 72%, plasmablasts 20%, plasma cells 6%, and small lymphocytes 2%. Recloning experiments showed that dispersed colony cells were capable of forming only small cell clusters. It is concluded that B lymphocyte colony formation reflects a series of B cell developmental stages including the formation of the end cell categories of this lymphocyte lineage.     

Mercator: a fast and simple web server for genome scale functional annotation of plant sequence data

Next‐generation technologies generate an overwhelming amount of gene sequence data. Efficient annotation tools are required to make these data amenable to functional genomics analyses. The Mercator pipeline automatically assigns functional terms to protein or nucleotide sequences. It uses the MapMan ‘BIN’ ontology, which is tailored for functional annotation of plant ‘omics’ data. The classification procedure performs parallel sequence searches against reference databases, compiles the results and computes the most likely MapMan BINs for each query. In the current version, the pipeline relies on manually curated reference classifications originating from the three reference organisms (Arabidopsis, Chlamydomonas, rice), various other plant species that have a reviewed SwissProt annotation, and more than 2000 protein domain and family profiles at InterPro, CDD and KOG. Functional annotations predicted by Mercator achieve accuracies above 90% when benchmarked against manual annotation. In addition to mapping files for direct use in the visualization software MapMan, Mercator provides graphical overview charts, detailed annotation information in a convenient web browser interface and a MapMan‐to‐GO translation table to export results as GO terms. Mercator is available free of charge via http://mapman.gabipd.org/web/guest/app/Mercator .     

Ecological and evolutionary consequences of the trophic polymorphism in Cichlasoma citrinellum (Pisces: Cichlidae)

The neotropical cichlid fish Cichlasoma citrinellum is polymorphic in the structure of its pharyngeal jaw apparatus and external morphology. The pharyngeal jaws are either gracile and bear slender, pointed teeth (papilliform) or robust with strong, rounded teeth (molariform). Molariform morphs have a ‘benthic’, and papilliform morphs a ‘limnetic’ body form. Furthermore, this species is also polychromatic, with yellow and black morphs. The molariform morphology of the pharyngeal jaw apparatus adapts the fish for cracking and feeding on snails. Based on analysis of stomach contents, 94% of the molariform morph ate snails whereas only 19%, of the papilliform morph did so. This result suggests that the morphs occupy different ecological niches. The morphology of the pharyngeal jaw apparatus does not correlate significantly with sex, but it does with body colouration (P<0.005). Cichlasoma citrinellum mate assortatively with their own colour; therefore a mating preference for colour may lead to genetic isolation of trophic morphs. The frequency of the molariform morph differs strikingly among populations of five Nicaraguan lakes and its abundance is correlated with the abundance of snails, the fishes' principal prey item. Among populations the frequency of molariform morphs decreases in the dry season. Morphology possibly changes reversibly within particular individuals between seasons. These results suggest that phenotypic plasticity and polymorphisms may be an adaptive characteristic of cichlid fishes. Patterns of intraspecific morphological variation match patterns of interspecific morphological diversification which suggests that universal developmental mechanisms canalize the possible expressions of morphology. The ability to respond morphologically to environmental shifts, in conjunction with genetically determined trophic polymorphisms and sexual selection via mate choice, could be the basis for speciation through intermediate stages of polymorphism of the impressive adaptive radiation of cichlid fishes.     

Natural colonization or introduction? Phylogeographical relationships and morphological differentiation of house geckos (Hemidactylus) from Madagascar

Several house gecko species of the genus Hemidactylus are almost cosmopolitan lizards, with distributions that have probably been shaped by natural transoceanic dispersal as well as by more recent human introductions. Here we revise the Hemidactylus populations of Madagascar and compare them genetically with populations from other sites in the Indian Ocean region. Morphological data strongly confirm the occurrence of three Hemidactylus species on Madagascar: Hemidactylus frenatus , distributed along the western coast of Madagascar; H. platycephalus , restricted to the north-west and the widespread H. mercatorius that occurs throughout the island, including coastal areas at sea level as well as big cities (Antananarivo, Fianarantsoa) at altitudes of 1200–1300 m above sea level. Analyses of partial sequences of the 16S rRNA gene in 46 Hemidactylus specimens from Madagascar, East Africa, South Asia, and the Comoro and Mascarene archipelagos demonstrated the presence of a fourth species, H. brooki, on the Mascarenes (Réunion, Rodrigues, and Mauritius) and Comoros (Moheli). The Malagasy populations of H. platycephalus were genetically uniform and differentiated from the African and Comoroan specimens studied. H. frenatus had a relatively low genetic differentiation over the whole region with no recognizable phylogeographical structure, indicating more recent colonizations or introductions. In contrast, H. mercatorius showed a strong phylogeographical structure of haplotypes, with two distinctly different lineages in Madagascar. Moreover, all Malagasy specimens differed strongly from the single African specimen included. This indicates that populations of H. mercatorius in Madagascar have a long history that predates human settlement.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 83 , 115–130.     

Reduction of molecular gas diffusion through gaskets in leaf gas exchange cuvettes by leaf‐mediated pores

There is an ongoing debate on how to correct leaf gas exchange measurements for the unavoidable diffusion leakage that occurs when measurements are done in non‐ambient CO₂ concentrations. In this study, we present a theory on how the CO₂ diffusion gradient over the gasket is affected by leaf‐mediated pores (LMP) and how LMP reduce diffusive exchange across the gaskets. Recent discussions have so far neglected the processes in the quasi‐laminar boundary layer around the gasket. Counter intuitively, LMP reduce the leakage through gaskets, which can be explained by assuming that the boundary layer at the exterior of the cuvette is enriched with air from the inside of the cuvette. The effect can thus be reduced by reducing the boundary layer thickness. The theory clarifies conflicting results from earlier studies. We developed leaf adaptor frames that eliminate LMP during measurements on delicate plant material such as grass leaves with circular cross section, and the effectiveness is shown with respiration measurements on a harp of Deschampsia flexuosa leaves. We conclude that the best solution for measurements with portable photosynthesis systems is to avoid LMP rather than trying to correct for the effects.     

Multiple effects of temperature,photoperiod and food quality on the performance of a pine sawfly

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Microsatellites from the burbot (Lota lota), a freshwater gadoid fish (Teleostei)

We developed 21 polymorphic dinucleotide microsatellite loci, (CA)_n and (CT)_n, for the Holarctic freshwater fish, Lota lota, using an enriched genomic library protocol. The species has an interesting life history because winter‐spawning adults migrate over long distances to form spawning aggregations, a behaviour which should maintain genetic homogeneity across large spatial scales. Availability of the reported microsatellites will facilitate the investigation of population genetic structure with regard to postglacial colonization history and conservation strategies. The primers were screened on 30 individuals from a natural population (Lake Constance, southern Germany), revealing three to 24 alleles per locus with expected heterozygosities ranging from 0.48 to 0.93.