Systematics and phylogeny of the tribe Paragini (Diptera: Syrphidae) based on molecular and morphological characters

We studied the phylogeny and systematics of the tribe Paragini (Diptera: Syrphidae) using morphological and molecular data. The paper presents separate parsimony analyses of both adult morphological characters and partial DNA sequence data from mitochondrial cytochrome c oxidase I and nuclear ribosomal 28S rRNA gene, as well as a combined analysis of all the data. The data set of morphological characters included some features of the male terminalia (i.e. shape of the ejaculatory apodeme; relative position of elements of the aedeagal complex; shape of surstylar apodeme; shape of the aedeagal apodeme) not previously used in the systematics of the Paragini. The trees obtained from separate parsimony analyses of molecular and morphological data produced almost identical topologies. Four lineages are supported by the combined data set, and we establish two new subgenera, i.e. Serratoparagus Vujić et Radenković subgen. nov., and Afroparagus Vujić et Radenković subgen. nov., and redefine Pandasyopthalmus Stuckenberg, 1954 stat. rev. and Paragus Latreille, 1804, stat. rev. The monophyly of the Pandasyopthalmus clade, including the species fitting neither of the current species groups ( jozanus -group) of Paragini, is established. Diagnoses of all known species groups are presented, including a new arrangement of almost all valid species of Paragini.  © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society , 2008, 152 , 507–536.     

Carbon dioxide exchange above a Mediterranean C3/C4 grassland during two climatologically contrasting years

Eddy‐covariance measurements of net ecosystem carbon exchange (NEE) were carried out above a grazed Mediterranean C3/C4 grassland in southern Portugal, during two hydrological years, 2004–2005 and 2005–2006, of contrasting rainfall. Here, we examine the seasonal and interannual variation in NEE and its major components, gross primary production (GPP) and ecosystem respiration (R_eco), in terms of the relevant biophysical controls. The first hydrological year was dry, with total precipitation 45% below the long‐term mean (669 mm) and the second was normal, with total precipitation only 12% above the long‐term mean. The drought conditions during the winter and early spring of the dry year limited grass production and the leaf area index (LAI) was very low. Hence, during the peak of the growth period, the maximum daily rate of NEE and the light‐use and water‐use efficiencies were approximately half of those observed in the normal year. In the summer of 2006, the warm‐season C4 grass, Cynodon dactylon L., exerted an evident positive effect on NEE by converting the ecosystem into a carbon sink after strong rain events and extending the carbon sequestration for several days, after the end of senescence of the C3 grasses. On an annual basis, the GPP and NEE were 524 and 49 g C m^?2, respectively, for the dry year, and 1261 and ?190 g C m^?2 for the normal year. Therefore, the grassland was a moderate net source of carbon to the atmosphere, in the dry year, and a considerable net carbon sink, in the normal year. In these 2 years of experiment the total amount of precipitation was the main factor determining the interannual variation in NEE. In terms of relevant controls, GPP and NEE were strongly related to incident photosynthetic photon flux density on short‐term time scales. Changes in LAI explained 84% and 77% of the variation found in GPP and NEE, respectively. Variations in R_eco were mainly controlled by canopy photosynthesis. After each grazing event, the reduction in LAI affected negatively the NEE.     

Distribution of Epitopes of Trypanosoma cruzi Amastigotes During the Intracellular Life Cycle within Mammalian Cells

ABSTRACT. In this study we have examined the distribution of epitopes defined by monoclonal antibodies raised against Trypanosoma cruzi amastigotes during the intraceullar life cycle of the parasite. We have raised monoclonal antibodies towards amastigote forms and performed preliminary immunochemical characterization of their reactivities. MAB 1D9, 3G8, 2B7, 3B9, and 4B9, and 4B9 react with carbohydrate epitopes of the parasite major surface glycoprotein—Ssp-4 defined by MAB 2C2 [5]: MAB 4B5 reacts with a noncarbohydrate epitope in all developmental stages of the parasite, and MAB 3B2 also detects a noncarbohydrate epitope preferentially in T. cruzi flagellared forms. Vero cells infected with tissue culture-derived trypomastigotes of clone D11 (G strain) were fixed at different times during the intraceullular proliferation of parasites, and processed for immjno-electron microscopy and confocal immunoflurescence with the different monoclonal antibodies. We observed that while the surface distribution of MAB 2C2 and 4B9 epitopes was uniform throughout the cycle, MAB 1D9, 3G8, and 2B7 reacted with cytoplasmic membrance-bound compartments of the amastigotes. MAB 3B9 displayed a unique surface dentate pattern in some amastigotes. MAB 4B5 recognized a curved-shaped structure at the flagellar pocket region in some intracellular amastigotes and localized to the membrane in dividing forms. In intracellular trypomastigotes, MAB 4B5 also displayed a punctate pattern near the flagellar pocket.     

Release of Membrane-Bound Trails by Trypanosoma cruzi Amastigotes onto Modified Surfaces and Mammalian Cells

ABSTRACT. Upon incubation at 37° C onto glass coverslips coated with Concanavalin A, poly-L-lysine, or a monoclonal antibody (1D9) directed to the parasite major surface glycoprotein Ssp-4, extracellular Trypanosoma cruzi amastigotes release trails of material barely visible by light microscopy. This release is not associated with parasite movements. Immunolabeling studies confirmed that the material is derived from the parasite's membrane since thin section through samples labeled with 1D9 revealed that the trails are membrane-bound structures. Scanning electron microscopy showed that the ∼0.1-μm thick trails of material emerging from the amastigotes can be uniform or beaded, indicating a tendency to vesiculation. The trails are preferentially released from the flagellar pocket region and/or at the opposite posterior end of the parasite body, and seem to be devoid of microtubules. The release is time and temperature-dependent and fixed parasites do not form trails. All attempts to inhibit trail release using drugs (antimycin A, sodium azide, cytochalasin D, nocodazole, genistein, staurosporine, EGTA) failed. The observation of trails associated with intracellular parasites and amastigotes invading Vero cells suggests that this is probably a physiological process.     

Nitrogen nutrition and the level of Crassulacean acid metabolism in Kalanchoë lateritia Engl.

Abstract. The influence of different nitrogen levels was studied in the CAM facultative Kalanchoë lateritia by watering some plants with Hoagland's solution (which contains, besides other ions, NO₃^?= 14.47mol m^?3and NH₄⁺= 1.04mol m^?3; N group), and others with the same solution but with combined nitrogen concentration reduced to either one fifth (NO₃^?= 2.894mol m^?3 and NH₄⁺= 0.208mol m^?3; N/5 group) or one tenth (NO₃^?= 1.447mol m^?3and NH₄⁺= 0.104mol m-³;N/10 group). The influence of the three nitrogen levels on CAM expression was assessed through activities of PEP-Case, PPD and RuBisCo, diurnal fluctuation of titratable acidity, mesophyll succulence, soluble protein, chlorophyll, nitrate, starch contents, pattern of nocturnal CO₂ exchange and electron microscopy. CAM photosynthesis was more intense in N/5 plants which also had the highest Sm value. The activity of RuBisCo showed no significant differences in the three situations (expressed on chlorophyll basis) whereas both PEP-Case and PPD had higher values in N/5” plants. Chlorophyll and soluble protein were more abundant in the N plants followed by N/5 and N/10 plants. Nitrate was higher in N plants and starch content in N/5 plants. IRGA determination of CO₂ nocturnal uptake showed that N/5 plants began CO₂ capture earlier and at a more intense rate and for a longer period than plants from other groups also having a daily variation of titratable acidity (97.71 ± 10.8 μeq. G^?1 f.w.) indicative of performing strong CAM. Electron microscope morphometric analysis revealed larger chloroplasts in N plants and smaller in N/10 plants, with starch fractional volume higher in N/5 plants, correlating with more intense CAM activity of these plants.     

A review of the genus Semele (Ruscaceae) systematics in Madeira

The present study attempts to review the systematics of Semele (Ruscaceae) in Madeira, based on phenotypic diversity. The variation in some vegetative (climbing shoot, second-order branches or 'phylloclades') and sexual (inflorescence and flowers) characters was analysed in 115 plant specimens from 30 field populations, herbaria of the Costa collection and Madeira Botanical Garden (MADJ) and certain gardens. Thirty-one quantitative and qualitative characters have been utilized in the analysis. Kaiser–Meyer–Olkin (KMO) analysis indicates adequate sampling. Principal component analysis (PCA) reveals that the spatial distribution of individuals has a discontinuous behaviour. Principal coordinate analysis (PCO) utilizing the Gower coefficient on standardized data revealed a significantly discontinuous distribution of individuals, such that two different clusters can be defined. The Student's t -test and Tukey test on separate characters, when individuals were classified according to the Costa classification, confirms the significant differences between grouping accessions. This leads to the recognition of two species within the genus in Madeira. Literature and herbarium studies show that these two taxa are conspecific with Semele androgyna (L.) Kunth sensu stricto (s.s.) and Semele menezesi Costa sensu lato (s.l.) . A separated statistical analysis of the S. androgyna cluster shows the possible existence of additional subgroups. Based on field population distribution, ecological behaviour and variation in features, we propose the recognition of two species, S. androgyna (L.) Kunth and S. menezesi (Costa) Pinheiro de Carvalho, and two subspecies S. androgyna (L.) Kunth androgyna Pinheiro de Carvalho and S. androgyna (L.) Kunth pterygophora Pinheiro de Carvalho.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 483–497.     

Aquatic macrophytes diversity in lagoons of a tropical floodplain: The role of connectivity and water level

Abstract The goal of this research is to evaluate how the assemblages of aquatic macrophytes in marginal floodplain habitats, with different degrees of connectivity to the main river, respond to water level fluctuations. Samples were carried out quarterly (May 2000 to March 2002) in seven lagoons of the Upper Paraná River floodplain (22°30′ and 22°45′‐S and 53°15′ and 53°30′‐W) with different degrees of connectivity (connected and disconnected to the main river). In each lagoon, a shore‐pelagic zone transect was marked and at every 2 m the depth and the cover of each aquatic macrophyte species were recorded (Domin‐Krajina scale) in a quadrat of 0.25 m². A total of 29 aquatic macrophyte species and an unusual decrease in water level were recorded in August 2001. Drawdown had a negative impact on species richness, only in connected lagoons, which was shown by a positive relationship between depth and species number (r‐Spearman = 0.86; P < 0.01). Depth affected Beta diversity positively (r‐Spearman = 0.79; P < 0.05). Drawdown affected the connected and disconnected lagoons differently, which can be attributed to their different morphometry. In this period, ‘habitat contraction’ was higher on connected lagoons. Positive correlation between mean species number and depth, and between beta diversity and depth, are factors that support this affirmation. Indicator species analysis showed that for disconnected lagoons, Oxycaryum cubense (Cyperaceae), Polygonum meissnerianum (Polygonaceae) and P. ferrugineum, with indicator values (IndVal) of 53, 30 and 25%, respectively, were indicator species. Salvinia spp. (Salvinaceae) (62%), P. acuminatum (44%) and the Ricciaceae Ricciocarpus natans (0%) were the indicator species of the connected lagoons.     

Isolation of polymorphic tetranucleotide microsatellite markers for the silky short‐tailed bat Carollia brevicauda

We isolated 10 polymorphic microsatellite markers for the silky short‐tailed bat, Carollia brevicauda, from genomic libraries enriched for (AAGG)_n repetitive elements. The number of alleles ranged from six to 25 per locus with the observed heterozygosity ranging from 0.29 to 0.95. These markers will be useful for analysis of questions concerning population genetic structure and models of speciation. Results of cross‐species amplification in Carollia castanea and Carollia perspicillata are also reported.     

Isolation and characterization of polymorphic microsatellite loci in the scarlet ibis (Eudocimus ruber— Threskiornithidae‐Aves)

Here we presented 10 polymorphic microsatellite loci obtained from scarlet ibis through an enriched genomic library. The analysis of 45 individuals from three Brazilian natural populations showed allelic diversity ranged from three to 17 alleles, observed heterozygosity ranged from 0.03 to 0.92, and expected heterozygosity ranged from 0.06 to 0.92. These highly variable microsatellite loci can provide means for assessing overall genetic variation in its remnant natural populations, which may help the development of effective conservation programs.     

Further Characterization of Carbohydrate-Containing Fractions from Trypanosoma mega1

Epimastigotes of Trypanosoma mega were submitted to phenol extraction after lipid extraction, providing an extract whose carbohydrate portion (30%) contained fucose, ribose, xylose, mannose, galactose, and glucose. The purified fraction recovered in the void volume of Bio Gel P-150 gave on SDS-PAGE a band of M_r～ 55,000 positive for protein and carbohydrate and a diffuse band strongly positive for carbohydrate and lipids (M_r～ 22,000). The structural analysis of the carbohydrate moiety of this fraction by GLC-MS indicated the presence of nonreducing end groups of fucopyranose, mannopyranose, and galactopyranose, 3-O- and 4-O-substituted and 2,3- and 2,4-di-O-substituted galactopyranosyl units. Extraction of this fraction with chloroform/methanol/water provided a soluble fraction that on SDS-PAGE gave rise to a carbohydrate and lipid-positive band (M_r～ 22,000). This fraction contained fucose, mannose, and galactose (1:1:1). As main branch points, 2,3-di-O-substituted galactopyranosyl units were present according to methylation data. Similar proportions of fucopyranosyl, mannopyranosyl, galactopyranosyl end units were present. The presence of lipids in this fraction was confirmed by methanolysis following isolation and characterization of the corresponding fatty acid methyl esters. Palmitic acid (16:0) and an 18:1 fatty acid were the predominant fatty acids.