Molecular Markers for Ion Compartmentation in Cells of Higher Plants

The tonoplast plays a crucial role in ion compartmentation,which is a central feature of the salt tolerance of halophytes,but we do not know the properties of the membrane that conferthis ability. A method was, therefore, developed for the isolationof vacuoles from Suaeda maritima (L.) Dum. of sufficiently highpurity to enable biochemical characterization of their lipidand protein composition. Tonoplast fractions produced by densitygradient centrifugation, as well as vacuoles isolated by a varietyof methods (including DEAE dextran lysis, digitonin lysis, andmechanical shear forces) were unacceptably contaminated. A highlypure vacuole preparation was obtained when protoplasts werelysed by a mild hypotonic shock in alkaline buffer, in the presenceof the compatible cytosolute glycine-betaine, followed by shearforce during ultracentrifugation; cytoplasmic contaminationwas prevented by the addition of the zwitterionic detergent3-([3-cholamidopropyl]dimethylammonio)-l-propanesulphonate (CHAPS).Light microscopy of this preparation revealed no intact protoplastsand no contamination by chlorophyll could be detected. Electronmicroscopy showed the vacuoles to be single-membrane-bound structures,and was the only criterion upon which vacuoles could be separatedreliably from vacuoplasts, in which the plasmalemma is collapsedon to the tonoplast. Analysis by SDS-PAGE showed that a totalof 15 polypeptides were enriched in the tonoplast and 27 inthe soluble fraction from vacuole preparations, with a patternsimilar to that reported for glycophytic species. The pure tonoplastexhibited both vanadate-insensitive ATPase and pyrophosphataseactivities, but the properties of these enzymes were broadlysimilar to those of glycophytes. Analysis of membrane fattyacids showed that the degree of saturation of the putative tonoplastpreparation increased as the assessment of the purity of thepreparation (made by microscopy) increased. The ATPase couldbe substantially purified by ion-exchange FPLC. The resultsare discussed in relation to the degree of purity needed inmembrane preparations in order to be suitable for biochemicalanalysis. Key words: ATPase, membrane lipids, tonoplast, salinity, Suaeda maritima     

Univariate analysis of tsetse habitat in the common fly belt of Southern Africa using climate and remotely sensed vegetation data

Abstract Tsetse are vectors of trypanosomes that cause diseases both in humans and livestock. Traditional tsetse surveys, using sampling methods such as Epsilon traps and black screen fly rounds, are often logistically difficult, costly and time-consuming. The distribution of tsetse, as revealed by such survey methods, is strongly influenced by environmental conditions, such as climate and vegetation cover, which may be readily mapped using satellite data. These data may be used to make predictions of the probable distribution of tsetse in unsurveyed areas by determining the environmental characteristics of areas of tsetse presence and absence in surveyed areas. The same methods may also be used to characterize differences between tsetse species and subspecies. In this paper we analyse the distribution of Glossina morsitans centralis, Glossina morsitans morsitans and Glossina pallidipes in southern Africa with respect to single environmental variables. For G.m.centralis the best predictions were made using the average NDVI (75% correct predictions; range > 0.37) and the average of the maximum temperature (70% correct predictions; 27.0–29.2°C). For G.m.morsitans the best prediction was given by the maximum of the minimum temperature (84% correct predictions; range > 18.8°C), and for G.pallidipes , also by the maximum of the minimum temperature (86% correct predictions; range > 19.6 °C). The following paper compares a range of multivariate techniques for making predictions about the distribution of these species in the same region.     

Elevated atmospheric CO2 differentially affects needle chloroplast ultrastructure and phloem anatomy in Pinus palustris: interactions with soil resource availability

The response of forest species to increasing atmospheric CO₂, particularly under resource limitations, will require study in order to predict probable changes which may occur at the plant, community and ecosystem levels. Longleaf pine (Pinus palustris Mill.) seedlings were grown for 20 months at two levels of CO₂ (365 and 720 μol mol¹) in two levels of soil nitrogen (4 and 40 g m^?2), and with two levels of soil moisture (–0·5 and –1·5 MPa xylem pressure potential). Leaf tissue was collected in the spring (12 months exposure) and autumn (20 months exposure) and examined using transmission electron microscopy (TEM) and light microscopy. During early spring, elevated CO₂ magnified effects of N and water treatment on starch accumulation and in some cases contributed to altered organization of mesophyll chloroplasts. Disruption of chloroplast integrity was pronounced under elevated CO₂, low N and water stress. In autumn, needles contained little starch; however, chloroplasts grown under high CO₂ exhibited stress symptoms including increased plastoglobuli and shorter grana. A trend for reduced needle phloem cross-sectional area resulting from fewer sieve cells was also observed under elevated CO₂. These results suggest that, in nature, longleaf pine seedlings may not benefit from a doubling of CO₂, especially when soil resources are limiting.     

Linking riparian vegetation types and fluvial geomorphology along the Sabie River within the Kruger National Park, South Africa

Vegetation types were studied in relation to the fluvial geomorphology along the mixed bedrock‐alluvial Sabie River within the Kruger National Park, Mpumalanga, South Africa. Six vegetation types were identified using TWINSPAN analysis, namely: Phragmites mauritianus , Phyllanthus reticulatus , Breonadia salicina , Combretum erythrophyllum , Diospyros mespiliformis and Spirostachys africana vegetation types. Spirostachys africana and Diospyros mespiliformis vegetation types were found to occur predominantly on the stable, infrequently flooded macro‐channel banks, while the remaining four vegetation types were found almost exclusively along the more geomorphically and hydrologically dynamic macro‐channel floor. The degree of bedrock or alluvial influence was identified as being an integral factor in the distribution of the four macro‐channel floor vegetation types at both the morphological unit and the channel type scale. The geomorphological continuum from the bedrock influenced bedrock anastomosing channel types, to mixed anastomosing and pool‐rapid channel types, to the fully alluvial braided channel types, is reflected in the change in species composition from Breonadia salicina vegetation type, to Phyllanthus reticulatus and Phragmites mauritianus vegetation types, to Combretum erythrophyllum vegetation types, respectively. Given the vegetation/fluvial geomorphology links established, changes in vegetation composition are proposed in response to scenarios of geomorphological change as a result of progressive sedimentation.     

Effects of elevated atmospheric CO2 in agro-ecosystems on soil carbon storage

Increasing global atmospheric CO₂ concentration has led to concerns regarding its potential effects on the terrestrial environment. Attempts to balance the atmospheric carbon (C) budget have met with a large shortfall in C accounting (≈1.4 × 10¹⁵ g C y^–1) and this has led to the hypothesis that C is being stored in the soil of terrestrial ecosystems. This study examined the effects of CO₂ enrichment on soil C storage in C3 soybean (Glycine max L.) Merr. and C4 grain sorghum (Sorghum bicolor L.) Moench. agro-ecosystems established on a Blanton loamy sand (loamy siliceous, thermic, Grossarenic Paleudults). The study was a split-plot design replicated three times with two crop species (soybean and grain sorghum) as the main plots and two CO₂ concentration (ambient and twice ambient) as subplots using open top field chambers. Carbon isotopic techniques using δ¹³C were used to track the input of new C into the soil system. At the end of two years, shifts in δ¹³C content of soil organic matter carbon were observed to a depth of 30 cm. Calculated new C in soil organic matter with grain sorghum was greater for elevated CO₂ vs. ambient CO₂ (162 and 29 g m^–2, respectively), but with soybean the new C in soil organic matter was less for elevated CO₂ vs. ambient CO₂ (120 and 291 g m^–2, respectively). A significant increase in mineral associated organic C was observed in 1993 which may result in increased soil C storage over the long-term, however, little change in total soil organic C was observed under either plant species. These data indicate that elevated atmospheric CO₂ resulted in changes in soil C dynamics in agro-ecosystems that are crop species dependent.     

Effects of nutrient loading and extreme rainfall events on coastal tallgrass prairies: invasion intensity, vegetation responses, and carbon and nitrogen distribution

Soil fertility and precipitation are major factors regulating transitions from grasslands to forests. Biotic regulation may influence the effects of these abiotic drivers. In this study, we examined the effects of extreme rainfall events, anthropogenic nutrient loading and insect herbivory on the ability of Chinese tallow tree ( Sapium sebiferum ) to invade coastal prairie to determine how these factors may influence woody invasion of a grassland. We manipulated soil fertility (NPK addition) and simulated variation in frequency of extreme rainfall events in a three growing season, full factorial field experiment. Adding water to or pumping water out of plots simulated increased and decreased rainfall frequencies. We added Sapium seeds and seedlings to each plot and manipulated insect herbivory on transplanted Sapium seedlings with insecticide. We measured soil moisture, Sapium performance, vegetation mass, and carbon and nitrogen in vegetation and soils (0–10 cm deep, 10–20 cm deep). Fertilization increased Sapium invasion intensity by increasing seedling survival, height growth and biomass. Insect damage was low and insect suppression had little effect in all conditions. Recruitment of Sapium from seed was very low and independent of treatments. Vegetation mass was increased by fertilization in both rainfall treatments but not in the ambient moisture treatment. The amount of carbon and nitrogen in plants was increased by fertilization, especially in modified moisture plots. Soil carbon and nitrogen were independent of all treatments. These results suggest that coastal tallgrass prairies are more likely to be impacted by nutrient loading, in terms of invasion severity and nutrient cycling, than by changes in the frequency of extreme rainfall events.     

Burrow and mate fidelity in the Little Penguin Eudyptula minor at Lion Island, New South Wales, Australia

Although both mate and nest fidelity are thought to exist in Little Penguins Eudyptula minor , few studies have statistically assessed the relationship between both types of fidelity and breeding success. Observations of the breeding biology of the Little Penguin were carried out between 1990 and 1998 at Lion Island, New South Wales, Australia. The existence of fidelity and the effect of breeding success on subsequent fidelity were assessed for this population. Strong evidence for the existence of nest and mate fidelity was found with high rates of return to the same burrow and mate being recorded for both sexes. For all years of the study, the average number of female and male birds faithful to their nest was 76 and 79%, respectively, with only two noted cases of intraseasonal burrow swapping. The rate of divorce varied widely between years, with a range of 0–40%. A marginal association of a log linear model showed a highly significant relationship ( P  < 0.001) between success of a clutch in one year and fidelity to a mate in the following year. Consequently, parents successfully raising a fledgling in one year were more likely to reunite in the following year than were pairs that attempted unsuccessfully to breed. Pairs failing to fledge chicks were much more likely to separate owing to the absence of one parent in the following year. We were unable to detect any association between breeding success in one year and subsequent change of nest in the next.     

Freezing of Plant Tissues3

The freezing of plant cells under the microscope was studiedon three types of tissues: Tradescantia staminal hairs, fruitskin, and moss leaf. Cinemicrography was used to record stagesin the freezing processes, which are otherwise un observable.The following phenomena have been demonstrated and discussed:(1) The protective effect of persistent supercooling. (2) Ice-inoculationof cells and factors affecting it. (3) Sequence of ice formationwithin cells. (4) Modes of ice formation as affected by supercooling.(5) Freezing of cell walls and their longitudinal splitting.     

Proteomic and phenotypic profiling of the amphibian pathogen Batrachochytrium dendrobatidis shows that genotype is linked to virulence

Population genetics of the amphibian pathogen Batrachochytrium dendrobatidis ( Bd ) show that isolates are highly related and globally homogenous, data that are consistent with the recent epidemic spread of a previously endemic organism. Highly related isolates are predicted to be functionally similar due to low levels of heritable genetic diversity. To test this hypothesis, we took a global panel of Bd isolates and measured (i) the genetic relatedness among isolates, (ii) proteomic profiles of isolates, (iii) the susceptibility of isolates to the antifungal drug caspofungin, (iv) the variation among isolates in growth and phenotypic characteristics, and (v) the virulence of isolates against the European common toad Bufo bufo . Our results show (i) genotypic differentiation among isolates, (ii) proteomic differentiation among isolates, (iii) no significant differences in susceptibility to caspofungin, (iv) differentiation in growth and phenotypic/morphological characters, and (v) differential virulence in B. bufo . Specifically, our data show that Bd isolates can be profiled by their genotypic and proteomic characteristics, as well as by the size of their sporangia. Bd genotypic and phenotypic distance matrices are significantly correlated, showing that less-related isolates are more biologically unique. Mass spectrometry has identified a set of candidate genes associated with inter-isolate variation. Our data show that, despite its rapid global emergence, Bd isolates are not identical and differ in several important characters that are linked to virulence. We argue that future studies need to clarify the mechanism(s) and rate at which Bd is evolving, and the impact that such variation has on the host–pathogen dynamic.