Separation and Purification of Protoplast Types from Commelina communis L. Leaf Epidermis

Guard cell and epidermal/subsidiary cell protoplasts obtainedby enzymic digestion of peeled Commelina communis leaf epidermiswere separated and purified by discontinuous density gradientccntrifugation with media based on Percoll (Pharmacia Fine ChemicalsAB, Uppsala, Sweden). The cell types were recovered over 99.9%pure at yields exceeding 50% efficiency, and mesophyll contaminationcould be virtually eliminated when desired. Osmotic characteristicsof the protoplast types were evaluated and compared to in vivovalues, and the viability of the protoplasts, assessed usinga range of criteria, was found to be high. Purified Commelinaguard cell protoplasts were able to evolve O₂ when illuminated,and this was substantially reduced in the presence of the inhibitorDCMU, indicating that they possess photosystem II activity.Specific advantages of this method of protoplast purification,and the potential uses of separate suspensions of guard cellsand epidermal/subsidiary cells in experiments on stomatal physiologyare discussed. Key words: Commelina communis, Protoplasts, Epidermis     

An Electron Microscope-Cytochemical Method for Differentiating Membranes of Host Red Cells and Malaria Parasites*

Limiting membranes of malaria parasites and host red cells stain differently when exposed to positively charged iron colloid. Negatively charged red cell membranes avidly bind colloid, whereas parasite membranes do not. This selectivity in colloidal iron uptake by the 2 types of membranes can be utilized as an aid in discerning the amounts of contaminating host cell membranes in “free” malaria parasite preparations and in related cell-free membrane extracts.     

The Structure and Development of the Reproductive Organs and Carposporophyte in two British Species of Gelidium

The structure and development of the reproductive organs andcarposporophyte are described for two British species of Gelidium,G. latifolium (Grev.) Born. & Thur. and G. pulchellum (Turn.)Kütz. The gonimoblast does not develop directly from theunchanged carpogonium, as was thought by Kylin, but a swollenmultinucleate cell of irregular outline is formed first, eitherfrom the carpogonium alone, or by the fusion of the carpogoniumand certain neighbouring cells; the gonimoblast develops fromlong, non-septate processes of this multi-nucleate cell. Thesignificance of the fusion of the carpogonium and certain vegetativecells is discussed in relation to the various definitions ofthe auxiliary cell which have been proposed. It is concludedthat an auxiliary cell does not occur in Gelidium.     

Hazards Affecting Grizzly Bear Survival in the Greater Yellowstone Ecosystem

Abstract: During the past 2 decades, the grizzly bear (Ursus arctos) population in the Greater Yellowstone Ecosystem (GYE) has increased in numbers and expanded its range. Early efforts to model grizzly bear mortality were principally focused within the United States Fish and Wildlife Service Grizzly Bear Recovery Zone, which currently represents only about 61% of known bear distribution in the GYE. A more recent analysis that explored one spatial covariate that encompassed the entire GYE suggested that grizzly bear survival was highest in Yellowstone National Park, followed by areas in the grizzly bear Recovery Zone outside the park, and lowest outside the Recovery Zone. Although management differences within these areas partially explained differences in grizzly bear survival, these simple spatial covariates did not capture site-specific reasons why bears die at higher rates outside the Recovery Zone. Here, we model annual survival of grizzly bears in the GYE to 1) identify landscape features (i.e., foods, land management policies, or human disturbances factors) that best describe spatial heterogeneity among bear mortalities, 2) spatially depict the differences in grizzly bear survival across the GYE, and 3) demonstrate how our spatially explicit model of survival can be linked with demographic parameters to identify source and sink habitats. We used recent data from radiomarked bears to estimate survival (1983–2003) using the known-fate data type in Program MARK. Our top models suggested that survival of independent (age ≥ 2 yr) grizzly bears was best explained by the level of human development of the landscape within the home ranges of bears. Survival improved as secure habitat and elevation increased but declined as road density, number of homes, and site developments increased. Bears living in areas open to fall ungulate hunting suffered higher rates of mortality than bears living in areas closed to hunting. Our top model strongly supported previous research that identified roads and developed sites as hazards to grizzly bear survival. We also demonstrated that rural homes and ungulate hunting negatively affected survival, both new findings. We illustrate how our survival model, when linked with estimates of reproduction and survival of dependent young, can be used to identify demographically the source and sink habitats in the GYE. Finally, we discuss how this demographic model constitutes one component of a habitat-based framework for grizzly bear conservation. Such a framework can spatially depict the areas of risk in otherwise good habitat, providing a focus for resource management in the GYE.     

Genetic diversity and differentiation at MHC genes in island populations of tuatara (Sphenodon spp.)

Neutral genetic markers are commonly used to understand the effects of fragmentation and population bottlenecks on genetic variation in threatened species. Although neutral markers are useful for inferring population history, the analysis of functional genes is required to determine the significance of any observed geographical differences in variation. The genes of the major histocompatibility complex (MHC) are well‐known examples of genes of adaptive significance and are particularly relevant to conservation because of their role in pathogen resistance. In this study, we survey diversity at MHC class I loci across a range of tuatara populations. We compare the levels of MHC variation with that observed at neutral microsatellite markers to determine the relative roles of balancing selection, diversifying selection and genetic drift in shaping patterns of MHC variation in isolated populations. In general, levels of MHC variation within tuatara populations are concordant with microsatellite variation. Tuatara populations are highly differentiated at MHC genes, particularly between the northern and Cook Strait regions, and a trend towards diversifying selection across populations was observed. However, overall our results indicate that population bottlenecks and isolation have a larger influence on patterns of MHC variation in tuatara populations than selection.     

Systematic revision of Hoggicosa Roewer, 1960, the Australian ‘bicolor’ group of wolf spiders (Araneae: Lycosidae)

The Australian wolf spider genus Hoggicosa Roewer, 1960 with the type species Hoggicosa errans (Hogg, 1905) is revised to include ten species: Hoggicosa alfi sp. nov. ; Hoggicosa castanea (Hogg, 1905) comb. nov. (= Lycosa errans Hogg, 1905 syn. nov. ; = Lycosa perinflata Pulleine, 1922 syn. nov. ; = Lycosa skeeti Pulleine, 1922 syn. nov. ); Hoggicosa bicolor (McKay, 1973) comb. nov. ; Hoggicosa brennani sp. nov. ; Hoggicosa duracki (McKay, 1975) comb. nov. ; Hoggicosa forresti (McKay, 1973) comb. nov. ; Hoggicosa natashae sp. nov. ; Hoggicosa snelli (McKay, 1975) comb. nov. ; Hoggicosa storri (McKay, 1973) comb. nov. ; and Hoggicosa wolodymyri sp. nov. The Namibian Hoggicosa exigua Roewer, 1960 is transferred to Hogna, Hogna exigua (Roewer, 1960) comb. nov. A phylogenetic analysis including nine Hoggicosa species, 11 lycosine species from Australia and four from overseas, with Arctosa cinerea Fabricius, 1777 as outgroup, supported the monophyly of Hoggicosa, with a larger distance between the epigynum anterior pockets compared to the width of the posterior transverse part. The analysis found that an unusual sexual dimorphism for wolf spiders (females more colourful than males), evident in four species of Hoggicosa, has evolved multiple times. Hoggicosa are burrowing lycosids, several constructing doors from sand or debris, and are predominantly found in semi‐arid to arid regions of Australia. © 2010 The Linnean Society of London, Zoological Journal of the Linnean Society, 2010, 158 , 83–123.     

Influence of ultraviolet-B radiation on the drift response of stream invertebrates

1. Recent studies have shown that ultraviolet (UV) radiation (280–400 nm) has increased by ≈ 8% in temperate regions over the past decade, but little effort has been devoted to understanding the ecological effects on temperate ecosystems. This research examined the effects of artificial ultraviolet-B (UVB; 280–320 nm) radiation on the drift response of immature stream insects in laboratory microcosms.
2. Two experiments involved natural populations of stream invertebrates, collected from the Cache la Poudre River (September 1994) and the Arkansas River (October 1995) in Colorado. UVB lamps were turned on from 10.00 to 14.00 h each day, and drifting animals were collected on days 1, 3, 5 and 7 during the exposure period. Levels of artificial UVB used in these experiments were similar to levels that stream organisms experience during clear, mid-day conditions at Fort Collins, Colorado (longitude 105°30'; latitude 40°35').
3. Drift was significantly higher in microcosms exposed to UVB than in controls and was dominated by Baetis sp. (Ephemeroptera: Baetidae), Trichoptera (caddisflies) and Simulium sp. (Diptera: Simuliidae). The increased drift of some stream invertebrates in UVB-exposed streams may be a behavioural response and/or a result of injury.
4. Stream organisms may be particularly sensitive to predicted increases in UV radiation, because streams are generally shallow with clear water. As a result of this potential sensitivity, we recommend that research be directed to understanding the ecological effects of UV radiation on these habitats.     

Soil-surface carbon dioxide efflux and microbial biomass in relation to tree density 13 years after a stand replacing fire in a lodgepole pine ecosystem

The effects of fire on soil‐surface carbon dioxide (CO₂) efflux, F_S, and microbial biomass carbon, C_mic, were studied in a wildland setting by examining 13‐year‐old postfire stands of lodgepole pine differing in tree density (< 500 to > 500 000 trees ha^?1) in Yellowstone National Park (YNP). In addition, young stands were compared to mature lodgepole pine stands (～110‐year‐old) in order to estimate ecosystem recovery 13 years after a stand replacing fire. Growing season F_S increased with tree density in young stands (1.0 µmol CO₂ m^?2 s^?1 in low‐density stands, 1.8 µmol CO₂ m^?2 s^?1 in moderate‐density stands and 2.1 µmol CO₂ m^?2 s^?1 in high‐density stands) and with stand age (2.7 µmol CO₂ m^?2 s^?1 in mature stands). Microbial biomass carbon in young stands did not differ with tree density and ranged from 0.2 to 0.5 mg C g^?1 dry soil over the growing season; C_mic was significantly greater in mature stands (0.5–0.8 mg C g^?1 dry soil). Soil‐surface CO₂ efflux in young stands was correlated with biotic variables (above‐ground, below‐ground and microbial biomass), but not with abiotic variables (litter and mineral soil C and N content, bulk density and soil texture). Microbial biomass carbon was correlated with below‐ground plant biomass and not with soil carbon and nitrogen, indicating that plant activity controls not only root respiration, but C_mic pools and overall F_S rates as well. These findings support recent studies that have demonstrated the prevailing importance of plants in controlling rates of F_S and suggest that decomposition of older, recalcitrant soil C pools in this ecosystem is relatively unimportant 13 years after a stand replacing fire. Our results also indicate that realistic predictions and modeling of terrestrial C cycling must account for the variability in tree density and stand age that exists across the landscape as a result of natural disturbances.