Photosynthetic limitations and volatile and non‐volatile isoprenoids in the poikilochlorophyllous resurrection plant Xerophyta humilis during dehydration and rehydration

We investigated the photosynthetic limitations occurring during dehydration and rehydration of Xerophyta humilis, a poikilochlorophyllous resurrection plant, and whether volatile and non‐volatile isoprenoids might be involved in desiccation tolerance. Photosynthesis declined rapidly after dehydration below 85% relative water content (RWC). Raising intercellular CO₂ concentrations during desiccation suggest that the main photosynthetic limitation was photochemical, affecting energy‐dependent RuBP regeneration. Imaging fluorescence confirmed that both the number of photosystem II (PSII) functional reaction centres and their efficiency were impaired under progressive dehydration, and revealed the occurrence of heterogeneous photosynthesis during desiccation, being the basal leaf area more resistant to the stress. Full recovery in photosynthetic parameters occurred on rehydration, confirming that photosynthetic limitations were fully reversible and that no permanent damage occurred. During desiccation, zeaxanthin and lutein increased only when photosynthesis had ceased, implying that these isoprenoids do not directly scavenge reactive oxygen species, but rather protect photosynthetic membranes from damage and consequent denaturation. X. humilis was found to emit isoprene, a volatile isoprenoid that acts as a membrane strengthener in plants. Isoprene emission was stimulated by drought and peaked at 80% RWC. We surmise that isoprene and non‐volatile isoprenoids cooperate in reducing membrane damage in X. humilis, isoprene being effective when desiccation is moderate while non‐volatile isoprenoids operate when water deficit is more extreme.     

Inheritance and expression of introduced DNA in transgenic onion plants (Allium cepa)

Transgenic onion plants (Allium cepa) containing the Cauliflower mosaic virus 35s promoter (CaMV35s) and gfp gene construct encoding the visual green fluorescent reporter protein from pBINm gfp ER and the CaMV35s‐bar gene construct encoding resistance to the herbicide phosphinothricin from pCAMBlA3301 were produced by Agrobacterium‐mediated transformation. These plants weregrown to maturity and selfed in order to determine the expression and inheritance of the transgenes. CaMV35s regulation in onion, as observed by GFP expression, was essentially constitutive, and profiles of regulation were typical of those observed in dicotyledonous plants. Inhibition of CaMV35s regulated gene expression was only observed in one transformant. Both the expression of GFP and tolerance to phosphinothricin appeared to be inherited in a Mendelian fashion. Levels of expression in F1 offspring varied, presumably due to environmental and genetic factors. However, it appeared that copy number did strongly influence GFP protein production and expression. In the majority of plants there were no obvious detrimental phenotypic effects caused by the transgene, the integration event, or Somaclonal variation due to the need to perform tissue culture.     

A calcium-binding sialoglycoprotein associated with an apparent eggshell membrane of Globodera rostochiensis

Energy dispersive X-ray microanalysis has been used to study the binding of two inhibitors of hatching in Globodera rostochiensis, ruthenium red and lanthanum, to a calcium binding site on the eggshell. Eggs were treated with either buffered ruthenium red or buffer only, and the levels of ruthenium and calcium that could be detected from the surface of eggshells were measured before and after repeated etching in a plasma oven. Counts for ruthenium increased during etching to a maximum after 8–12 min, just before the eggshell was eroded from the juvenile. The calcium levels on eggshells binding ruthenium red were suppressed relative to those from buffer. Similar experiments with lanthanum showed a significant increase in this inhibitor after 10 min etching and an accompanying suppression of calcium levels. The nature of the binding site was probed by measuring ruthenium red binding after pretreatment with selected enzymes. A non-specific protease, pronase, reduced the subsequent binding of this inhibitor but the site was resistant to trypsin in the presence of calcium. Neuraminidase also had an effect suggesting that the protein is a sialoglycoprotein. Phospholipase A2 but not Phospholipase C influenced the subsequent binding of ruthenium red. Phospholipase A₂ caused a loss of hatching ability and the degradation of a phospholipid from the eggshell. Ruthenium red binding was also reduced by treatment of eggshells with detergents or sonication. Apparently a calcium-binding sialoglycoprotein forms part of the integral proteins of an eggshell membrane located on the inner surface of the eggshell.     

Leaf phenology mediates provenance differences in herbivore populations on valley oaks in a common garden

1. Plants from different populations often display a variation in herbivore resistance. However, it is rarely understood what plant traits mediate such differences. 2. It was tested how leaf phenology affects herbivore populations in a 15‐year‐old common garden of valley oaks (Quercus lobata Née) with different populations and maternal parents from throughout the Q. lobata range. 3. The abundance of leaf miners (Stigmella sp. Shrank) and leaf phenology of oaks in the common garden was measured. 4. Leaf miner abundance varied among provenance locations (population), but not among maternal parents within populations. Leaf phenology varied by provenance location and maternal parent, and trees that leafed out earlier accrued higher leaf‐miner abundance. Path analysis indicated that leaf phenology was the likely driver of provenance and parental differences in resistance to leaf miners. 5. Understanding population differences is particularly important when considering transport of genotypes for ornamental or restoration purposes. The present study suggests that similarity in leaf phenology may be one factor that could be used to find genotypes with a similar herbivore resistance to local genotypes.     

Field experiments on flow refugia in streams

1. Field experiments were carried out to determine whether animals move into areas of low shear stress during periods of peak flow and therefore avoid hydraulic disturbance.
2. Flow at the scale of 0.05 m² patches was reduced experimentally by creating artificial 'refugia'. Invertebrate colonization of cages with 1.1 mm mesh sides, which provided such potential refugia by reducing hydraulic forces within the cage, was compared with colonization of cages with coarser (15 mm) mesh which did not restrict the flow.
3. Colonization of these cages was tested over a series of weekly periods in two different streams. Nine trials were completed in a stream with abundant natural flow refugia (Broadstone Stream, SE England), and during three of these trials strong spates occurred. Six trials were completed in a stream with comparatively few natural flow refugia (Dargall Lane, SW Scotland), and peak flows were relatively less.
4. In Broadstone Stream, the relative colonization of refugium cages was greatest during periods of peak flow, suggesting animals had used these low-flow areas as refugia during spates. Use of the artificial refugia did not occur in Dargall Lane, at least at the flows achieved during the trials.
5. Our results are consistent with the hypothesis that stream invertebrates accumulate in refugia during high flow disturbances. Whether their distribution among patches is altered by active or passive means remains unclear.     

A NOTE ON THE USE OF ROGOSA AND MODIFIED ROGOSA MEDIA FOR THE ENUMERATION AND CULTURE OF LACTOBACILLI

SUMMARY: The modified Rogosa media (acetate broth and acetate agar) developed by Mabbitt & Zielinska (1956) have been compared with tomato juice media for the growth of representative Lactobacillus species.
Most of the species tested grew markedly better in acetate broth than in tomato dextrose broth, but some cultures did not grow well in the acetate broth although they grew normally in tomato dextrose broth. The acetate agar was satisfactory for counting those lactobacilli cultured at 30° but was inferior to tomato dextrose agar for counting those grown at 37°.     

VARIABILITY IN VEGETATION EFFECTS ON DENSITY AND NESTING SUCCESS OF GRASSLAND BIRDS

Abstract: The structure of vegetation in grassland systems, unlike that in forest systems, varies dramatically among years on the same sites, and among regions with similar vegetation. The role of this variation in vegetation structure on bird density and nesting success of grassland birds is poorly understood, primarily because few studies have included sufficiently large temporal and spatial scales to capture the variation in vegetation structure, bird density, or nesting success. To date, no large-scale study on grassland birds has been conducted to investigate whether grassland bird density and nesting success respond similarly to changes in vegetation structure. However, reliable management recommendations require investigations into the distribution and nesting success of grassland birds over larger temporal and spatial scales. In addition, studies need to examine whether bird density and nesting success respond similarly to changing environmental conditions. We investigated the effect of vegetation structure on the density and nesting success of 3 grassland-nesting birds: clay-colored sparrow (Spizella pallida), Savannah sparrow (Passerculus sandwichensis), and bobolink (Dolichonyx oryzivorus) in 3 regions of the northern tallgrass prairie in 1998–2001. Few vegetation features influenced the densities of our study species, and each species responded differently to those vegetation variables. We could identify only 1 variable that clearly influenced nesting success of 1 species: clay-colored sparrow nesting success increased with increasing percentage of nest cover from the surrounding vegetation. Because responses of avian density and nesting success to vegetation measures varied among regions, years, and species, land managers at all times need to provide grasslands with different types of vegetation structure. Management guidelines developed from small-scale, short-term studies may lead to misrepresentations of the needs of grassland-nesting birds.     

Ultrastructural Effects of Lactoferrin Binding on Giardia lamblia Trophozoites

ABSTRACT. Lactoferrin and its derived N-terminal peptide may be important host defenses against Giardia lamblia . We showed earlier that lactoferrin and the derived peptides have potent giardicidal activity in vitro. Using indirect immunofluorescence, we now demonstrate binding of lactoferrin and the peptides to the live trophozoite surface. Iron strongly inhibited binding of lactoferrin, and decreased binding of the peptides, while certain divalent metal ions decreased binding of all forms by about half. Lactoferrin and the peptedes caused striking and complex morphologic changes in the trophozoite plasmalemma, endomembranes and cytoskeleton, and increases the electron density of the lysosome-like peripheral vacuoles.