Nickel and Cadmium-related Changes in Growth, Plasma Membrane Lipid Composition, ATPase Hydrolytic Activity and Proton-pumping of Rice (Oryza sativa L. cv. Bahia) Shoots

Six-day-old rice plants (Oryza sativa L. cv. Bahia) were grownin the presence of 0.5 mol m^–3 Ni or Cd for 5 or 10 d.Five days after treatment, some plants were transferred to amedium containing no heavy metal for 5 or 10 d. Plasmalemmavesicles from shoots of treated, transferred (recovery experiments)and control plants were isolated, ATPase activity and proton-pumpingwere assessed, and free sterols and phospholipids determined.The ATPase hydrolytic activity was increased by 37% and 34%in 5 and 10 d Cd-treated plants, respectively; and by 66% in5 and 10 d Ni-treated plants. However, neither the initial rateof H⁺ transport nor the proton-pumping rate at steady-statewere significantly affected by the treatments. The relativeproportion (%) of the plasmalemma sterols campesterol and ⁵-avenasteroldecreased while sitosterol increased during heavy metal treatment.The overall plasmalemma phospholipid fatty acyl chain lengthand degree of unsaturation were also reduced. In experimentswhere plants recovered from Cd and Ni treatment, differencesbetween treated and control plants were reverted, particularlyin 10 d Ni-recovered plants. The possible involvement of lipidsin the regulation of the plasmalemma ATPase as well as the relationshipbetween growth, ATPase and adaptations to stress conditionsare discussed. Key words: Cadmium, nickel, sterols, phospholipids, ATPase     

Genetic structure and population dynamics of a heteroecious plant pathogen Melampsora larici-epitea in short-rotation coppice willow plantations

Complex life strategies are common among plant pathogens belonging to rust fungi ( Uredinales ). The heteroecious willow rust Melampsora larici-epitea produces five spore stages and alternates on larch ( Larix ). To shed light on the epidemiology of this pathogen, amplified fragment length polymorphisms (AFLPs) were used to determine the genetic diversity and genetic structure of rust samples collected from coppice willow ( Salix ) plantations at three UK sites (LA, CA and MC) over three sampling dates (September 2000, July 2001 and September 2001). Of the total of 819 isolates, 465 were unique AFLP phenotypes and there was a shift in genotype diversity between the two seasons (0.67 in 2000 and 0.87–0.89 in 2001). No phenotypes were common between the two seasons within a site, suggesting that the rust did not overwinter as an asexual stage within plantations. A temporal analysis detected large amounts of genetic drift ( F _S = 0.15–0.26) between the two seasons and very small effective population sizes ( N _e  =  2–3) within sites. These results all point to a new colonization of the plantations by the rust in the second season (2001). The F _ST-analogue values were Φ_CT = 0.121, Weir and Cockerham's θ = 0.086 and the Bayesian estimate θ^B = 0.087–0.096. The results suggest that the sources of inoculum were somewhat localized and the same sources were mainly responsible for disease epidemics in LA and CA over the two seasons. The relatively low F _ST-values among sites (0.055–0.13) suggest the existence of significant gene flow among the three sampled sites.     

FLUORESCENT MEASUREMENTS OF DNA, RNA AND PROTEINS TO PERFORM COMPARATIVE ANALYSES OF MICROBIAL COMMUNITIES FROM THE ENVIRONMENTS

Simple and rapid methods for the quantification of DNA, RNA and proteins using specific fluorescent dyes are proposed for the comparison and monitoring of microbial communities from the environment. The purpose of this study was the use of straightforward in situ methods which voided the need for preservation of samples and the risk of potential degradation and quantitative changes during transportation. Aside from this, methods used to obtain information on environmental microbial communities are generally time-consuming and present certain difficulty above all when working on solid substrates such as soils and rocks. New generation fluorescent dyes that bind specifically to DNA, RNA and proteins allow simple and rapid estimates of these biomolecules in crude environmental samples.

PRACTICAL APPLICATIONS

Monitoring the metabolic state of microbial communities on different substrates and environments is a requirement for comparing samples and assessing the participation of microorganisms in a variety of processes. Solid substrates are not easily analyzed by microscopic techniques and they require long processing times and tedious work. Aside from this, only a minor fraction (<1%) of microorganisms in most natural environments can be cultured in standard microbiological media ( Ward et al. 1990 ). Other studies using incorporation of labeled substrates to approach activity rates or biomolecule extractions represent complex and long procedures during environmental studies.In order to evaluate microbial communities in a variety of substrates and environments, rapid and simple methods are proposed by measuring DNA, RNA and/or proteins using specific fluorescent dyes, without a need for prior purification, from crude solid samples.     

Biosynthetic pathways of the pheromone of the Egyptian armyworm Spodoptera littoralis

Abstract Most insect pheromones comprise multicomponent blends of geometric or optical isomers, and one major question is how insects produce species‐specific ratios of components for successful reproductive isolation. Key enzymes suggested to be involved in pheromone biosynthesis are acetyl‐coenzyme A carboxylase and fatty acyl synthetase, chain‐shortening enzymes, desaturases, elongases, reductases, oxidases, and alcohol acetyl transferases. The female pheromone composition of the Egyptian armyworm Spodoptera littoralis (Boisd.) is highly dependent on the origin of the strain. In this review, we present a summary of the different reported pheromone compositions of the moth, including from our recent studies on this subject, as well as the biosynthetic routes to the different components and the molecular approaches involved. In addition, the key role played in the proposed biosynthetic pathways by a number of important biosynthetic enzymes, such as chain shortening enzymes, desaturases and alcohol acetyl transferases, is outlined, as well as the latest developments on the inhibition of these enzymes.     

Effect of thermal acclimation on preferred temperatures in two mygalomorph spiders inhabiting contrasting habitats

Variations in the preferred temperatures during the rest periods of Grammostola rosea Walckenaer and Paraphysa parvula Pocock, two mygalomorph spiders occupying different habitats in central Chile, are analyzed. The former inhabits arid and semi‐arid lowland near plant communities, composed of shrubs (evergreens with small leathery leaves) and small trees; the latter is found in the central mountains of the Chilean Andes, above 2000 m.a.s.l. The preferred temperatures of these spiders at different times of day and exposure to cold (15 °C) and warm (25 °C) acclimation temperatures are compared. Body mass does not affect the preferred temperature of the larger spider G. rosea, although P. parvula, a spider with half of the body mass of G. rosea, shows a decrease in preferred temperature with body mass. This can be explained by a higher plasticity and thermal sensitivity of the smaller species as result of increased surface : volume ratio. The preferred temperature increases with the hour of the day under both acclimation conditions in P. parvula and in cold‐acclimated G. rosea, which is likely associated with crepuscular and nocturnal behaviour in both species. Grammostola rosea shows temperature preferences lower than those of P. parvula under both acclimation conditions. The increase of the acclimation temperature from 15 to 25 °C results in an increment of 2–3 °C in the preferred temperature of P. parvula but only 0.2 °C in that of G. rosea. Two contrasting lifestyle strategies are found: a small mygalomorph spider with phenotypic plasticity and adaptation to the fluctuating environment of high altitude, and a large mygalomorph spider with higher thermal inertia adapted to the more stable environment of lowlands.     

In Vitro Culture of Phytomonas Sp. Isolated from Euphorbia characias. Metabolic Studies by 1H NMR

ABSTRACT. We describe the in vitro culture of Phytomonas species isolated from Euphorbia characias . The best choice between tested media was SDM-79, in which promastigotes, after 6 days of culture, reached cell densities as high as 4 × 10⁷ cells/ml. Cells growing in LIT or MTL medium showed longer division times and lower cell densities. We succeeded in obtaining Phytomonas sp. amastigote and spheromastigote forms in modified GRACE's medium, yielding transformation rates of up to 70%. Electron microscopy studies were performed in order to characterize the ultrastructural features of these forms obtained in vitro. On the other hand, metabolic studies based on qualitative (nuclear magnetic resonance spectroscopy) and quantitative metabolic methods (enzymatic assays) showed that promastigote forms secreted mainly ethanol, acetate, glycine, glycerol, piruvate and succinate in SDM-79 medium, whereas the major metabolites found after transformation in modified Grace's medium were ethanol, acetate, glycine, piruvate and smaller amounts of glycerol.     

The Different Behavior of Diphtheria Toxin, Modeccin and Ricin in HeLa Cells Infected with Trypanosoma cruzi

ABSTRACT. We have studied the action of diphtheria toxin, modeccin and ricin on HeLa cells infected by Trypanosoma cruzi . Parasitized HeLa cells were resistant to diphtheria toxin and modeccin, whereas non-parasitized cells from the same cultures and control cultures showed cytopathological alterations. Protein synthesis, assayed by the incorporation of labelled methionine, diminished in toxin-treated control cultures but remained unaltered in the infected ones, compared to synthesis by untreated infected cells. Ricin, on the other hand, is a toxin that enters the cytoplasm by endocytosis. It has greater cytopathological effects in parasitized cells than in non-parasitized ones from the same cultures or uninfected control cells. Protein synthesis was inhibited in infected cultures treated with ricin.