Optimisation of introducing foreign genes into egg cells and zygotes of wheat (Triticum aestivum L.) via microinjection

Summary An expeditious and highly efficient technique of microinjection has been developed with the aim of introducing exogenous DNA into egg cells and zygotes of wheat. Using a mechanical-dissection method and a novel immobilisation approach enabled us to microinject around 15 egg cells of wheat per hour. Exposing the protoplasts to a high-frequency alternating-current field for immobilisation, a significantly higher transient expression rate of the injected genes (46% and 52% for egg cells and zygotes, respectively) could be achieved than reported thus far for plant protoplasts. Whether this high transformation efficiency is due to the highfrequency electrical field applied for immobilising the protoplasts is not known. The transformation rate appeared to be a factor depending upon the time of egg cell isolation. According to the ultrastructural observations this seems to reflect a variation in competence of the egg cells during in situ development. In order to conduct studies directed towards establishing the optimal timewindow for DNA delivery into the fertilised egg cell, the time course of DNA dynamics during zygotic development has been quantified via quantitative microspectrofluorometry.Abbreviations AC alternating current - DAE days after emasculation - FDA fluorescein diacetate - HAP hours after pollination     

生态场理论在沙地云杉种群更新中的应用

生态场理论是物理生态学的基本理论,它基于植物分布格局,植物生长模型,植物竞争以及竞争利用的研究,研究森林草原过滤带上沙地云杉种群在更新中产生的生态场的特性,结果表明,沙地云杉种群生态场是半椭圆形特殊的非均匀场,生态场强度随着母树距离的增大而变化,开始时迅速增大,３￣５ｍ处场强达到最大,然后逐渐下降,由于生态场的重叠而使叠加部分的场强明显增大,生态场的作用范围与坡度,坡长等地形因子及树高,冠半径等测     

Behavioral observations of codling moth, Cydia pomonella, in orchards permeated with synthetic pheromone

Mating disruption of codling moth, Cydia pomonella, was studied in apple orchards treated with the main pheromone compound codlemone, (E,E)-8,10-dodecadienol, and a blend of codlemone and codlemone acetate, (E,E)-8,10-dodecadienyl acetate, a strong pheromone antagonist. Codlemone alone and the pheromone/antagonist-blend had a similar effect on the behavior of males emerging into air-permeated orchards. Male flights within tree canopy and upwind orientation along tree rows were strongly enhanced by both formulations, compared to untreated plots. However, the codlemone/codlemone acetate-blend increased the rate of cross-wind and downwind flights within the orchard, compared to codlemone alone. The major difference between these two formulations was that males from nearby, untreated orchards were attracted towards orchards treated with codlemone, but not towards treatments with codlemone/codlemone acetate. Additional tests were done with an equilibrium blend of codlemone and its geometric isomers. Aerial pheromone concentrations in the orchards were recorded by the field electroantennogram technique. Decreasing pheromone concentrations towards the upper part of the tree canopy, together with the stimulation of male flight activity by synthetic pheromone explains failures to control codling moth at high population densities with current dispenser formulations.     

Measuring Daphnia life history in the wild: The efficacy of individual field cages

Life‐history studies are often conducted in a laboratory environment where it is easy to assay individual animals. However, factors such as temperature, photoperiod, and nutrition vary greatly between laboratory and field environments, making it difficult to compare results. Consequently, there is a need to study individual life histories in the field, but this is currently difficult in systems such as Daphnia where it is not possible to mark and track individual animals. Here, we present a proof of principle study showing that field cages are a reliable method for collecting individual‐level life‐history data in Daphnia magna. As a first step, we compared the life history of paired animals reared outside and inside cages to test the hypothesis that cages allow free flow of algal food resources. We then used a seminatural mesocosm setting to compare the performance of individual field cages versus glass jars refilled with mesocosm water each day. We found that cages did not inhibit food flow and that differences in life histories between three clones detected in the jar assays were also detectable using the much less labor‐intensive field cages. We conclude that field cages are a feasible approach for collecting individual‐level life‐history data in systems such as Daphnia where individual animals cannot be marked and tracked.     

Effects of enhanced productivity of resources shared by predators in a food‐web module: Comparing results of a field experiment to predictions of mathematical models of intra‐guild predation

We compared the response to resource enhancement of a simple empirical model of intra‐guild predation (IGP) to the predictions of published, simple mathematical models of asymmetric IGP (a generalist IG Predator that feeds both on a specialist IG Prey and a Resource that it shares with the IG Prey). The empirical model was a food‐web module created by pooling species abundances across many families in a speciose community of soil micro‐arthropods into three categories: IG Predator (large predatory mites), IG Prey (small predatory mites), and a shared Resource (fungivorous mites and springtails). By pooling abundances of species belonging to broadly defined functional groups, we tested the hypothesis that IGP is a dominant organizing principle in this community. Simple mathematical models of asymmetric IGP predict that increased input of nutrients and energy to the shared Resource will increase the equilibrium density of Resource and IG Predator, but will decrease that of IG Prey. In a field experiment, we observed how the three categories of the empirical model responded to two rates of addition of artificial detritus, which enhanced the food of fungivores, the Resource of the IGP module. By the experiment''s end, fungivore densities had increased ~1.5× (ratio of pooled fungivore densities in the higher‐input treatment to plots with no addition of detritus), and densities of IG Predators had increased ~4×. Contrary to the prediction of mathematical models, IG Prey had not decreased, but instead had increased ~1.5×. We discuss possible reasons for the failure of the empirical model to agree with IGP theory. We then explore analogies between the behavior of the empirical model and another mathematical model of trophic interactions as one way to gain insights into the trophic connections in this community. We also propose one way forward for reporting comparisons of simple empirical and mathematical models.     

Cell density dependent response of E. Coli cells to weak ELF magnetic fields

The effects of weak magnetic fields of extremely low frequency (ELF) on E. coli K12 AB1157 cells were studied by the method of anomalous viscosity time dependencies (AVTD). E. coli cells at different densities within a range of 5 × 10⁵–10⁹ cell/ml were exposed to ELF (sinusoidal, 30 μT peak, 15 min) at a frequency of 9 Hz. A transient effect with maximum 40–120 min after exposure was observed. Kinetics of the per-cell-normalised ELF effects fitted well to a Gaussian distribution for all densities during exposure. A maximum value of these kinetics and a time for this maximum were strongly dependent on the cell density during exposure. These data suggest a cell-to-cell interaction during response to ELF. Both dependencies had three regions close to a plateau within the ranges of 3 × 10⁵ − 2 × 10⁷ cell/ml, 4 × 10⁷ − 2 × 10⁸ cell/ml and 4 × 10⁸–10⁹ cell/ml and two rather sharp transitions between these plateaus. The effect reached a maximum value at a density of 4 × 10⁸ cell/ml. Practically no effect was observed at the lowest density of 3 × 10⁵ cell/ml. The data suggested that the ELF effect was mainly caused by a secondary rather than a primary reaction. The filtrates from exposed cells neither induced significant AVTD changes in unexposed cells nor increased the ELF effect when were added to cells before exposure. The data did not provide evidence for significant contribution of stable chemical messengers, but some unstable compounds such as radicals could be involved in the mechanism of cell-to-cell interaction during response to ELF. The results obtained were also in accordance with a model based on an re-emission of secondary photons during resonance fluorescence. Bioelectromagnetics 19:300–309, 1998. © 1998 Wiley-Liss, Inc.     

Electric field-induced changes in agonist-stimulated calcium fluxes of human HL-60 leukemia cells

The mechanism of biological effects of extremely-low-frequency electric and magnetic fields may involve induced changes of Ca²⁺ transport through plasma membrane ion channels. In this study we investigated the effects of externally applied, low-intensity 60 Hz electric (E) fields (0.5 V/m, current density 0.8 A/m²⁺) on the agonist-induced Ca²⁺ fluxes of HL-60 leukemia cells. The suspensions of HL-60 cells received E-field or sham exposure for 60 min and were simultaneously stimulated either by 1 μM ATP or by 100 μM histamine or were not stimulated at all. After E-field or sham exposure, the responses of the intracellular calcium levels of the cells to different concentrations of ATP (0.2–100 μM) were assessed. Compared with control cells, exposure of ATP-activated cells to an E-field resulted in a 20–30% decrease in the magnitude of [Ca²⁺]_i elevation induced by a low concentration of ATP (<1 μM). In contrast, exposure of histamine-activated HL-60 cells resulted in a 20–40% increase of ATP-induced elevation of [Ca²⁺]_i. E-field exposure had no effect on non-activated cells. Kinetic analysis of concentration-response plots also showed that compared with control cells, exposure to the E-field resulted in increases of the Michaelis constant, K_m, value in ATP-treated cells and of the maximal [Ca²⁺]_i peak rise in histamine-treated HL-60 cells. The observed effects were reversible, indicating the absence of permanent structural damages induced by acute 60 min exposure to electric fields. These results demonstrate that low-intensity electric fields can alter calcium distribution in cells, most probably due to the effect on receptor-operated Ca²⁺ and/or ion channels. Bioelectromagnetics 19:366–376, 1998. © 1998 Wiley-Liss, Inc.     

Mechanism of action of weak electromagnetic field on ionic currents in aqueous solutions of amino acids

To analyze the mechanisms of action of weak electric and magnetic fields on ion motion in solution, a new approach is outlined allowing for collective interactions of large ionic ensembles with the external field. Bioelectromagnetics 18:25–27, 1997. © 1997 Wiley-Liss, Inc.     

Applied DC magnetic fields cause alterations in the time of cell divisions and developmental abnormalities in early sea urchin embryos

Most work on magnetic field effects focuses on AC fields. The present study demonstrates that exposure to medium-strength (10 mT-0.1 T) static magnetic fields can alter the early embryonic development of two species of sea urchin embryos. Batches of fertilized eggs from two species of urchin were exposed to fields produced by permanent magnets. Samples of the continuous cultures were scored for the timing of the first two cell divisions, time of hatching, and incidence of exogastrulation. It was found that static fields delay the onset of mitosis in both species by an amount dependent on the exposure timing relative to fertilization. The exposure time that caused the maximum effect differed between the two species. Thirty millitesla fields, but not 15 mT fields, caused an eightfold increase in the incidence of exogastrulation in Lytechinus pictus, whereas neither of these fields produced exogastrulation in Strongylocentrotus purpuratus. Bioelectromagnetics 18:255–263, 1997. © 1997 Wiley-Liss, Inc.