Wounded leaves of Arabidopsis thaliana show transient immunity to Botrytis cinerea, the causal agent of grey mould. Using a fluorescent probe, histological staining and a luminol assay, we now show that reactive oxygen species (ROS), including H2O2 and O2
−, are produced within minutes after wounding. ROS are formed in the absence of the enzymes Atrboh D and F and can be prevented by diphenylene iodonium (DPI) or catalase. H2O2 was shown to protect plants upon exogenous application. ROS accumulation and resistance to B. cinerea were abolished when wounded leaves were incubated under dry conditions, an effect that was found to depend on abscisic acid (ABA). Accordingly, ABA biosynthesis mutants (aba2 and aba3) were still fully resistant under dry conditions even without wounding. Under dry conditions, wounded plants contained higher ABA levels and displayed enhanced expression of ABA-dependent and ABA-reporter genes. Mutants impaired in cutin synthesis such as bdg and lacs2.3 are already known to display a high level of resistance to B. cinerea and were found to produce ROS even when leaves were not wounded. An increased permeability of the cuticle and enhanced ROS production were detected in aba2 and aba3 mutants as described for bdg and lacs2.3. Moreover, leaf surfaces treated with cutinase produced ROS and became more protected to B. cinerea. Thus, increased permeability of the cuticle is strongly linked with ROS formation and resistance to B. cinerea. The amount of oxalic acid, an inhibitor of ROS secreted by B. cinerea could be reduced using plants over expressing a fungal oxalate decarboxylase of Trametes versicolor. Infection of such plants resulted in a faster ROS accumulation and resistance to B. cinerea than that observed in untransformed controls, demonstrating the importance of fungal suppression of ROS formation by oxalic acid. Thus, changes in the diffusive properties of the cuticle are linked with the induction ROS and attending innate defenses.
This paper compares different types of simulated draws over a range of number of draws in generating Bayesian efficient designs for stated choice (SC) studies. The paper examines how closely pseudo Monte Carlo, quasi Monte Carlo and Gaussian quadrature methods are able to replicate the true levels of Bayesian efficiency for SC designs of various dimensions. The authors conclude that the predominantly employed method of using pseudo Monte Carlo draws is unlikely to result in leading to truly Bayesian efficient SC designs. The quasi Monte Carlo methods analyzed here (Halton, Sobol, and Modified Latin Hypercube Sampling) all clearly outperform the pseudo Monte Carlo draws. However, the Gaussian quadrature method examined in this paper, incremental Gaussian quadrature, outperforms all, and is therefore the recommended approximation method for the calculation of Bayesian efficiency of stated choice designs.
Red-light cameras are effective in reducing total casualty crashes. The evidence is less conclusive on total collisions, specific casualty collision types and violations, where reductions achieved could be explained by the play of chance. Most evaluations did not adjust for RTM or spillover, affecting their accuracy. Larger and better controlled studies are needed.
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