Phloem cytochemical modification and gene expression following the recovery of apple plants from apple proliferation disease.

Recovery of apple trees from apple proliferation was studied by combining ultrastructural, cytochemical, and gene expression analyses to possibly reveal changes linked to recovery-associated resistance. When compared with either healthy or visibly diseased plants, recovered apple trees showed abnormal callose and phloem-protein accumulation in their leaf phloem. Although cytochemical localization detected Ca(2+) ions in the phloem of all the three plant groups, Ca(2+) concentration was remarkably higher in the phloem cytosol of recovered trees. The expression patterns of five genes encoding callose synthase and of four genes encoding phloem proteins were analyzed by quantitative real-time reverse transcription-polymerase chain reaction. In comparison to both healthy and diseased plants, four of the above nine genes were remarkably up-regulated in recovered trees. As in infected apple trees, phytoplasma disappear from the crown during winter, but persist in the roots, and it is suggested that callose synthesis/deposition and phloem-protein plugging of the sieve tubes would form physical barriers preventing the recolonization of the crown during the following spring. Since callose deposition and phloem-protein aggregation are both Ca(2+)-dependent processes, the present results suggest that an inward flux of Ca(2+) across the phloem plasma membrane could act as a signal for activating defense reactions leading to recovery in phytoplasma-infected apple trees.

Measuring, modelling and testing ozone exposure, flux and effects on vegetation in southern European conditions--what does not work? A review from Italy.

Ozone (O3) exposure at Italian background sites exceeds UN/ECE concentration-based critical levels (CLe(c)), if expressed in terms of AOT40. Yet the occurrence of adverse effects of O3 on forests and crops is controversial. Possible reasons include (i) ability of response indicators to provide an unbiased estimate of O3 effects, (ii) setting of current CLe(c) in terms of cut-off value and accumulation level, (iii) response functions adopted to infer a critical level, (iv) environmental limitation to O3 uptake and (v) inherent characteristics of Mediterranean vegetation. In particular, the two latter points suggest that critical levels based on accumulated stomatal flux (CLe(f)) can be a better predictor of O3 risk than CLe(c). While this concept is largely acknowledged, a number of factors may limit its applicability for routine monitoring. This paper reviews levels, uptake and vegetation response to O3 in Italy over recent years to discuss value, uncertainty and feasibility of different approaches to risk assessment.

The inhibitory effects of coumarin on the germination of durum wheat (Triticum turgidum ssp. durum, cv. Simeto) seeds.

The event chain leading to germination blockage in durum wheat (Triticum turgidum ssp. durum Desf.) seeds exposed to the allelochemical coumarin (2H-chromen-2-one) was studied. The physiological and biochemical aspects thought to be critical for a successful seed germination were measured. At concentrations above 200 microM: , coumarin inhibited seed germination in a concentration-dependent manner. Inhibition occurred early during seed imbibition (phase I), was rapid, and irreversible. During phase I, coumarin inhibited water uptake, electrolyte retention capacity, and O(2) consumption. Later on, coumarin delayed the reactivation of peroxidases, enhanced the activity of superoxide dismutase, decreased the activities of selected marker enzymes for metabolic resumption, and repressed the transcription of molecular chaperones involved in secretory pathways. Insufficient and/or late seed rehydration caused by coumarin could have delayed membrane stabilization or decreased respiratory O(2) consumption, both of which are conducive to an overproduction of reactive O(2) species. Being unbalanced by an adequate upsurge of antioxidant defense systems, the resulting oxidative stress might have ultimately interfered with the germination program.

Foliar antioxidant status of adult Mediterranean oak species (Quercus ilex L. and Q. pubescens Willd.) exposed to permanent CO2-enrichment and to seasonal water stress.

Foliar antioxidants were measured in adult individuals of holm oak (Quercus ilex L.) and white oak (Q. pubescens Willd.) growing in the field either within the vicinity of natural CO2 springs or at a nearby control site under ambient CO2, which had been previously exposed to either daily irrigation or no irrigation. In oak trees permanently exposed to elevated CO2 the activities of antioxidant enzymes tended to be lower and the ascorbate pool was larger and more in reduced form, suggesting an attenuation of the oxidative risk in the CO2-enriched trees. In the enriched individuals of both species. the imposition of water shortage significantly increased the size of the glutathione pool and the total superoxide dismutase activity in a species-specific manner. Moreover, water-stressed trees exposed to elevated CO2 tended to have higher catalase and ascorbate peroxidase activities than water-stressed control trees. Such changes may reflect the need for an enhanced compensatory effort when trees acclimated to elevated CO2 are exposed to oxidative stress-promoting environmental factors, such as water shortage.

Soluble peroxidase from winter wheat seedlings with phenoloxidase-like activity.

Peroxidase isozymes from winter wheat (Triticum aestivum L. cv Orso) seedlings extracts showed phenoloxidase-like activity, becoming visible on polyacrylamide gels also in the absence of hydrogen peroxide. The results obtained after a characterization of the two activities, based on their substrate specificity, on their selective inhibition, and on the possible occurrence of artifacts, suggested the existence of polyfunctional peroxidase isozymes. Different isozymes possessing only phenol oxidase activity were not found in the same plant material. This appears to be the first evidence of phenoloxidase-acting isoperoxidases in winter wheat.

Laccase assay by means of high-performance liquid chromatography.

Spectrophotometric determination of laccase activity may be affected by the formation of quinoid chromophores arising from nonenzymatic oxidations interfering with enzymatic reactions. Km values for guaiacol obtained by spectrophotometric and HPLC methods confirm the above hypothesis. HPLC results are particularly useful for the assay of laccase activity on natural phenolic extracts.