Season-long exposure of bilberry plants to realistic and future ozone pollution improves the nutraceutical quality of fruits.

Ozone (O3) is a phytotoxic air pollutant capable of limiting plant yield and growth, and altering the quality of edible plant products. This study aimed to investigate the effects of long-term O3 exposure at realistic and future concentrations (applied during fruit development) not only on morphological, physiological, and biochemical plant/leaf traits of Vaccinium myrtillus but also on its fruit yield and quality. Three-year-old saplings were grown from May to July under three levels of O3 concentration [1.0, 1.5 and 2.0 times the ambient air concentrations, denoted as AA, 1.5_AA and 2.0_AA], using a new-generation O3 Free Air Controlled Exposure system. Ozone induced oxidative pressure and membrane denaturation as confirmed by the accumulation of anion superoxide, hydrogen peroxide (•O2-: +39 and + 29%; H2O2: +55 and + 59% in 1.5_AA and 2.0_AA, respectively, compared with AA), and malondialdehyde by-product (1.4- and 2.5-fold higher than AA, in 1.5_AA and 2.0_AA, respectively). The observed oxidative burst likely affected several cellular structures interested by photosynthetic processes (e.g., decrease of the maximum rate of carboxylation: -30%). This constraint likely induced a decline in plant vitality and a different partitioning of biomass allocation between above and below organs. An accelerated maturation of bilberries due to O3 was reported, suggesting that plants grown under harsher environmental conditions suffered from metabolic changes associated with early ripening. Increasing O3 concentrations might be responsible for an alteration of the ratio between oxidation and reduction processes mechanisms that was followed by a loss of integrity of membranes, so limiting the availability of energy/resources, triggering enzymatic oxidation of phenols to red/purple pigments, and promoting fruit maturation. To the best of our knowledge, this is the first research showing that long-term O3 exposure during bilberry fruit development influenced not only several plant/leaf traits, but also fruit nutraceutical quality at the time of harvest.

Red versus green leaves: transcriptomic comparison of foliar senescence between two Prunus cerasifera genotypes.

The final stage of leaf ontogenesis is represented by senescence, a highly regulated process driven by a sequential cellular breakdown involving, as the first step, chloroplast dismantling with consequent reduction of photosynthetic efficiency. Different processes, such as pigment accumulation, could protect the vulnerable photosynthetic apparatus of senescent leaves. Although several studies have produced transcriptomic data on foliar senescence, just few works have attempted to explain differences in red and green leaves throughout ontogenesis. In this work, a transcriptomic approach was used on green and red leaves of Prunus cerasifera to unveil molecular differences from leaf maturity to senescence. Our analysis revealed a higher gene regulation in red leaves compared to green ones, during leaf transition. Most of the observed DEGs were shared and involved in transcription factor activities, senescing processes and cell wall remodelling. Significant differences were detected in cellular functions: genes related to photosystem I and II were highly down-regulated in the green genotype, whereas transcripts involved in flavonoid biosynthesis, such as UDP glucose-flavonoid-3-O-glucosyltransferase (UFGT) were exclusively up-regulated in red leaves. In addition, cellular functions involved in stress response (glutathione-S-transferase, Pathogen-Related) and sugar metabolism, such as three threalose-6-phosphate synthases, were activated in senescent red leaves. In conclusion, data suggests that P. cerasifera red genotypes can regulate a set of genes and molecular mechanisms that cope with senescence, promoting more advantages during leaf ontogenesis than compared to the green ones.

Can nutrient fertilization mitigate the effects of ozone exposure on an ozone-sensitive poplar clone?

We tested the independent and interactive effects of nitrogen (N; 0 and 80 kg ha-1), phosphorus (P; 0, 40 and 80 kg ha-1), and ozone (O3) application/exposure [ambient concentration (AA), 1.5 × AA and 2.0 × AA] for five consecutive months on biochemical traits of the O3-sensitive Oxford poplar clone. Plants exposed to O3 showed visible injury and an alteration of membrane integrity, as confirmed by the malondialdehyde by-product accumulation (+3 and +17% under 1.5 × AA and 2.0 × AA conditions, in comparison to AA). This was probably due to O3-induced oxidative damage, as documented by the production of superoxide anion radical (O2-, +27 and +63%, respectively). Ozone per se, independently from the concentrations, induced multiple signals (e.g., alteration of cellular redox state, increase of abscisic acid/indole-3-acetic acid ratio and reduction of proline content) that might be part of premature leaf senescence processes. By contrast, nutrient fertilization (both N and P) reduced reactive oxygen species accumulation (as confirmed by the decreased O2- and hydrogen peroxide content), resulting in enhanced membrane stability. This was probably due to the simultaneous involvement of antioxidant compounds (e.g., carotenoids, ascorbate and glutathione) and osmoprotectants (e.g., proline) that regulate the detoxification processes of coping with oxidative stress by reducing the O3 sensitivity of Oxford clone. These mitigation effects were effective only under AA and 1.5 × AA conditions. Nitrogen and P supply activated a free radical scavenging system that was not able to delay leaf senescence and mitigate the adverse effects of a general peroxidation due to the highest O3 concentrations.

Impact of mechanical mowing and chemical treatment on phytosociological, pedochemical and biological parameters in roadside soils and vegetation.

Many chemical and non-chemical strategies have been applied to control weeds in agricultural and industrial areas. Knowledge regarding the effects of these methods on roadside vegetation is still poor. A 2-year field experiment was performed along a road located near Livorno (Tuscany, central Italy). Eight plots/strips were identified, of which four were subjected to periodical mechanical mowing and the remaining four were treated with a chemical herbicide based on glyphosate (the producer's recommended rates were used for the selective control of broad-leaved weeds). Our results clearly showed that roadside soil and vegetation are a significant reservoir of anthropogenic activities which have a strong negative effect on several phytosociological, pedochemical and biological parameters. Compared with conventional mechanical mowing, chemical treatment induced (i) a significant increase in organic matter in the upper plot layers (+18%), and (ii) a marked reduction in weed height throughout the entire period of the experiment. Irrespectively of the kind of treatment, no significance differences were detected in terms of (i) biological quality of soil (the abundance and diversity of arthropod communities did not change), and (ii) plant elemental content (bulk concentrations of analysed trace elements had a good fit within ranges of occurrence in the "reference plant"). The glyphosate partially controlled broad-leaved weeds and this moderate efficacy is dependent upon the season/time of application. In conclusion, the rational and sustainable use of chemical herbicides may be a useful tool for the management of roadside vegetation.

Can Ozone Alter the Terpenoid Composition and Membrane Integrity of in vitro Melissa officinalis Shoots?

Ozone affects volatile organic compounds that protect plants from biotic and abiotic stress. In vitro Melissa officinalis shoots were exposed to ozone (200 ppb, 3 h) in controlled environmental conditions: leaf pigments, membrane integrity and headspace composition were assayed during fumigation and after the recovery period (3 h from the beginning of the exposure, FBE). At the end of the exposure, no injury was observed in untreated and treated shoots, although an evident increase in lipid peroxidation was reported (+38.5 and +37.2% of TBARS levels in comparison with controls, respectively after 1 and 3 h FBE). The levels of total carotenoids significantly rose as a normal response mechanism to oxidative stress. SPME-GS-MS analysis showed that, as a consequence of the fumigation, the trends in non-terpenoid compounds increased after 1 and 3 h FBE. This suggests that the concentration and the duration of the treatment were enough to cause a breakdown of cells (as evidenced by increased TBARS levels) and involves an association between volatile products of the lipoxygenase pathway (LOX products) and membrane degradation.