Biochar as a soil amendment in the tree establishment phase: What are the consequences for tree physiology, soil quality and carbon sequestration?

Trees play a pivotal role in the urban environment alleviating the negative impacts of urbanization, and for this reason, local governments have promoted strongly tree planting policies. However, poor soil quality and neglect tree maintenance (e.g., irrigation and fertilization) can seriously mine the plant health status during the tree establishment phase. The use of biochar to provide long-lasting C to the soil and, at the same time, improving soil properties (e.g., improved water holding capacity), soil enzymes activities and NPK concentrations, is a promising research field. Therefore, with a two-step experiment, the study aimed to assay the physiological responses of a commonly used urban tree species (Tilia × europaea L.) to 1.5 % (w/w) biochar amendment (B), and secondly, to assess the ability of trees, grown in biochar amended soil, to tolerate a period of drought. Biochar amendment increased P and K availability in the soil, resulting in higher P and K concentrations in B than control leaves, according to the leaf stage. This induced B trees, higher values in both total biomass than controls (+22 %) in well-watered plants. Moreover, the higher water availability in soil amended with biochar helped B trees to tolerate water stress, with better leaf photosynthetic performances and a faster recovery than stressed controls after the re-watering. This study highlights the dual function of the biochar, improving CO2 sequestration and soil properties, and at the same time, enhancing plant physiological responses to environmental constraints. The use of biochar at the tree planting, especially in an urban environment, is a feasible and environmentally sustainable strategy to improve the success during the tree establishment phase.

Effects of water stress on spectral reflectance of bermudagrass.

In the south-central Italy, during summer rainfall does not supply a sufficient amount of water. Therefore, irrigation management during dry periods is important for maintaining turf quality. The hybrid bermudagrass (Cynodon dactylon (L.) Pers. × Cynodon transvaalensis Burtt-Davy) is known to represent the dominant warm-season turfgrass in warm to temperate climatic regions and its drought tolerance make bermudagrass a competitive turfgrass. A greenhouse experiment was conducted using uniform cores of hybrid bermudagrass, which were secured in a polyvinyl chloride cylinders and watered by constant sub-irrigation. The objectives of the present research were to measure the spectral reflectance with a new generation handheld spectroradiometer on hybrid bermudagrass and to explore various vegetation indices to be used as future detecting tool to study water stress in bermudagrass. Moreover, the potential uses of multivariate processing techniques for discriminating different water stress conditions in turfgrass has been investigated. Besides spectral indices, multivariate methods, although performed on a data set limited in terms of sample size, have shown a great potential for water stress monitoring in turfgrass and surely deserve further investigations. There are different indices that use distinct water absorption features independent of chlorophyll concentration, such as water index (WI = R900/R970) that has been reported to be a robust index of canopy water content and is used as an active indicator of changes in Leaf Relative Water Content (LRWC). Also, the ratio of WI with NDVI (WI/NDVI = (R900/R970)/((R800 - R680)/(R800 + R680)]) was found to be an effective indicator of water stress. Another vegetation index to detect water features is normalized difference water index (NDWI), designed to maximize reflectance of water by using green wavelengths. In our trial in bermudagrass the relationships studied, suggest that WI (900/970) and WI/NDVI, among the indices studied, are the more effective indicators of water stress. In fact, lower values of WI indicate higher water stress, while higher values of WI/NDVI indicate higher water stress levels.

Girled-induced anthocyanin accumulation in red-leafed Prunus cerasifera: Effect on photosynthesis, photoprotection and sugar metabolism.

The feedback regulation of photosynthesis depends on the cooperation of multiple signals, including sugars. Herein, the effect of shoot girdling was monitored on a daily basis for three days in green- and red-leafed Prunus cerasifera plants (GLP and RLP, respectively). The effect of anthocyanin presence was investigated in terms of photosynthesis, sugar metabolism and photoprotection. Net photosynthesis (A390) and stomatal conductance were reduced on the first day at 12:00 only in the girdled GLP (29 and 33 %, respectively). Moreover, the girdled GLP displayed at 12:00 higher sucrose, glucose and fructose concentrations than control leaves. Conversely, girdled RLP showed the first reduction of A390 at 18:00, with no significant differences at 12:00 in sucrose and glucose concentrations. The increased biosynthesis of anthocyanins that was only detected in girdled RLP contributed to lowering the accumulation of hexoses. Overall, these results revealed a sugar-buffering role exerted by anthocyanins that positively influence the feedback regulation of photosynthesis. Moreover, non-photochemical quenching, namely pNPQ, revealed the ability of anthocyanins to photoprotect photosystem II from supernumerary photons reaching the chloroplast, whose function was compromised by girdling. The present study provides a starting point to understand the possible link between photosynthesis regulation through sugar signalling and anthocyanin upregulation.

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.

Early Detection of Sage (Salvia officinalis L.) Responses to Ozone Using Reflectance Spectroscopy.

Advancements in techniques to rapidly and non-destructively detect the impact of tropospheric ozone (O3) on crops are required. This study demonstrates the capability of full-range (350-2500 nm) reflectance spectroscopy to characterize responses of asymptomatic sage leaves under an acute O3 exposure (200 ppb for 5 h). Using partial least squares regression, spectral models were developed for the estimation of several traits related to photosynthesis, the oxidative pressure induced by O3, and the antioxidant mechanisms adopted by plants to cope with the pollutant. Physiological traits were well predicted by spectroscopic models (average model goodness-of-fit for validation (R2): 0.65-0.90), whereas lower prediction performances were found for biochemical traits (R2: 0.42-0.71). Furthermore, even in the absence of visible symptoms, comparing the full-range spectral profiles, it was possible to distinguish with accuracy plants exposed to charcoal-filtered air from those exposed to O3. An O3 effect on sage spectra was detectable from 1 to 5 h from the beginning of the exposure, but ozonated plants quickly recovered after the fumigation. This O3-tolerance was confirmed by trends of vegetation indices and leaf traits derived from spectra, further highlighting the capability of reflectance spectroscopy to early detect the responses of crops to O3.

Discerning between Two Tuscany (Italy) Ancient Apple cultivars, 'Rotella' and 'Casciana', through Polyphenolic Fingerprint and Molecular Markers.

Ancient apple cultivars usually have higher nutraceutical value than commercial ones, but in most cases their variability in pomological traits does not allow us to discriminate among them. Fruit of two Tuscany ancient apple cultivars, 'Casciana' and 'Rotella', picked from eight different orchards (four for each cultivar) were analyzed for their pomological traits, organoleptic qualities, polyphenolic profile and antiradical activity. The effectiveness of a polyphenol-based cluster analysis was compared to molecular markers (internal transcribed spacers, ITS1 and ITS2) to unequivocally discern the two apples. 'Casciana' and 'Rotella' fruit had a higher nutraceutical value than some commercial cultivars, in terms of phenolic abundance, profile and total antiradical activity. Although pedo-climatic conditions of different orchards influenced the phenolic profile of both apples, the polyphenolic discriminant analysis clearly separated the two cultivars, principally due to higher amounts of procyanidin B2, procyanidin B3 and p-coumaroylquinic acid in 'Casciana' than in 'Rotella' fruit. These three polyphenols can be used proficiently as biochemical markers for distinguishing the two apples when pomological traits cannot. Conversely, ITS1 and ITS2 polymorphism did not allow us to distinguish 'Casciana' from 'Rotella' fruit. Overall, the use of polyphenolic fingerprint might represent a valid tool to ensure the traceability of products with a high economic value.

Ancient apple cultivars from Garfagnana (Tuscany, Italy): A potential source for 'nutrafruit' production.

Ancient apple cultivars are known for their organoleptic properties over a small geographic area, but little is known of their nutraceutical properties, which might be useful in large-scale breeding programmes. Nine ancient apple cultivars from Tuscany (Italy) were characterized for their organoleptic properties, phenolic profiles, and antioxidant activity. These cultivars had high polyphenol concentrations (principally flavanols and phenolic acids) and high total antioxidant capacity compared with most commercial apple cultivars. Fruits from the cultivars 'San Michele' and 'Del Debbio' showed a good compromise between fruit size and solid soluble content, and might be suitable for fresh consumption, while fruit from 'Benito', 'Della Piastra', 'Lugliese Grisanti', 'Del Sangue' and 'Ruggine' had high polyphenol contents and excellent antioxidant capacity, and may be suitable for breeding programmes. 'Ruggine' fruit could also be used for sweet juices with good nutraceutical properties due to their high soluble solid content and high flavanol concentration.

When "thirsty" means "less able to activate the signalling wave trigged by a pulse of ozone": A case of study in two Mediterranean deciduous oak species with different drought sensitivity.

There is a lack of knowledge about the possibility that plants facing abiotic stressors, such as drought, have an altered perception of a pulse of O3 and incur in alterations of their signalling network. This poses some concerns as to whether defensive strategy to cope episodic O3 peaks in healthy plants may fail under stress. In this study, a set of saplings of two Mediterranean deciduous species, Quercus cerris and Q. pubescens, was subjected to water withholding (20% of daily evapotranspiration for 15 days) while another set was kept well-watered. Saplings were then subjected to a pulse of O3 (200 nl l-1 for 5 h) or maintained in filtered air. Q. pubescens had a more severe decline of photosynthesis and leaf PDΨw (about -65% and 5-fold lower than in well-watered ones) and events of cell death were observed under drought when compared to Q. cerris, which is supportive for a higher sensitivity to drought exhibited by this species. When O3 was applied after drought, patterns of signalling compounds were altered in both species. Only in Q. pubescens, the typical O3-induced accumulation of apoplastic reactive oxygen species, which is the first necessary step for the activation of signalling cascade, was completely lost. In Q. cerris the most frequent changes encompassed the weakening of peaks of key signalling molecules (ethylene and salicylic acid), whereas in Q. pubescens both delayed (salicylic and jasmonic acid) or weakened (ethylene and salicylic acid) peaks were observed. This is translated to a higher ability of Q. cerris to maintain a prompt activation of defensive reaction to counteract oxidative damage due to the pollutant. Our results reveal the complexity of the signalling network in plants facing multiple stresses and highlight the need to further investigate possible alteration of defensive mechanism of tree species to predict their behavior.

How Quercus ilex L. saplings face combined salt and ozone stress: a transcriptome analysis.

BACKGROUND: Similar to other urban trees, holm oaks (Quercus ilex L.) provide a physiological, ecological and social service in the urban environment, since they remove atmospheric pollution. However, the urban environment has several abiotic factors that negatively influence plant life, which are further exacerbated due to climate change, especially in the Mediterranean area. Among these abiotic factors, increased uptake of Na + and Cl - usually occurs in trees in the urban ecosystem; moreover, an excess of the tropospheric ozone concentration in Mediterranean cities further affects plant growth and survival. Here, we produced and annotated a de novo leaf transcriptome of Q. ilex as well as transcripts over- or under-expressed after a single episode of O3 (80 nl l-1, 5 h), a salt treatment (150 mM for 15 days) or a combination of these treatments, mimicking a situation that plants commonly face, especially in urban environments. RESULTS: Salinity dramatically changed the profile of expressed transcripts, while the short O3 pulse had less effect on the transcript profile. However, the short O3 pulse had a very strong effect in inducing over- or under-expression of some genes in plants coping with soil salinity. Many differentially regulated genes were related to stress sensing and signalling, cell wall remodelling, ROS sensing and scavenging, photosynthesis and to sugar and lipid metabolism. Most differentially expressed transcripts revealed here are in accordance with a previous report on Q. ilex at the physiological and biochemical levels, even though the expression profiles were overall more striking than those found at the biochemical and physiological levels. CONCLUSIONS: We produced for the first time a reference transcriptome for Q. ilex, and performed gene expression analysis for this species when subjected to salt, ozone and a combination of the two. The comparison of gene expression between the combined salt + ozone treatment and salt or ozone alone showed that even though many differentially expressed genes overlap all treatments, combined stress triggered a unique response in terms of gene expression modification. The obtained results represent a useful tool for studies aiming to investigate the effects of environmental stresses in urban-adapted tree species.

Metabolic plasticity in the hygrophyte Moringa oleifera exposed to water stress.

Over the past decades, introduction of many fast-growing hygrophilic, and economically valuable plants into xeric environments has occurred. However, production and even survival of these species may be threatened by harsh climatic conditions unless an effective physiological and metabolic plasticity is available. Moringa oleifera Lam., a multipurpose tree originating from humid sub-tropical regions of India, is widely cultivated in many arid countries because of its multiple uses. We tested whether M. oleifera can adjust primary and secondary metabolism to efficiently cope with increasing water stress. It is shown that M. oleifera possesses an effective isohydric behavior. Water stress induced a quick and strong stomatal closure, driven by abscisic acid (ABA) accumulation, and leading to photosynthesis inhibition with consequent negative effects on biomass production. However, photochemistry was not impaired and maximal fluorescence and saturating photosynthesis remained unaffected in stressed leaves. We report for the first time that M. oleifera produces isoprene, and show that isoprene emission increased three-fold during stress progression. It is proposed that higher isoprene biosynthesis helps leaves cope with water stress through its antioxidant or membrane stabilizing action, and also indicates a general MEP (methylerythritol 4-phosphate) pathway activation that further helps protect photosynthesis under water stress. Increased concentrations of antioxidant flavonoids were also observed in water stressed leaves, and probably cooperate in limiting irreversible effects of the stress in M. oleifera leaves. The observed metabolic and phenotypic plasticity may facilitate the establishment of M. oleifera in xeric environments, sustaining the economic and environmental value of this plant.

Multiple Consequences Induced by Epidermally-Located Anthocyanins in Young, Mature and Senescent Leaves of Prunus.

Anthocyanic morphs are generally less efficient in terms of carbon gain, but, in turn, are more photoprotected than anthocyanin-less ones. To date, mature leaves of different morphs or leaves at different developmental stages within the same species have generally been compared, whereas there is a lack of knowledge regarding different stages of development of red vs. green leaves. Leaves (1-, 7-, and 13-week-old) of red- (RLP) and green-leafed (GLP) Prunus in terms of photosynthetic rate, carbon metabolism and photoprotective mechanisms were compared to test whether anthocyanin-equipped leaves perform better than anthocyanin-less leaves and whether photoprotection is the primary role of epidermally-located anthocyanins, using for the first time a recently-developed parameter of chlorophyll fluorescence (qPd). GLP leaves had a higher photosynthetic rate in 1- and 7-week-old leaves, but RLP leaves performed better at an early stage of senescence and had a longer leaf lifespan. Anthocyanins contributed to leaf photoprotection throughout the leaf development, but were tightly coordinated with carotenoids. Besides photoprotecting, we propose that epidermal anthocyanins may be principally synthetized to maintain an efficient carbon-sink strength in young and senescent leaves, thus extending the RLP leaf lifespan.

Overcoming limitation of "recalcitrant areas" to phytoextraction process: The synergistic effects of exogenous cytokinins and nitrogen treatments.

The aim of the present work was to test the efficiency of the phytoextraction process involving the use of exogenous phytohormone (cytokinins, CKs) and fertilizer (nitrogen, N) treatments in phytotechnologies to address risk management in "recalcitrant areas". The CKs and N treatments, alone or combined (CKs + N) in a Modulated Application (MA), were tested on the crop plant Helianthus annuus, common to Mediterranean area, fast growing and with high biomass production. Plants were grown on boron (B) contaminated sediments (collected from a geothermal area located in Tuscany (Italy). Plant growth, B uptake, together with plant stress parameters were investigated. Boron is easily taken up and translocated by some crop plants, but the high phytotoxicity can dramatically impact the plant growth and consequently the applicability and efficiency of the phytoextraction process. As indicators of plant stress, oxidative balance and photosynthetic parameters were investigated to give a deeper insight of phytotoxic mechanisms. Results showed that while each treatment (CKs and N alone) had significantly positive effects on plant health, the MA treatment provided a synergistic effect on morphological parameters and biomass production as a whole. After MA treatment, plants showed antioxidant activity comparable to that of the control (unpolluted sediments) and showed an increase of net photosynthesis. Moreover, our data showed very high values of B uptake and translocation (about 800 mg kg-1 in shoots), without any alteration triggered by the treatments (CKs and N alone or combined in MA). B phytoextraction resulted increased about fivefold with the MA treatments, while each treatment alone increased only two or three folds when treated with either CKs or N. The MA treatment is not "contaminant specific", so it could be applied in other "recalcitrant areas" where different types of contaminations occur, in order to overcome limitations of plant growth.

High innate attractiveness to black targets in the blue blowfly, Calliphora vomitoria (L.) (Diptera: Calliphoridae).

Calliphora vomitoria is a myiasis-causing fly in many animal species including humans. The control of blowflies is still anchored on the use of chemicals. However, mass trapping and lure-and-kill techniques represent a promising alternative to pesticides. Visual and olfactory cues are the main stimuli routing the fly's landing behavior. Notably, color attractiveness has been barely explored in flies of medical and veterinary importance, with special reference to blowflies. In this study, we investigated the innate color preferences in C. vomitoria adults, testing binary combinations of painted targets under laboratory conditions. The identity of tested species C. vomitoria was confirmed by DNA sequencing (18S and cox1 genes). C. vomitoria flies showed a significant preference for black colored targets in all tested binary color combinations, after 5, 15, 30 and 60 min of exposure. Black targets were significantly preferred over blue, red, yellow and white ones. Spectral characteristics of all tested color combinations were quantified and the innate attraction of blowflies towards black targets was discussed in relation to their behavioral ecology. To the best of our knowledge, this is the first report on innate color preferences in the Calliphora genus. Our findings can be useful to develop new, cheap and reliable monitoring traps as well as "lure and kill" tools to control blowfly pests.

Living in a Mediterranean city in 2050: broadleaf or evergreen 'citizens'?

The predicted effects of global change (GC) will be exacerbated in the more densely populated cities of the future, especially in the Mediterranean basin where some environmental cues, such as drought and tropospheric ozone (O3) pollution, already mine seriously plant survival. Physiological and biochemical responses of a Mediterranean, evergreen, isohydric plant species (Quercus ilex) were compared to those of a sympatric, deciduous, anisohydric species (Q. pubescens) under severe drought (20% of the effective daily evapotranspiration) and/or chronic O3 exposure (80 ppb for 5 h day-1 for 28 consecutive days) to test which one was more successful in those highly limiting conditions. Results show that (i) the lower reduction of total leaf biomass of Q. ilex as compared to Q. pubescens when subjected to drought and drought × O3 (on average -59 vs -70%, respectively); (ii) the steeper decline of photosynthesis found in Q. pubescens under drought (-87 vs -81%) and drought × O3 (-69 vs -59%, respectively); (iii) the increments of malondialdehyde (MDA) by-products found only in drought-stressed Q. pubescens; (iv) the impact of O3, found only in Q. pubescens leaves and MDA, can be considered the best probes of the superiority of Q. ilex to counteract the effect of mild-severe drought and O3 stress. Also, an antagonistic effect was found when drought and O3 were applied simultaneously, as usually happens during typical Mediterranean summers. Our dataset suggests that on future, the urban greening should be wisely pondered on the ability of trees to cope the most impacting factors of GC, and in particular their simultaneity.

Losing the Warning Signal: Drought Compromises the Cross-Talk of Signaling Molecules in Quercus ilex Exposed to Ozone.

Understanding the interactions between drought and acute ozone (O3) stress in terms of signaling molecules and cell death would improve the predictions of plant responses to climate change. The aim was to investigate whether drought stress influences the responses of plants to acute episodes of O3 exposure. In this study, the behavior of 84 Mediterranean evergreen Quercus ilex plants was evaluated in terms of cross-talk responses among signaling molecules. Half of the sample was subjected to drought (20% of the effective daily evapotranspiration, for 15 days) and was later exposed to an acute O3 exposure (200 nL L-1 for 5 h). First, our results indicate that in well-water conditions, O3 induced a signaling pathway specific to O3-sensitive behavior. Second, different trends and consequently different roles of phytohormones and signaling molecules (ethylene, ET; abscisic acid, ABA; salycilic acid, SA and jasmonic acid, JA) were observed in relation to water stress and O3. A spatial and functional correlation between these signaling molecules was observed in modulating O3-induced responses in well-watered plants. In contrast, in drought-stressed plants, these compounds were not involved either in O3-induced signaling mechanisms or in leaf senescence (a response observed in water-stressed plants before the O3-exposure). Third, these differences were ascribable to the fact that in drought conditions, most defense processes induced by O3 were compromised and/or altered. Our results highlight how Q. ilex plants suffering from water deprivation respond differently to an acute O3 episode compared to well-watered plants, and suggest new effect to be considered in plant responses to environmental changes. This poses the serious question as to whether or not multiple high-magnitude O3 events (as predicted) can change these cross-talk responses, thus opening it up possible further investigations.

Multiple cues produced by a robotic fish modulate aggressive behaviour in Siamese fighting fishes.

The use of robotics to establish social interactions between animals and robots, represents an elegant and innovative method to investigate animal behaviour. However, robots are still underused to investigate high complex and flexible behaviours, such as aggression. Here, Betta splendens was tested as model system to shed light on the effect of a robotic fish eliciting aggression. We evaluated how multiple signal systems, including a light stimulus, affect aggressive responses in B. splendens. Furthermore, we conducted experiments to estimate if aggressive responses were triggered by the biomimetic shape of fish replica, or whether any intruder object was effective as well. Male fishes showed longer and higher aggressive displays as puzzled stimuli from the fish replica increased. When the fish replica emitted its full sequence of cues, the intensity of aggression exceeded even that produced by real fish opponents. Fish replica shape was necessary for conspecific opponent perception, evoking significant aggressive responses. Overall, this study highlights that the efficacy of an artificial opponent eliciting aggressive behaviour in fish can be boosted by exposure to multiple signals. Optimizing the cue combination delivered by the robotic fish replica may be helpful to predict escalating levels of aggression.

The harsh life of an urban tree: the effect of a single pulse of ozone in salt-stressed Quercus ilex saplings.

Ozone (O3) and salinity are usually tested as combined factors on plant performance. However, the response to a single episode of O3 in plants already stressed by an excess of NaCl as occurs in the natural environment has never been investigated, but is important given that it is commonly experienced in Mediterranean areas. Three-year-old Quercus ilex L. (holm oak) saplings were exposed to salinity (150 mM NaCl, 15 days), and the effect on photosynthesis, hydric relations and ion partitioning was evaluated (Experiment I). In Experiment II, salt-treated saplings were exposed to 80 nl l-1 of O3 for 5 h, which is a realistic dose in a Mediterranean environment. Gas exchanges, chlorophyll fluorescence and antioxidant systems were characterized to test whether the salt-induced stomatal closure limited O3 uptake and stress or whether the pollutant represents an additional stressor for plants. Salt-dependent stomatal closure depressed the photosynthetic process (-71.6% of light-saturated rate of photosynthesis (A380)) and strongly enhanced the dissipation of energy via the xanthophyll cycle. However, salt-treated plants had higher values of net assimilation rate/stomatal conductance (A/gs) than the controls, which was attributable to a greater mesophyll conductance gm/gs and carboxylation efficiency (higher gm/maximal rate of Rubisco carboxylation (Vcmax)), thus suggesting no damage to chloroplasts. O3 did not exacerbate the effect of salinity on photosynthesis, however a general enhancement of the Halliwell-Asada cycle was necessary to counteract the O3-triggered oxidative stress. Despite the 79.4% gs reduction in salt-stressed plants, which strongly limited the O3 uptake, a single peak in the air pollutant led to an additional burden for the antioxidant system when plants had been previously subjected to salinity.

Salt-tolerant rootstock increases yield of pepper under salinity through maintenance of photosynthetic performance and sinks strength.

The performance of a salt-tolerant pepper (Capsicum annuum L.) accession (A25) utilized as a rootstock was assessed in two experiments. In a first field experiment under natural salinity conditions, we observed a larger amount of marketable fruit (+75%) and lower Blossom-end Root incidence (-31%) in commercial pepper cultivar Adige (A) grafted onto A25 (A/A25) when compared with ungrafted plants. In order to understand this behavior a second greenhouse experiment was conducted to determine growth, mineral partitioning, gas exchange and chlorophyll a fluorescence parameters, antioxidant systems and proline content in A and A/A25 plants under salinity conditions (80 mM NaCl for 14 days). Salt stress induced significantly stunted growth of A plants (-40.6% of leaf dry weight) compared to the control conditions, while no alterations were observed in A/A25 at the end of the experiment. Accumulation of Na(+) and Cl(-) in leaves and roots was similar in either grafted or ungrafted plants. Despite the activation of protective mechanisms (increment of superoxide dismutase, catalase, ascorbate peroxidase activity and non-photochemical quenching), A plants showed severely reduced photosynthetic CO2 assimilation (-45.6% of AN390) and substantial buildup of malondialdehyde (MDA) by-product, suggesting the inability to counteract salt-triggered damage. In contrast, A/A25 plants, which had a constitutive enhanced root apparatus, were able to maintain the shoot and root growth under salinity conditions by supporting the maintained photosynthetic performance. No increases in catalase and ascorbate peroxidase activities were observed in response to salinity, and MDA levels increased only slightly; indicating that alleviation of oxidative stress did not occur in A/A25 plants. In these plants the increased proline levels could protect enzymatic stability from salt-triggered damage, preserving the photosynthetic performance. The results could indicate that salt stress was vanished by the lack of negative effects on photosynthesis that support the maintained plant growth and increased marketable yield of the grafted plants.

Variations in physiological and biochemical traits of oak seedlings grown under drought and ozone stress.

Despite the huge biodiversity characterizing the Mediterranean environment, environmental constraints, such as high sunlight and high temperatures alongside with dry periods, make plant survival hard. In addition, high irradiance leads to increasing ozone (O3 ) concentrations in ambient air. In this era of global warming, it is necessary to understand the mechanisms that allow native species to tolerate these environmental constraints and how such mechanisms interact. Three Mediterranean oak species (Quercus ilex, Quercus pubescens and Quercus cerris) with different features (drought tolerant, evergreen or deciduous species) were selected to assess their biometrical, physiological and biochemical responses under drought and/or O3 stress (80-100 nl l(-1) of O3 for 5 h day(-1) for 77 consecutive days). Leaf visible injury appeared only under drought stress (alone or combined with O3 ) in all three species. Drought × O3 induced strong reductions in leaf dry weight in Q. pubescens and Q. cerris (-70 and -75%, respectively). Alterations in physiological (i.e. decrease in maximum carboxylation rate) and biochemical parameters (i.e. increase in proline content and build-up of malondialdehyde by-products) occurred in all the three species, although drought represented the major determinant. Quercus ilex and Q. pubescens, which co-occur in dry environments, were more tolerant to drought and drought × O3 . Quercus ilex was the species in which oxidative stress occurred only when drought was applied with O3 . High plasticity at a biochemical level (i.e. proline content) and evergreen habitus are likely on the basis of the higher tolerance of Q. ilex.

Epidermal coumaroyl anthocyanins protect sweet basil against excess light stress: multiple consequences of light attenuation.

The putative photoprotective role of foliar anthocyanins continues to attract heated debate. Strikingly different experimental set-ups coupled with a poor knowledge of anthocyanin identity have likely contributed to such disparate opinions. Here, the photosynthetic responses to 30 or 100% solar irradiance were compared in two cultivars of basil, the green-leafed Tigullio (TG) and the purple-leafed Red Rubin (RR). Coumaroyl anthocyanins in RR leaf epidermis significantly mitigated the effects of high light stress. In full sunlight, RR leaves displayed several shade-plant traits; they transferred less energy than did TG to photosystem II (PSII), and non-photochemical quenching was lower. The higher xanthophyll cycle activity in TG was insufficient to prevent inactivation of PSII in full sunlight. However, TG was the more efficient in the shade; RR was far less able to accommodate a large change in irradiance. Investment of carbon to phenylpropanoid biosynthesis was more in RR than in TG in the shade, and was either greatly enhanced in TG or varied little in RR because of high sunlight. The metabolic cost of photoprotection was lower whereas light-induced increase in biomass production was higher in RR than in TG, thus making purple basil the more light tolerant. Purple basil appears indeed to display the conservative resource-use strategy usually observed in highly stress tolerant species. We conclude that the presence of epidermal coumaroyl anthocyanins confers protective benefits under high light, but it is associated with a reduced plasticity to accommodate changing light fluxes as compared with green leaves.