Sewage sludge amendment of rice as a potential alternative to mineral fertilizer: Analyses of physiological, biochemical and molecular mechanisms of plant response.

Sewage sludge (SS) disposal poses environmental concerns, yet its organic matter, macro- and micronutrients, make it potentially beneficial for enhancing soil quality and crop yield. This study focuses on three types of SS: "R10" (SS1), which is commonly used in agricultural practices, and two environmentally friendlier options (SS2 and SS3), as alternatives to mineral fertilizer (urea) for rice cultivation. A pot experiment was conducted to investigate the ecophysiological, biochemical, and molecular responses of rice at three different growth stages. SS application led to a significant increase in biomass production (particularly SS3), along with increased nitrogen (N) levels. Enhanced chlorophyll content was observed in SS-treated plants, especially during inflorescence emergence (with the highest content in SS3 plants). At the ecophysiological and biochemical levels, SS treatments did not adversely affect plant health, as evidenced by unchanged values of maximal PSII photochemical efficiency and malondialdehyde by-products. At biochemical and gene expression levels, antioxidant enzyme activities showed transient variations, likely related to physiological adjustments rather than oxidative stress. Ascorbic acid and glutathione did not significantly vary. This study concludes that the use of SS in soil can be a viable alternative fertilizer for rice plants, with positive effects on biomass, chlorophyll content, and no adverse effects on plant health. Among the tested SSs, SS3 showed the most positive effect, even compared to commercial fertilizer. These results suggest that SS application could improve rice yield while addressing environmental concerns surrounding SS disposal.

Phytotoxic Ozone Dose-Response Relationships for Durum Wheat (Triticum durum, Desf.).

Ozone (O3) pollution poses a significant threat to global crop productivity, particularly for wheat, one of the most important staple foods. While bread wheat (Triticum aestivum) is unequivocally considered highly sensitive to O3, durum wheat (Triticum durum) was often found to be more tolerant. This study investigated the O3 dose-response relationships for durum wheat in the Mediterranean region, focusing mainly on grain yield losses, and utilizing the phytotoxic ozone dose (POD) metric to describe the intensity of the stressor. The results from two experiments with Open-Top Chambers performed in 2013 and 2014 on two relatively sensitive durum wheat cultivars confirmed that this wheat species is far more tolerant than bread wheat. The use of a local parameterization of a stomatal conductance model based on field measurements did not significantly improve the dose-response relationships obtained in comparison to the generic parameterization suggested by the Mapping Manual of the United Nations Economic Commission for Europe (UNECE). The POD6 critical level of 5 mmolO3 m-2 for 5% grain yield loss was remarkably higher than the one established for bread wheat with analogous experiments, highlighting that O3 risk assessments based on bread wheat may largely overestimate the damage in the Mediterranean region where durum wheat cultivation prevails.

Evaluation of Hybrid Models to Estimate Chlorophyll and Nitrogen Content of Maize Crops in the Framework of the Future CHIME Mission.

In the next few years, the new Copernicus Hyperspectral Imaging Mission (CHIME) is foreseen to be launched by the European Space Agency (ESA). This missions will provide an unprecedented amount of hyperspectral data, enabling new research possibilities within several fields of natural resources, including the "agriculture and food security" domain. In order to efficiently exploit this upcoming hyperspectral data stream, new processing methods and techniques need to be studied and implemented. In this work, the hybrid approach (HYB) and its variant, featuring sampling dimensionality reduction through active learning heuristics (HAL), were applied to CHIME-like data to evaluate the retrieval of crop traits, such as chlorophyll and nitrogen content at both leaf (LCC and LNC) and canopy level (CCC and CNC). The results showed that HYB was able to provide reliable estimations at canopy level (R2 = 0.79, RMSE = 0.38 g m-2 for CCC and R2 = 0.84, RMSE = 1.10 g m-2 for CNC) but failed at leaf level. The HAL approach improved retrieval accuracy at canopy level (best metric: R2 = 0.88 and RMSE = 0.21 g m-2 for CCC; R2 = 0.93 and RMSE = 0.71 g m-2 for CNC), providing good results also at leaf level (best metrics: R2 = 0.72 and RMSE = 3.31 µg cm-2 for LCC; R2 = 0.56 and RMSE = 0.02 mg cm-2 for LNC). The promising results obtained through the hybrid approach support the feasibility of an operational retrieval of chlorophyll and nitrogen content, e.g., in the framework of the future CHIME mission. However, further efforts are required to investigate the approach across different years, sites and crop types in order to improve its transferability to other contexts.

Oxidative Stress Mitigation by Chitosan Nanoparticles in Durum Wheat Also Affects Phytochemicals and Technological Quality of Bran and Semolina.

In our previous work, durum wheat cv. Fabulis was grown over two consecutive seasons (2016-2017 and 2017-2018) in an experimental field in the north of Italy. With the aim of mitigating oxidative stress, plants were subjected to four treatments (deionized water, CHT 0.05 mg/mL, CHT-NPs, and CHT-NPs-NAC) three times during the experiment. Chitosan nanoparticles (CHT-NPs) reduced symptom severity on wheat leaves and positively influenced the final grain yield. The present work aimed at investigating whether CHT treatments and particularly N-acetyl cysteine (NAC)-loaded or -unloaded CHT-NPs, while triggering plant defense mechanisms, might also vary the nutritional and technological quality of grains. For this purpose, the grains harvested from the previous experiment were analyzed for their content in phytochemicals and for their technological properties. The results showed that CHT increased the polyphenol and tocopherol content and the reducing capacity of bran and semolina, even if the positive effect of the nano-formulation remained still unclear and slightly varied between the two years of cultivation. The positive effect against oxidative stress induced by the chitosan treatments was more evident in the preservation of both the starch pasting properties and gluten aggregation capacity, indicating that the overall technological quality of semolina was maintained. Our data confirm the role of chitosan as an elicitor of the antioxidant defense system in wheat also at the grain level.

Influence of year, genotype and cultivation system on nutritional values and bioactive compounds in tomato (Solanum lycopersicum L.).

Two tomato genotypes were grown in open field by three cultivation systems (one conventional and two distinct organic for mulching) in three years, 2015, 2016 and 2017. Yields, sugars, organic acids, amino acids, ascorbic acid, biothiols, carotenoids and phenols were measured. Weather conditions largely differed among harvest years, with summer 2016 rainier and less warm, and an opposite summer 2017. Organic systems had lower yields than conventional one but also, interestingly, lower waste percentages. Furthermore, tilled and no-tilled organic systems provided comparable yields. With respect to 3-year average, sugars were higher in 2017, acids in 2016 and in organic fruits, and amino acids increased in 2015 and in conventional samples. A higher glutathione content was found in organic samples, and higher carotenoids in 2017. Phenols increased in 2016, with a higher chlorogenic acid content in organic tomatoes. Some differences between genotypes were observed, highlighting their different adaptability to growing systems.

Chitosan Nanoparticles Loaded with N-Acetyl Cysteine to Mitigate Ozone and Other Possible Oxidative Stresses in Durum Wheat.

Modern durum wheat cultivars are more prone to ozone stress because of their high photosynthetic efficiency and leaf gas exchanges that cause a greater pollutant uptake. This, in turn, generates an increased reactive oxygen species (ROS) production that is a challenge to control by the antioxidant system of the plant, therefore affecting final yield, with a reduction up to 25%. With the aim of mitigating oxidative stress in wheat, we used chitosan nanoparticles (CHT-NPs) either unloaded or loaded with the antioxidant compound N-acetyl cysteine (NAC), on plants grown either in a greenhouse or in an open field. NAC-loaded NPs were prepared by adding 0.5 mg/mL NAC to the CHT solution before ionotropic gelation with tripolyphosphate (TTP). Greenhouse experiments evidenced that CHT-NPs and CHT-NPs-NAC were able to increase the level of the leaf antioxidant pool, particularly ascorbic acid (AsA) content. However, the results of field trials, while confirming the increase in the AsA level, at least in the first phenological stages, were less conclusive. The presence of NAC did not appear to significantly affect the leaf antioxidant pool, although the grain yield was slightly higher in NAC-treated parcels. Furthermore, both NAC-loaded and -unloaded CHT-NPs partially reduced the symptom severity and increased the weight of 1000 seeds, thus showing a moderate mitigation of ozone injury.

Hop Leaves as an Alternative Source of Health-Active Compounds: Effect of Genotype and Drying Conditions.

In hop cultivation, one-third of the crop is a valuable product (hop cones), and two-thirds is unexploited biomass, consisting mainly of leaves and stems, which, in a circular economy approach, can be recovered and, once stabilized, supplied to industrial sectors, such as cosmetics, pharmaceuticals and phytotherapy, with high added value. In this regard, this study aimed to investigate the effects of two different drying methods: oven drying (OD) at 45 °C and freeze-drying (FD), on the overall nutraceutical profile (i.e., total phenols, total flavans and total thiols), pigment content (i.e., carotenoids and chlorophylls) and the antioxidant potential of leaves from five different Humulus lupulus varieties grown in central Italy. Moreover, attenuated total reflectance infrared (ATR-FTIR) spectroscopy was applied to dried leaf powders to study the influence of both the variety and treatment on their molecular fingerprints. The spectral data were then analyzed by principal component analysis (PCA), which was able to group the samples mainly based on the applied treatment. Considering the overall phytochemical profile, FD appeared to be the most suitable drying method, while OD provided higher carotenoid retention, depending on the genotype considered. Finally, unsupervised chemometric tools (i.e., PCA and hierarchical clustering) revealed that the two main clusters contained subclusters based on the drying treatment applied; these subgroups were related to the susceptibility of the variety to the drying conditions studied.

Phytochemical Characterization and In Vitro Antioxidant Properties of Four Brassica Wild Species from Italy.

In the present study, we evaluated for the first time the variability of antioxidant traits of four Brassica wild species: B. incana, B. macrocarpa, B. villosa, and B. rupestris. The content of the main water-soluble antioxidants (phenolics, ascorbic acid, and total biothiols) and the in vitro antioxidant potential (1,1-diphenyl-2-picrylhydrazil (DPPH) and superoxide anion scavenging capacity) were investigated. A total of 28 polyphenolic compounds were identified by LC/MS and quantitated by HPLC/DAD analysis. Kaempferol and quercetin derivatives were the most abundant phenolics compared to hydroxycinnamoyl gentiobiosides. In the ten populations, phenolics ranged from 163.9 to 533.9 mg/100 g dry weight (d.w.), ascorbic acid from 7.6 to 375.8 mg/100 g d.w., and total biothiols from 0.59 to 5.13 mg/100 g d.w. The different classes of phytochemicals were separated using solid-phase extraction at increasing methanol concentrations, and the antioxidant power of fractionated extracts was evaluated. The superoxide anion scavenging activity was significantly correlated to phenolics, particularly to flavonol derivatives, while DPPH was mainly related to ascorbic acid content. The present findings improve the knowledge of the phytochemical composition of Italian Brassica wild species by showing the great diversity of phytochemicals among populations and highlighting their importance as a valuable genetic resource for developing new cultivars with improved bioactive content.

Phenolic composition and antioxidant capacity of hawthorn (Crataegus oxyacantha L.) flowers and fruits grown in Algeria.

Background To the best of our knowledge, up to now, there are no reports on the antioxidant activity of phenolic compounds of Crataegus oxyacantha flowers and fruits. Such detailed information is essential to advance the existing knowledge and to promote the use of this species growing in Africa. Therefore, the objective of this investigation was to analyze the content of phenolic compounds as well as the antioxidant activities of flowers and fruits of C. oxyacantha from Bejaia (Northeastern Algeria) by different analytical methods, and to determine the relationship between them. Methods Phenolic compounds and antioxidant activity of Algerian hawthorn flowers and fruits (C. oxyacantha L.) were studied. Total phenolics, flavonoids and flavonols were determined using colorimetric methods. The phenolic profile was analyzed by high-performance liquid chromatography with diode array detection, and the in vitro antioxidant activity was measured using scavenging assay (FRAP) and Fremy's salt, using electronic paramagnetic resonance. Results Different classes of phenolic compounds were identified and quantified. Hawthorn flowers contained higher levels of hydroxycinnamic acids, vitexin derivatives and flavonols compared to fruits, while anthocyanins were present only in fruits. Significant correlations were found between phenolic content and antioxidant activity. The results also showed that although the phenolic content of the two parts was different, their antioxidant capacity was not statistically different. Conclusions The results of this study indicate that hawthorn flowers and fruits of C. oxyacantha may be considered as a natural source of bioactive compounds.

Impact of year of harvest, genotype and cultivation method on bioactives and Pru d 1 allergen content in plums.

The present work studied the effect of the year of harvest, the genotype and the cultivation method on the nutritional quality and the allergen content of three plum cultivars. The common quality parameters and the phytochemical content strongly varied with the year and the cultivar, while the system of cultivation had a minor influence. In particular, ascorbic acid greatly decreased in 2016 compared to 2015, while polyphenols were higher in 2016. The health-promoting compounds, and particularly phenolics, were significantly correlated with the antioxidant capacity. Finally, the allergen content was strongly dependent on the content of flavan-3-ols, suggesting that this class of phenolics is determinant in influencing the allergen content in plums. Results showed that the major factor affecting the quality and the concentration of natural metabolites of plum, in addition to the diversity among genotypes, is the year-to-year variation, whereas the system of cultivation plays a marginal role.

The ozone-like syndrome in durum wheat (Triticum durum Desf.): Mechanisms underlying the different symptomatic responses of two sensitive cultivars.

Colombo and Sculptur are two modern durum wheat cultivars that, in previous studies, proved to be very sensitive to ozone injury in terms of eco-physiological parameters and significant grain yield loss. Nevertheless, their response regarding leaf visible symptoms was very different; Sculptur showed almost no symptoms, even after several weeks of ozone exposure, whereas Colombo showed in a few weeks typical ozone-like symptoms (chlorotic/necrotic spots). The mechanisms underlying this different response has been studied with a biochemical and microscopical approach. Plants were grown in Open-Top Chambers (OTCs) and exposed to charcoal filtered and ozone enriched air. Flag leaves were analyzed at two phenological stages (pre- and post-anthesis). At pre-anthesis the ascorbate pool was significantly lower in Colombo, which also underwent an increase in the oxidized glutathione content and abundant H2O2 deposition in mesophyll cells around the substomatal chamber. No or scarce H2O2 was found at both phenological stages in ozone exposed leaf tissues of Sculptur, where stomata appeared often closed. In this cultivar, transmission electron microscopy showed that chloroplasts in apparently undamaged mesophyll cells were slightly swollen and presented numerous plastoglobuli, as a result of a mild oxidative stress. These results suggest that Sculptur leaves remains symptomless as a consequence of the higher content of constitutive ascorbate pool and the synergistic effect of stomata closure. Instead, Colombo shows chlorotic/necrotic symptoms because of the lower ROS (Reactive Oxygen Species) scavenging capacity and the less efficient stomata closure that lead to severe damages of groups of the mesophyll cells, however leaving the surrounding photosynthetic tissue functional.

The effect of the germination temperature on the phytochemical content of broccoli and rocket sprouts.

This study investigates the effect of different germination temperatures (10, 20 and 30 °C) on the phytochemical content as well as reducing and antioxidant capacity of broccoli and rocket sprouts. In both seeds and sprouts, the total glucosinolates and ascorbic acid contents did not differ between vegetables, while broccoli exhibited exceptionally higher polyphenols and greater reducing and antioxidant capacity compared to rocket. In both species, an increase in germination temperature positively affected the glucosinolate content. Ascorbic acid increased during germination without a difference among the three tested temperatures. The phenol content in broccoli sprouts increased when they were grown at 30 °C, but the amount decreased at the highest temperatures in rocket. The reducing and antioxidant capacities increased with germination, and higher indexes were detected at 10 °C, particularly in rocket. Different germination temperatures differentiate the health-promoting phytochemical content and antioxidant properties in broccoli and rocket sprouts.

Variations in the phytochemical contents and antioxidant capacity of organically and conventionally grown Italian cauliflower (Brassica oleracea L. subsp. botrytis): results from a three-year field study.

A three-year field study (2009-2011) was performed to evaluate phytochemicals and antioxidant capacities of two genotypes (HF1 Emeraude and the local variety, Velox) of green cauliflower grown under organic and conventional management. The conventional system increased yield, but had little effect on the dry matter, whereas the organic system increased the soluble solids. Phytochemicals and antioxidant capacity showed significant year-to-year variability. During the third year, the scarce rainfall determined a significant increase of total glucosinolates and a general decrease of antioxidants in all samples. Interestingly, in the same year organic plants were less affected by the unfavorable climatic conditions, as they increased ascorbic acid, polyphenols, and carotenoids with respect to conventional ones. The overall results for the three years showed that the two genotypes responded differently. Compared to the conventional system, Velox showed 24, 21, 13, 48, and 44% higher content of ascorbic acid, polyphenols, carotenoids, volatiles, and antioxidant capacity, respectively. In contrast, no significant increase in the phytochemicals or the antioxidant potential was found in organic Emeraude, with the exception of total volatiles (+41%). These findings suggest that organic cultivation may be highly effective for particular cauliflower genotypes.

Analysis of transcript and metabolite levels in Italian rice (Oryza sativa L.) cultivars subjected to osmotic stress or benzothiadiazole treatment.

One of the major objectives of rice (Oryza sativa L.) breeding programs is the development of new varieties with higher tolerance/resistance to both abiotic and biotic stresses. In this study, Italian rice cultivars were subjected to osmotic stress or benzothiadiazole (BTH) treatments. An analysis of the expression of selected genes known to be involved in the stress response and (1)H nuclear magnetic resonance ((1)H NMR) metabolic profiling were combined with multivariate statistical analyses to elucidate potential correlations between gene expression or metabolite content and observed tolerant/resistant phenotypes. We observed that the expression of three chosen genes (two WRKY genes and one peroxidase encoding gene) differed between susceptible and resistant cultivars in response to BTH treatments. Moreover, the analysis of metabolite content, in particular in the osmotic stress experiment, enabled discrimination between selected cultivars based on differences in the accumulation of some primary metabolites, primarily sugars. This research highlights the potential usefulness of this approach to characterise rice varieties based on transcriptional or metabolic changes due to adverse environmental conditions.

Using optical remote sensing techniques to track the development of ozone-induced stress.

In this paper, a literature review about optical remote sensing (RS) of O(3) stress is presented. Studies on O(3)-induced effects on vegetation reflectance have been conducted since late '70s based on the analysis of optical RS data. Literature review reveals that traditional RS techniques were able to detect changes in leaf and canopy reflectance related to O(3)-induced stress when visible symptoms already occurred. Only recently, advanced RS techniques using hyperspectral sensors, demonstrated the feasibility of detecting the stress in its early phase by monitoring excess energy dissipation pathways such as chlorophyll fluorescence and non-photochemical quenching (NPQ). Steady-state fluorescence (Fs), measured by exploiting the Fraunhofer line depth principle and NPQ related xanthophyll-cycle, estimated through the photochemical reflectance index (PRI) responded to O(3) fumigation before visible symptoms occurred. This opens up new possibilities for the early detection of vegetation O(3) stress by means of hyperspectral RS.

Acute exposure of the aquatic macrophyte Callitriche obtusangula to the herbicide oxadiazon: the protective role of N-acetylcysteine.

In this study we investigated the acute exposure of the aquatic macrophyte Callitriche obtusangula to the herbicide oxadiazon (Ronstar). The toxic effects on C. obtusangula were evaluated, 24h after exposure, by assessing visible necrotic leaf lesions and, 12 h after exposure, via analyses of dead cells and hydrogen peroxide (H2O2) deposits localized by histocytochemical analysis with Trypan blue and 3,3'-diaminobenzidine (DAB), respectively. As a result, we found that 0.1275 microg L(-1) a.i. (active ingredient) oxadiazon was the maximum concentration that produced no observable adverse effects (NOAEC) both at leaf and tissue levels, at any considered exposure time. Additionally, we assayed the protective effect of pre-treatment with 0.25 mM N-acetylcysteine (NAC), a cysteine donor, on the damage caused by the toxic herbicidal dose of 6.37 microg L(-1) a.i to C. obtusangula, correlating the NAC observed protection to the direct H2O2-scavenging and to the enhancement of glutathione parameters. NAC-treated plants showed a fourfold increase in the GSH (reduced glutathione)+GSSG (oxidised glutathione) content (149.2 nmol g(-1) FW) compared to controls (36.1 nmol g(-1) FW); in the NAC+oxadiazon treatments, the GSH+GSSG content was more than fivefold higher (202.1 nmol g(-1) FW). GSH showed a similar trend in NAC and NAC+oxadiazon treatments, being six- (130.0 nmol g(-1) FW) and eightfold (185.0 nmol g(-1) FW) higher, respectively, compared to controls (20.7 nmol g(-1) FW). Accordingly, the GSH/GSSG ratio in NAC- and NAC+oxadiazon-treated plants was significantly increased compared to controls, indicating alleviation of oxidative stress.

Nutritional traits of bean ( Phaseolus vulgaris ) seeds from plants chronically exposed to ozone pollution.

The effect of chronic exposure to ozone pollution on nutritional traits of bean ( Phaseolus vulgaris L. cv. Borlotto Nano Lingua di Fuoco) seeds from plants grown in filtered and nonfiltered open-top chambers (OTCs) has been investigated. Results showed that, among seed macronutrients, ozone significantly raised total lipids, crude proteins, and dietary fiber and slightly decreased total free amino acid content, although with a significant reduction of asparagine, lysine, valine, methionine, and glycine, compensated by a conspicuous augmentation of ornithine and tryptophan. Phytosterol analysis showed a marked increase of beta-sitosterol, stigmasterol, and campesterol in seeds collected from nonfiltered OTCs. With regard to secondary metabolites, ozone exposure induced a slight increase of total polyphenol content, although causing a significant reduction of some flavonols (aglycone kaempferol and its 3-glucoside derivative) and hydroxycinnamates (caffeic, p-coumaric, and sinapic acids). Total anthocyanins decreased significantly, too. Nevertheless, ozone-exposed seeds showed higher antioxidant activity, with higher Trolox equivalent antioxidant capacity (TEAC) values than those measured in seeds collected from filtered air.

Assessing Steady-state Fluorescence and PRI from Hyperspectral Proximal Sensing as Early Indicators of Plant Stress: The Case of Ozone Exposure.

High spectral resolution spectrometers were used to detect optical signals ofongoing plant stress in potted white clover canopies subjected to ozone fumigation. Thecase of ozone stress is used in this manuscript as a paradigm of oxidative stress. Steadystatefluorescence (Fs) and the Photochemical Reflectance Index (PRI) were investigatedas advanced hyperspectral remote sensing techniques able to sense variations in the excessenergy dissipation pathways occurring when photosynthesis declines in plants exposed to astress agent. Fs and PRI were monitored in control and ozone fumigated canopies during a21-day experiment together with the traditional Normalized Difference Vegetation Index(NDVI) and physiological measurements commonly employed by physiologists to describestress development (i.e. net CO2 assimilation, active fluorimetry, chlorophyll concentrationand visible injuries). It is shown that remote detection of an ongoing stress through Fs andPRI can be achieved in an early phase, characterized by the decline of photosynthesis. Onthe contrary, NDVI was able to detect the stress only when damage occurred. These resultsopen up new possibilities for assessment of plant stress by means of hyperspectral remotesensing.