Olive-mill wastewater (OMW), an effluent of olive oil extraction process, is annually produced in huge amounts in olive growing areas. An interesting option for its disposal is the spreading on agricultural land, provided that phytotoxic effects are neutralized. The objective of the present investigation was to evaluate the potential of an enzyme-based treatment in removing OMW phytotoxicity. To this aim, germinability experiments on durum wheat (Triticum durum Desf. cv. Duilio) were conducted in the presence of different dilutions of raw or enzyme-treated OMW. OMW treatment with laccase resulted in a 65% and 86% reduction in total phenols and ortho-diphenols respectively, due their polymerization as revealed by size-exclusion chromatography. Raw OMW exerted a significant concentration-dependent inhibition on the germinability of durum wheat seeds which was evident up to a dilution rate of 1:8. When the effluent was treated with a fungal laccase, germinability was increased by 57% at a 1:8 dilution and by 94% at a 1:2 dilution, as compared to the same dilutions using untreated OMW. The treatment with laccase also decreased the mean germination time by about 1 day as compared to untreated controls. These results show that germinability inhibition due to OMW can be reduced effectively using fungal laccase, suggesting that phenols are the main determinants of its phytotoxicity.

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Arsenic (As) ranks among the priority metal(loid)s that are of public health concern. In the environment, arsenic is present in different forms, organic or inorganic, featured by various toxicity levels. Bacteria have developed different strategies to deal with this toxicity involving different resistance genetic determinants. Bacterial strains of Rhodococcus genus, and more in general Actinobacteria phylum, have the ability to cope with high concentrations of toxic metalloids, although little is known on the molecular and genetic bases of these metabolic features. Here we show that Rhodococcus aetherivorans BCP1, an extremophilic actinobacterial strain able to tolerate high concentrations of organic solvents and toxic metalloids, can grow in the presence of high concentrations of As(V) (up to 240 mM) under aerobic growth conditions using glucose as sole carbon and energy source. Notably, BCP1 cells improved their growth performance as well as their capacity of reducing As(V) into As(III) when the concentration of As(V) is within 30–100 mM As(V). Genomic analysis of BCP1 compared to other actinobacterial strains revealed the presence of three gene clusters responsible for organic and inorganic arsenic resistance. In particular, two adjacent and divergently oriented ars gene clusters include three arsenate reductase genes ( arsC 1/2/3) involved in resistance mechanisms against As(V). A sequence similarity network (SSN) and phylogenetic analysis of these arsenate reductase genes indicated that two of them (ArsC2/3) are functionally related to thioredoxin (Trx)/thioredoxin reductase (TrxR)-dependent class and one of them (ArsC1) to the mycothiol (MSH)/mycoredoxin (Mrx)-dependent class. A targeted transcriptomic analysis performed by RT-qPCR indicated that the arsenate reductase genes as well as other genes included in the ars gene cluster (possible regulator gene, arsR , and arsenite extrusion genes, arsA, acr3 , and arsD ) are transcriptionally induced when BCP1 cells were exposed to As(V) supplied at two different sub-lethal concentrations. This work provides for the first time insights into the arsenic resistance mechanisms of a Rhodococcus strain, revealing some of the unique metabolic requirements for the environmental persistence of this bacterial genus and its possible use in bioremediation procedures of toxic metal contaminated sites.

Consumer demand for vegetables of fortified mineral and bioactive content is on the rise, driven by the growing interest of society in fresh products of premium nutritional and functional quality. Biofortification of leafy vegetables with essential micronutrients such as iron (Fe) is an efficient means to address the human micronutrient deficiency known as hidden hunger. Morphometric analysis, lipophilic and hydrophilic antioxidant capacities of green and red butterhead lettuce cultivars in response to Fe concentration in the nutrient solution (0.015 control, 0.5, 1.0 or 2.0 mM Fe) were assessed. The experiment was carried out in a controlled-environment growth chamber using a closed soilless system (nutrient film technique). The percentage of yield reduction in comparison to the control treatment was 5.7%, 13.5% and 25.3% at 0.5, 1.0 and 2.0 mM Fe, respectively. Irrespective of the cultivar, the addition of 1.0 mM or 2.0 mM Fe in the nutrient solution induced an increase in the Fe concentration of lettuce leaves by 20.5% and 53.7%, respectively. No significant effects of Fe application on phenolic acids and carotenoid profiles were observed in green Salanova. Increasing Fe concentration in the nutrient solution to 0.5 mM triggered a spike in chlorogenic acid and total phenolics in red Salanova lettuce by 110.1% and 29.1% compared with the control treatment, respectively; moreover, higher accumulation of caffeoyl meso tartaric phenolic acid by 31.4% at 1.0 mM Fe and of carotenoids violaxanthin, neoxanthin and β-carotene by 37.0% at 2.0 mM Fe were also observed in red Salanova compared with the control (0.015 mM Fe) treatment. Red Salanova exhibited higher yield, P and K contents, ascorbic acid, phenolic acids and carotenoid compounds than green Salanova. The wok shows how nutrient solution management in soilless culture could serve as effective cultural practices for producing Fe-enriched lettuce of premium quality, notwithstanding cultivar selection being a critical underlying factor for obtaining high quality products.

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