Investigation of the agroecological applications of olive mill wastewater fractions from the ultrafiltration-nanofiltration process.

Agricultural applications of olive mill wastewater (OMW) represent a critical challenge, consistent with waste recycling and the trend towards a more sustainable pattern of agriculture. In this context, an integrated study on the agroecological applications of OMW from the ultrafiltration (UF) - nanofiltration (NF) process was carried out. This process generated three fractions: UF retentate and NF permeate, depleted in salts and phenolic compounds, were studied for their fertilization and irrigation potential, while NF retentate, enriched in these elements, was studied for its potential as a bioherbicide. The phytotoxicity of the NF retentate fraction on two crops (maize and flax) was evaluated on seedlings growth and chloroplast pigments content. In addition, the induced defense responses in maize and flax seedlings were investigated by measuring two parameters: the activity of the detoxification enzyme glutathione-S-transferase (GST) and the concentration of polyphenols, as a component of the antioxidant defense strategy in plants. Biomass, height, and chloroplast pigments content decreased progressively with increasing NF retentate concentration. Conversely, an increase in GST activity and polyphenol concentration was observed. These results highlighted the ability of OMW to induce an oxidative stress on maize and flax seedlings, triggering a defense response through GST and phenolic compounds. On the other hand, in vitro tests on the phytotoxicity of the NF retentate fraction on the common weed Sinapis arvensis were carried out. No germination was observed even with the lowest dilution applied, thus establishing the first data about the selectivity of potential OMW-derived bioherbicides. On the other hand, UF retentate and NF permeate treatments led to a significant increase in maize growth: these fractions could then be considered as a promising organic fertilizer for degraded agricultural soils, as well as an alternative water source for crops irrigation.

Engineering herbicide metabolism in tobacco and Arabidopsis with CYP76B1, a cytochrome P450 enzyme from Jerusalem artichoke.

The Jerusalem artichoke (Helianthus tuberosus) xenobiotic inducible cytochrome P450, CYP76B1, catalyzes rapid oxidative dealkylation of various phenylurea herbicides to yield nonphytotoxic metabolites. We have found that increased herbicide metabolism and tolerance can be achieved by ectopic constitutive expression of CYP76B1 in tobacco (Nicotiana tabacum) and Arabidopsis. Transformation with CYP76B1 conferred on tobacco and Arabidopsis a 20-fold increase in tolerance to linuron, a compound detoxified by a single dealkylation, and a 10-fold increase in tolerance to isoproturon or chlortoluron, which need successive catalytic steps for detoxification. Two constructs for expression of translational fusions of CYP76B1 with P450 reductase were prepared to test if they would yield even greater herbicide tolerance. Plants expressing these constructs had lower herbicide tolerance than CYP76B1 alone, which is apparently a consequence of reduced stability of the fusion proteins. In all cases, increased herbicide tolerance results from more extensive metabolism, as demonstrated with exogenously fed phenylurea. Beside increased herbicide tolerance, expression of CYP76B1 has no other visible phenotype in the transgenic plants. Our data indicate that CYP76B1 can function as a selectable marker for plant transformation, allowing efficient selection in vitro and in soil-grown plants. Plants expressing CYP76B1 may also be a potential tool for phytoremediation of contaminated sites.