Efficacy of an organically modified bentonite to adsorb 2,4-dichlorophenoxyacetic acid (2,4-D) and prevent its phytotoxicity.

The herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) is widely used due to it selective action, and preferential control of dicotyledonous weeds affecting cereal crops. Physiological responses of sensitive dicotyledonous plants to 2,4-D include growth retardation, senescence, and cell death. Due to soil and water contamination by agricultural practices, 2,4-D constitutes a potential risk to non-target plant species. In this work, the potential advantage of using organic modified bentonite (Bent) to adsorb 2,4-D and therefore mitigate damage produced by this herbicide on sensitive not-target vegetable species was investigated. Dodecylamine (DDA) was used as an organic modifier to change the hydrophilic nature of Bent into an organophilic matrix. The adsorption performances of 2,4-D by Bent-DDA were analyzed. The maximum adsorptions of 2,4-D (22.1 mg/L) from aqueous solution containing 1.0 or 2.5 mg/mL Bent-DDA were 40 and 80 %, respectively. The physical interaction of Bent-DDA with 2,4-D was characterized by Wide Angle X-ray Scattering (WAXS) and thermogravimetric analysis (TGA). The biological functionality of Bent-DDA matrix as 2,4-D adsorbent was tested in a bioassay in the Arabidopsis thaliana plant model system. The primary root growth of Arabidopsis seedlings is strongly inhibited by low concentrations of 2,4-D. Arabidopsis seedlings submitted to Bent-DDA pre-treated herbicide aqueous solution showed similar root growth than 2,4-D non-treated seedlings. Finally, the ability of Bent-DDA to prevent 2,4-D phytotoxicity was exploratory investigated in lettuce plants. Lettuce plants pre-treated with 20 µg/mL Bent-DDA showed reduced sensitivity to 2,4-D including an increment on chlorophyll content and biomass compared with non-treated plants. Our findings revealed a promising scenario for the application of Bent-DDA as an effective adsorbent of 2,4-D at productive scale.

Evidence on antimicrobial properties and mode of action of a chitosan obtained from crustacean exoskeletons on Pseudomonas syringae pv. tomato DC3000.

Pseudomonas syringae pv. tomato DC3000 (Pto DC3000) causes bacterial speck of tomato, a widely spread disease that causes significant economical losses worldwide. It is representative of many bacterial plant diseases for which effective controls are still needed. Despite the antimicrobial properties of chitosan has been previously described in phytopathogenic fungi, its action on bacteria is still poorly explored. In this work, we report that the chitosan isolated from shrimp exoskeletons (70 kDa and 78 % deacetylation degree) exerts cell damage on Pto DC3000. Chitosan inhibited Pto DC3000 bacterial growth depending on its concentration, medium-pH, and presence of metal ion (Mg(+2)). Biochemical and cellular changes resulting in cell aggregation and impaired bacterial growth were also viewed. In vivo studies using fluorescent probes showed cell aggregation, increase in membrane permeability, and cell death, suggesting the chitosan antibacterial activity is due to its interaction as a polycation with Pto DC3000 membranes. Transmission electron microscopic analysis revealed that chitosan also caused morphological changes and damage in bacterial surfaces. Also, the disease incidence in tomato inoculated with Pto DC3000 was significantly reduced in chitosan pretreated seedlings, revealing a promising action of chitosan as nontoxic biopesticide in tomato plants. Indeed, a wider comprehensive knowledge of the mechanism of action of chitosan in phytopathogenic bacterial cells will increase the chances of its successful application to the control of spread disease in plants.

Multicatalytic proteinase in fish muscle.

A partially active and a latent form of multicatalytic protease (MCP) were isolated from fish skeletal muscle. Both forms were inactive against protein substrates, but their activity against peptide substrates differed in one order of magnitude. The chymotrypsin-like activity of the partially active form was moderately stimulated by fatty acids and SDS, whereas its trypsin-like activity was inhibited by the same reagents. In contrast, both activities of the latent form were strongly stimulated by SDS. The chymotrypsin-like activity of the latent form was also stimulated by heating or high urea concentrations, whereas its trypsin-like activity did not change or was inhibited respectively by these treatments. These activation effects were irreversible. Pre-treatment of the latent form with SDS or urea in the absence of substrate led to its irreversible inactivation, whereas activation by pre-heating occurred in the presence or absence of substrate. These results suggest that MCP can exist in several active states with distinct properties. Studies on the distribution of MCP in fish tissues showed a much higher level of the enzyme in gonads than in any other tissue, suggesting a role of MCP in development.