Mechanisms of tolerance and high degradation capacity of the herbicide mesotrione by Escherichia coli strain DH5-α.

The intensive use of agrochemicals has played an important role in increasing agricultural production. One of the impacts of agrochemical use has been changes in population structure of soil microbiota. The aim of this work was to analyze the adaptive strategies that bacteria use to overcome oxidative stress caused by mesotrione, which inhibits 4-hydroxyphenylpyruvate dioxygenase. We also examined antioxidative stress systems, saturation changes of lipid membranes, and the capacity of bacteria to degrade mesotrione. Escherichia coli DH5-á was chosen as a non-environmental strain, which is already a model bacterium for studying metabolism and adaptation. The results showed that this bacterium was able to tolerate high doses of the herbicide (10× field rate), and completely degraded mesotrione after 3 h of exposure, as determined by a High Performance Liquid Chromatography. Growth rates in the presence of mesotrione were lower than in the control, prior to the period of degradation, showing toxic effects of this herbicide on bacterial cells. Changes in the saturation of the membrane lipids reduced the damage caused by reactive oxygen species and possibly hindered the entry of xenobiotics in the cell, while activating glutathione-S-transferase enzyme in the antioxidant system and in the metabolizing process of the herbicide. Considering that E. coli DH5-α is a non-environmental strain and it had no previous contact with mesotrione, the defense system found in this strain could be considered non-specific. This bacterium system response may be a general adaptation mechanism by which bacterial strains resist to damage from the presence of herbicides in agricultural soils

MDA levels in <i>E. coli</i> DH5-α in MM, and in the MMM, -C and -CM treatments, during periods of 30 min., 3 h and 6 h, respectively.

<p>LSD = 0.18 for all pairwise comparison.</p

GST activity of <i>E. coli</i> DH5-α in MM, and in the MMM, -C, and -CM treatments, at periods of 30 min. 3 h and 6 h, respectively.

<p>LSD = 0.000398 for all pairwise comparison.</p

Non-denaturing-PAGE for SOD activity.

<p>Patterns presented by <i>E. coli</i> DH5-α in MM (lines 1, 2 and 3) and in the MMM (lines 4, 5 and 6), -C (lines 7, 8 and 9), and -CM (lines 10, 11 and 12) treatments, at periods of 30 min., 3 h and 6 h, respectively.</p

Cellular viability of <i>E. coli</i> DH5-α in MM and in the MMM, -C and –CM treatments, during the periods of 30 min., 3 h and 6 h.

<p>LSD = 0.13 for all pairwise comparison.</p

Degradation kinetics of mesotrione mediated by <i>E. coli</i> DH5-α.

<p>MMM (mineral medium with mesotrione), -CM (mineral medium without carbon, with mesotrione), negative control (MMM without <i>E. coli</i> DH5-α) and boiled cells.</p

Molecular structure of mesotrione.

<p>Molecular structure of mesotrione.</p

<i>Bacillus megaterium</i> strains derived from water and soil exhibit differential responses to the herbicide mesotrione

<div><p>The intense use of herbicides for weed control in agriculture causes selection pressure on soil microbiota and water ecosystems, possibly resulting in changes to microbial processes, such as biogeochemical cycles. These xenobiotics may increase the production of reactive oxygen species and consequently affect the survival of microorganisms, which need to develop strategies to adapt to these conditions and maintain their ecological functionality. This study analyzed the adaptive responses of bacterial isolates belonging to the same species, originating from two different environments (water and soil), and subjected to selection pressure by herbicides. The effects of herbicide Callisto and its active ingredient, mesotrione, induced different adaptation strategies on the cellular, enzymatic, and structural systems of two <i>Bacillus megaterium</i> isolates obtained from these environments. The lipid saturation patterns observed may have affected membrane permeability in response to this herbicide. Moreover, this may have led to different levels of responses involving superoxide dismutase and catalase activities, and enzyme polymorphisms. Due to these response systems, the strain isolated from water exhibited higher growth rates than did the soil strain, in evaluations made in oligotrophic culture media, which would be more like that found in semi-pristine aquatic environments. The influence of the intracellular oxidizing environments, which changed the mode of degradation of mesotrione in our experimental model and produced different metabolites, can also be observed in soil and water at sites related to agriculture. Since the different metabolites may present different levels of toxicity, we suggest that this fact should be considered in studies on the fate of agrochemicals in different environments.</p></div

Infrared spectrum profile of the lipid extract obtained after incubation on MM (Panel 1); MMM (Panel 2) and MMC (Panel 3) of <i>B</i>. <i>megaterium</i> CCT 7729, (A) 3 h, (B) 14 h and <i>B</i>. <i>megaterium</i> CCT 7730, (C) 3 h (D) 14 h (FTIR analysis with scans from 4,000 cm^-1 to 400 cm^-1).

<p>Bands 1 e 2 correspond to (CH₂); 3 to (CH₃₎; 4 to (C = O); 5 to (-C = C); 6 to (CH₂); 7 to (CH₃); 8 to (C-O) and to 9 (CH).</p