Partial molecular analysis of the psbA gene in Euglena gracilis mutants exhibiting resistance to DCMU and atrazine.

Mutations conferring herbicide resistance have been detected in two new strains (ZR250 and ZR480) of Euglena gracilis Z by partial gene cloning and sequencing. These mutants were originally derived from Z cells grown in medium containing progressively increasing concentrations of DCMU. Each of these strains have been characterized by measuring their growth kinetics, O2 evolution, and resistance to DCMU and atrazine. Partial sequences of the psbA gene of these strains were compared to those published for strains Z and ZR25. The ZR250 and ZR480 strains were found to be double mutants. Besides the expected mutation S265A, they showed an additional point mutation at codon 219 (equivalent to codon 218 of other organisms). This mutation results in leucine being substituted by phenylalanine. For each of the ZR strains, two growth conditions (with or without DCMU in the medium) have been compared. The presence of the second mutation (at codon 219) leads to notable increase (20-fold) in resistance to DCMU, whereas the resistance to atrazine is only 2-fold. The presence of DCMU, as the selective agent, was responsible for an enhanced herbicide resistance, irrespective of the concentrations used. Substantial modifications in the rate of cell growth and O2 yields were observed when the maximal concentration (480 microM) of DCMU was used. These modifications were reversible on withdrawal of the DCMU. Thus the reversible adaptative modifications also adds to the mutational effect observed.

Citrulline-malate effect on microsome phospholipids and cytochrome P450 in Euglena grown with ethanol.

This study indicates for the first time the presence of cytochrome P450 in the microsomes of Euglena grown in lactate medium and substantiates the use of Euglena as a hepatic cell model. Similar effects of ethanol on Euglena and on rat hepatic microsomes were demonstrated: (i) decrements in the quantities of FA per milligram of proteins; (ii) increases in the proportions of PE; (iii) decreases in the proportions of PC; and (iv) production of cytochrome P450, degraded in P420. The citrulline-malate reestablishes in the microsomes the phospholipid environment and the cytochrome P450 concentration. These findings illustrate that the complex acts on the lipid peroxidation via the changes in cytochrome P450 activity.

Effects of a first exposure to ethanol on the compositions of neutral and polar lipids in Euglena gracilis Z, taken as a hepatic cell model: equilibration by citrulline-malate.

In comparison to the lipid composition of Euglena cells fed with lactate, a first exposure of the cells to ethanol favors the production of neutral lipids containing mainly unsaturated fatty acids. The ethanol diminishes drastically the proportion of PC and weakly that of PE. In contrast, it increases slightly the proportion of DPG. The ethanol induces important changes in the fatty acid distributions of each lipid class, suggesting modifications of the elongation-desaturation system. On the one hand the proportion of unsaturated fatty acids is increased and, on the other hand, the last double bond is predominantly situated in the delta 6 position in place of delta 3. The addition of the complex citrulline-malate corrects most of these changes.