The bleaching induced by the phenylpyridazinone herbicide Metflurazon in Chlorella is a metabolic and reversible process

The treatment of synchronized cells of the green alga Chlorella fusca under photoautotrophic conditions with metflurazon (SAN H 6706; 4-chloro-5-dimethylamino-2-(α,α,α-trifluoro-m-tolyl-3(2H)-pyridazinone)) induces a bleaching process and results in white-appearing cells. This process of bleaching was followed by quantitative analysis of cell growth and cell division, photosynthetic oxygen evolution, respiratory oxygen consumption, and of pigment pattern at 0, 6, 12, 18, 24, 48, and 72 hr after incubation with different concentrations of metflurazon. Increasing concentrations of metflurazon gradually affected cell growth of Chlorella measured as increase in cell diameter. Cell division was inhibited completely with 1, 10, and 100 μM metflurazon. Photosynthetic oxygen evolution and respiratory oxygen consumption were not inhibited by 1 μM metflurazon during the first 6 hr; after this time a gradually increased inhibition was observed. Both parameters were inhibited by 100 μM metflurazon immediately after herbicide addition. A detailed analysis of the pigment content during the bleaching process revealed that: (a) The bleaching of Chlorella cells by metflurazon is not a simple photochemical process like the photobleaching of boiled cells, but is directed by the active metabolism of Chlorella itself. (b) The bleaching process is characterized by two phases: an accumulation of pigments followed by their degradation. The accumulation phase extends to 6 hr after herbicide addition. (c) During the accumulation phase, chlorophyll is accumulated to 380 and 106% in cells treated with 1 and 100 μM metflurazon, respectively, compared to the initial pigment content. The breakdown of chlorophyll, however, during the degradation phase is 5 times faster in the 1 μM treatment than in the 100 μM treatment. This difference resulted in the faster appearance of white cells with the low metflurazon concentration. (d) During the accumulation phase in the 1 μM treatment, the biosynthesis of chlorophylls, xanthophylls, and carotenes is inhibited by 56, 74, and 78%, respectively, when compared to a nontreated control. When related to the initial amounts, chlorophylls, xanthophylls, and carotenes are accumulated to 380, 230, and 153%, respectively. However, the synthesis of violaxanthin is specifically inhibited, followed by α-carotene. During the degradation phase, violaxanthin and α-carotene again, are the most rapidly disappearing pigments. Continuous culturing of white Chlorella cells resulted in a regeneration to green cells after 96, 240, 384 hr for 1, 10, and 100 μM metflurazone, respectively. The bleaching of Chlorella by metflurazon is evidently dependent on a functioning metabolism and is itself a regulated disassembly of the photosynthetic apparatus, which is reversible and not lethal.