The engineered microalgae were cultivated in fresh medium for several times to avoid the influence of antibiotic zeocin. Thereafter, engineered and WT cells were harvested at 3000g for 10 min and resuspended in fresh f/2 medium without antibiotic, and algal cell concentration was made up to 1 × 106 cells/ml. Growth curve was determined by counting cells with a Brightline hemocytometer under a light microscope every day. Both overexpressed and WT cells on 11th day were incubated in dark for at least 40 min, and thereafter, the chlorophyll fluorescence (Fv/Fm; the maximum photochemical efficiency of PSII) was measured with a TD-700 fluorometer (Turner Design, USA) (17).

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