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To examine the sulfate assimilation and reduction process and the regulation of illumination, diatom Phaeodactylum tricornutum and dinoflagellate Amphidinium carterae were selected for continuous simulation incubation under different photon flux densities (PFDs) (54, 108 and 162 μmol photons m−2 s−1), and concentration variations of related sulfur compounds sulfate, dimethylsulfoniopropionate (DMSP), dimethylsulfide (DMS) and acrylic acid (AA) in the culture system were observed. The optimal PFD for the growth of two microalgae was 108 μmol photons m−2 s−1. However, the maximum sulfate absorption occurred at 162 μmol photons m−2 s−1 for P. tricornutum and at 54 μmol photons m−2 s−1 for A. carterae. With the increase of PFD, the release of DMSP by P. tricornutum decreased while A. carterae increased. The largest release amount of DMS was 0.59 ± 0.05 fmol cells−1 for P. tricornutum and 2.61 ± 0.89 fmol cells−1 for A. carterae under their optimum growth light condition. The sulfate uptake of P. tricornutum was inhibited by the addition of amino acids, cysteine had a greater inhibitory effect than methionine, and the absorption process was controlled by light. The intermediate products of sulfur metabolism had an up–control effect on the sulfate uptake process of P. tricornutum. However, the addition of amino acids had no obvious effect on the sulfate absorption of A. carterae. • Illumination regulated sulfate assimilation of P. tricornutum and A. carterae. • 108 μmol photons m−2 s−1 was the optimum light condition for the algal growth. • Related sulfur compounds concentrations varied under different illumination. • Sulfur metabolic intermediates inhibited the sulfate uptake of P. tricornutum. • Sulfur metabolic intermediates had no obvious regulative law on A. carterae. [ABSTRACT FROM AUTHOR]