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Photoperiods have an important impact on macroalgae living in the intertidal zone. Ocean acidification also influences the physiology of macroalgae. However, little is known about the interaction between ocean acidification and photoperiod on macroalgae. In this study, a green alga Ulva linza was cultured under three different photoperiods (L:D = 8:16, 12:12, 16:8) and two different CO2 levels (LC, 400 ppm; HC, 1,000 ppm) to investigate their responses. The results showed that relative growth rate of U. linza increased with extended light periods under LC but decreased at HC when exposed to the longest light period of 16 h compared to 12 h. Higher CO2 levels enhanced the relative growth rate at a L:D of 8:16, had no effect at 12:12 but reduced RGR at 16:8. At LC, the L:D of 16:8 significantly stimulated maximum quantum yield (Yield). Higher CO2 levels enhanced Yield at L:D of 12:12 and 8:16, had negative effect at 16:8. Non-photochemical quenching (NPQ) increased with increasing light period. High CO2 levels did not affect respiration rate during shorter light periods but enhanced it at a light period of 16 h. Longer light periods had negative effects on Chl a and Chl b content, and high CO2 level also inhibited the synthesis of these pigments. Our data demonstrate the interactive effects of CO2 and photoperiod on the physiological characteristics of the green tide macroalga Ulva linza and indicate that future ocean acidification may hinder the stimulatory effect of long light periods on growth of Ulva species.