Influence of mixotrophy on cell cycle phase duration and correlation of karlotoxin synthesis with light and G1 phase in Karlodinium veneficum

Karlodinium veneficum forms fish killing blooms in estuaries worldwide. The toxicity of these blooms is variable and thought to be connected to bloom stage and in situ growth rates. Methods for measuring in situ growth rates rely on the assumption that cell cycle progression is phased to the diel photocycle, which is true for phototrophically-growing K. veneficum cultures where G1 phase occurs during light and S and G2 + M phases occur during darkness. However, K. veneficum is a facultative mixotroph that also phagocytizes microalgal prey, and the effects of this mixotrophy on its cell cycle synchrony are unknown. Furthermore, toxicity in laboratory cultures is inversely related to growth rate and is light dependent, suggesting synchrony between the cell cycle (G1 phase) and karlotoxin synthesis. To test this, the cell cycle phase distribution and cellular toxin content for phototrophic and mixotrophic cultures of K. veneficum were monitored hourly for a full diel cycle. The results demonstrated that mixotrophic cultures maintained a synchronized cell cycle, despite increased growth rates. The faster growth rates were attributed to a shortened duration of G1 phase in mixotrophic cultures compared to phototrophic cultures (30.8 ± 9.2 h vs 69.4 ± 21.5 h, respectively). Meanwhile, toxin production was observed only during light hours, consistent with synthesis initiating with the photorespiratory byproduct glycolate. Cellular toxin content had a significant positive correlation with the percentage of G1 phase cells and a significant negative correlation with the percentage of S phase cells. These results indicate a clear role in mixotrophy increasing growth rates of K. veneficum and of the diel photocycle in synchronizing the cell and karlotoxin synthetic cycles.