Chromochloris zofingiensis (Chlorophyceae) Divides by Consecutive Multiple Fission Cell-Cycle under Batch and Continuous Cultivation

SIMPLE SUMMARY: Microalgae are plant-like micro-organisms naturally found in fresh and marine water environments, inhabiting a vast range of ecosystems. They capture light energy through photosynthesis and convert low energy inorganic compounds (carbon dioxide and water) into high energy complex organic compounds, such as carbohydrates and fats. Chromochloris zofingiensis is a unicellular microalga currently under intensive research, due to its ability to produce high value pharmaceutical and nutritional pigments. Understanding its growth characteristics is crucial for the establishment of an efficient commercial production of those pigments from this alga. Thus, we have developed a method to stain the nucleus of the alga which enabled us to follow the division pattern under commonly used cultivation methods. We found that C. zofingiensis cells conduct consecutive DNA synthesis and divisions of the nucleus to produce 8 or 16 nuclei before it divides into 8 or 16 daughter cells, respectively. Under high light illumination, the whole process lasts several days, through which cells grow during the light period and divide during the dark period. These findings can be assimilated for the development of the biotechnology process for high pigment productivity. ABSTRACT: Several green algae can divide by multiple fission and spontaneously synchronize their cell cycle with the available light regime. The yields that can be obtained from a microalgal culture are directly affected by cell cycle events. Chromochloris zofingiensis is considered as one of the most promising microalgae for biotechnological applications due to its fast growth and the flexible trophic capabilities. It is intensively investigated in the context of bio-commodities production (carotenoids, storage lipids); however, the pattern of cell-cycle events under common cultivation strategies was not yet characterized for C. zofingiensis. In this study, we have employed fluorescence microscopy to characterize the basic cell-cycle dynamics under batch and continuous modes of phototrophic C. zofingiensis cultivation. Staining with SYBR green—applied in DMSO solution—enabled, for the first time, the clear and simple visualization of polynuclear stages in this microalga. Accordingly, we concluded that C. zofingiensis divides by a consecutive pattern of multiple fission, whereby it spontaneously synchronizes growth and cell division according to the available illumination regime. In high-light continuous culture or low-light batch culture, C. zofingiensis cell-cycle was completed within several light-dark (L/D) cycles (14 h/10 h); however, cell divisions were synchronized with the dark periods only in the high-light continuous culture. In both modes of cultivation, daughter cell release was mainly facilitated by division of 8 and 16-polynuclear cells. The results of this study are of both fundamental and applied science significance and are also important for the development of an efficient nuclear transformation system for C. zofingiensis.