Flow cytometry and 5‐ethynyl‐2′‐deoxyuridine (EdU) labeling to detect the cell cycle dynamics of Phaeodactylum tricornutum under light

作者：Ting Zhang, Jingyi Huang, Zhixia Zhang, Jie Lv, Dongqun Zhang, Renwei Qing, Liqiong Lan · 发表于：Journal of Phycology · 年份：2022 · DOI：10.1111/jpy.13250 · 被引用次数：6 · 研究领域：Algal biology and biofuel production、Protist diversity and phylogeny、Diatoms and Algae Research

Cell cycle studies in plants and algae are highly dependent on reliable methods for detecting cellular DNA replication. With its short growth cycle and ease of genetic transformation, Phaeodactylum tricornutum is an important model organism for the study of pennate diatoms. Here we explored two different methods to detect the cell cycle of P. tricornutum, one using SYBR-green I to via flow cytometry, and the other using EdU labeling to observe cell cycle changes under fluorescence microscopy. Both EdU labeling fluorescence microscopy and SYBR-green I staining flow cytometry accurately indicated that the cells of P. tricornutum enter the G2/M phase after 12 h of light exposure. The results indicate that SYBR Green I was an adequate detection method for nuclear DNA quantitation in cells of P. tricornutum using a flow cytometer and without RNase A treatment. In addition, EdU can be applied to P. tricornutum to reliably detect cell proliferation. Besides, Mg-ProtoIX was able to reverse the cell cycle division inhibition of P. tricornutum and allow the nuclear DNA replication to proceed normally. Taken together, the photoperiodic division time point was clearly identified, which sheds light on the regulation of cell division mechanism in P. tricornutum.