Chinese Bulletin of Botany ›› 2015, Vol. 50 ›› Issue (3): 363-371.DOI: 10.3724/SP.J.1259.2015.00363

• TECHNIQUES AND METHODS • Previous Articles     Next Articles

Highly Synchronized Chlamydomonas reinhardtii Cells by Dark-shifting a Continuous-light Culture Combined with Drugs

Weizhong Liu1*, Jieru Wang1, Yong Hu2   

  1. 1College of Life Sciences, Shanxi Normal University, Linfen 041004, China;
    2.College of Life Sciences, Capital Normal University, Beijing 100048, China
  • Received:2014-12-25 Revised:2015-03-19 Online:2015-05-01 Published:2015-04-08

Abstract: Chlamydomonas reinhardtii, a unicellular eukaryotic green alga, has been a powerful model system for understanding several fundamental biological processes such as photosynthesis and plant cell-cycle regulation. An artificial cell-cycle synchronization system is necessary for these studies. Here, we investigated a new method for synchronizing Chlamydomonas cell-wall-less cells, with which a high level of synchrony can be achieved. The degree of synchrony was analysed by flow cytometry. The cells are grown asynchronously in continuous light, then grown for 24 h in the dark (dark-shifting a continuous-light culture). At the end of the dark period, most cells (>80%) are arrested in the G1 phase. If necessary, an S-phase blocking agent can be used to treat the dark-shifted cells. When the dark-shifted cells are treated with M-phase blocking agent alone or with the S-phase and M-phase blocking agent sequentially, a cell population with more than 80% of cells at the M phase can be achieved. A specific synchronous procedure needs to be selected according to the properties of a certain Chlamydomonas strain (especially certain mutants) and the requirement of research.