Abstract
Cyanidioschyzon merolae 10D was originally isolated from a mixture of hot spring water sampled in Naples, Italy. Currently, this strain is available in the Microbial Culture Collection at the National Institute of Environmental Studies in Japan. The strain has been cultured in 2× Allen’s medium or its derivatives. The optimal growth conditions for this strain are as follows: pH 2.5, 42 °C, and ~100 μmol photons m−2 s−1, allowing the cell density to reach ~5 × 108 cells mL−1. C. merolae can also grow slowly at >20 °C. We generally store stock cultures at room temperature or at 30 °C under a low light condition (~20 μmol photons m−2 s−1) or as frozen stock in liquid nitrogen. Cell cycle progression can be synchronized by subjecting the culture to a 12-h light/12-h dark cycle. In addition, cells can be arrested at the S or M phases by adding relevant inhibitors. The shapes of cells and chloroplasts are clearly observed by phase-contrast or differential interference contrast microscopy. Because C. merolae lacks a cell wall, cellular contents (e.g., DNA, RNA, and proteins) are easily extracted.
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Acknowledgments
We thank Dr. Kuroiwa (Japan Women’s University), Drs. Tanaka and Imamura (Tokyo Institute of Technology), Dr. Misumi (Yamaguchi University), Dr. Yoshikawa (Tokyo University of Agriculture), and their lab members and members of Miyagishima lab in National Institute of Genetics for providing information on C. merolae experiments. Our study was partly supported by Japan Society for the Promotion of Science Grant-in-Aid for Scientific Research 25251039 (to S.M.) and by the Core Research for Evolutional Science and Technology Program of the Japan Science and Technology Agency (to S.M.).
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Miyagishima, S., Wei, J.L. (2017). Procedures for Cultivation, Observation, and Conventional Experiments in Cyanidioschyzon merolae . In: Kuroiwa, T., et al. Cyanidioschyzon merolae. Springer, Singapore. https://doi.org/10.1007/978-981-10-6101-1_3
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DOI: https://doi.org/10.1007/978-981-10-6101-1_3
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