期刊
BMC BIOLOGY
卷 15, 期 -, 页码 -出版社
BMC
DOI: 10.1186/s12915-016-0344-4
关键词
Cyanobacteria; Microfluidics; Single-cell imaging; Light-dark cycles; Cell-size homeostasis; Circadian clock; Photosynthesis
类别
资金
- NSF [EF-1038697, MCB-1149328, MCB-1331151]
- Bio-X Interdisciplinary Initiatives Program seed grant
- National Academies Keck Future Initiatives seed grant
Background: Cyanobacteria are important agents in global carbon and nitrogen cycling and hold great promise for biotechnological applications. Model organisms such as Synechocystis sp. and Synechococcus sp. have advanced our understanding of photosynthetic capacity and circadian behavior, mostly using population-level measurements in which the behavior of individuals cannot be monitored. Synechocystis sp. cells are small and divide slowly, requiring long-term experiments to track single cells. Thus, the cumulative effects of drift over long periods can cause difficulties in monitoring and quantifying cell growth and division dynamics. Results: To overcome this challenge, we enhanced a microfluidic cell-culture device and developed an image analysis pipeline for robust lineage reconstruction. This allowed simultaneous tracking of many cells over multiple generations, and revealed that cells expand exponentially throughout their cell cycle. Generation times were highly correlated for sister cells, but not between mother and daughter cells. Relationships between birth size, division size, and generation time indicated that cell-size control was inconsistent with the sizer rule, where division timing is based on cell size, or the timer rule, where division occurs after a fixed time interval. Instead, single cell growth statistics were most consistent with the adder rule, in which division occurs after a constant increment in cell volume. Cells exposed to light-dark cycles exhibited growth and division only during the light period; dark phases pause but do not disrupt cell-cycle control. Conclusions: Our analyses revealed that the adder model can explain both the growth-related statistics of single Synechocystis cells and the correlation between sister cell generation times. We also observed rapid phenotypic response to light-dark transitions at the single cell level, highlighting the critical role of light in cyanobacterial cell-cycle control. Our findings suggest that by monitoring the growth kinetics of individual cells we can build testable models of circadian control of the cell cycle in cyanobacteria.
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