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Measuring Endoreduplication by Flow Cytometry of Isolated Tuber Protoplasts
Published on: March 9, 2018
Ethylene stimulates endoreduplication but inhibits cytokinesis in cucumber hypocotyl epidermis
Haruka Dan1, Hidemasa Imaseki, Geoffrey O Wasteneys
1Department of Biology, International Christian University, Mitaka, Tokyo 181-8585 Japan.
Abstract:
The effects of ethylene on cell division are generally considered inhibitory. In this study, we demonstrate that transient ethylene exposure, while suppressing cytokinesis, stimulates DNA synthesis. We monitored DNA synthesis and cytokinesis in the epidermis of cucumber (Cucumis sativus) hypocotyls, an organ whose post-germination development involves strictly limited cell division. During exposure to ethylene, DNA synthesis, assessed by the incorporation of the thymidine homolog 5-bromo-2'-deoxyuridine, was detected in 20% of the epidermal cells, whereas DNA synthesis was nearly undetectable in normal air. Cytofluorometric analysis of nuclei in affected cells showed an up to 8-fold increase in DNA content. During this time, new cell plate formation was not detected. However, shortly after ethylene was removed, DNA content was rapidly restored to 2C (diploid) levels in all cells, and new cell plate formation dramatically increased. These results demonstrate that ethylene promotes DNA synthesis and its endoreduplication but inhibits cytokinesis, thereby maintaining some cells in G2 phase.
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