Abstract
Key message
Mid-bicellular pollen vegetative cells in tobacco escape from G1 arrest and proceed to the G1/S transition towards androgenesis within 1 day under glutamine starvation conditions in vitro.
Abstract
In the Nicotiana tabacum pollen culture system, immature pollen grains at the mid-bicellular stage can mature in the presence of glutamine; however, if glutamine is absent, they deviate from their native cell fate in a few days. The glutamine-starved pollen grains cannot undergo maturation, even when supplied with glutamine later. Instead, they undergo cell division towards androgenesis slowly within 10 days in a medium containing appropriate nutrients. During the culture period, they ought to escape from G1 arrest to proceed into S phase as the primary step towards androgenesis. However, this event has not been experimentally confirmed. Here, we demonstrated that the pollen vegetative cells proceeded to the G1/S transition within approximately 15–36 h after the start of culture. These results were obtained by analyzing transgenic pollen possessing a fusion gene encoding nuclear-localizing GFP under the control of an E2F motif-containing promoter isolated from a gene encoding one of DNA replication licensing factors. Observations using a 5-ethynyl-2′-deoxyuridine DNA labeling and detection technique uncovered that the G1/S transition was soon followed by S phase. These hallmarks of vegetative cells undergoing dedifferentiation give us new insights into upstream events causing the G1/S transition and also provide a novel strategy to increase the frequency of the androgenic response in tobacco and other species, including recalcitrants.
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Acknowledgments
We thank the students in our laboratory at the time when this research was conducted, K. Kohra, K. Goto, A. Kawamoto, W. Takahashi, and Y. Hori, for their efforts towards completing this study.
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The authors have no competing financial interests to declare.
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Communicated by Fumihiko Sato.
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Kyo, M., Nagano, A., Yamaji, N. et al. Timing of the G1/S transition in tobacco pollen vegetative cells as a primary step towards androgenesis in vitro. Plant Cell Rep 33, 1595–1606 (2014). https://doi.org/10.1007/s00299-014-1640-5
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DOI: https://doi.org/10.1007/s00299-014-1640-5