Abstract
Wheat zygotes can be mechanically isolated and cultivated to continue their development in vitro. Since each zygote needs to be individually isolated, only relatively few of these cells are available per experiment. To facilitate embryonic growth despite of this limitation, the zygotes are kept within a culture insert placed in a larger dish which itself contains embryogenic pollen cocultivated for continuous medium conditioning. This setup ensures that the two cultures, while being physically separated from one another, can exchange essential intercellular signal molecules passing through the bottom of the insert which is made of a permeable membrane. Thanks to the natural fate of zygotes, which is to form an embryo followed by the generation of a plant, embryogenesis and plant regeneration are achieved at much higher efficiency as compared to other single-cell systems. While the method is largely independent of the genotype, it allows for the nondestructive observation, manipulation, and individual analysis of zygotes and very young embryos.
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Acknowledgment
I would like to thank my colleague Dr. Maia Gurushidze for critically reading the manuscript.
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Kumlehn, J. (2016). Embryogenesis and Plant Regeneration from Isolated Wheat Zygotes. In: Germana, M., Lambardi, M. (eds) In Vitro Embryogenesis in Higher Plants. Methods in Molecular Biology, vol 1359. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3061-6_29
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DOI: https://doi.org/10.1007/978-1-4939-3061-6_29
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3060-9
Online ISBN: 978-1-4939-3061-6
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