Abstract
Programmed cell death (PCD) processes are essential in the plant embryogenesis. To understand how PCD operates in a developing seed, the dying cells need to be identified in relation to their surviving neighbors. This can be accomplished by the means of in situ visualization of fragmented DNA—a well-known hallmark of PCD. In the developing Scots pine (Pinus sylvestris L.) seed, several tissues die via morphologically different PCD processes during the embryogenesis. Here, we describe the protocols for the characterization of Scots pine seeds at the early and late developmental stages and, further, the localization of nucleic acids and DNA fragmentation by the acridine orange staining and TUNEL (terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick end labeling) assay in the dying seed tissues.
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Acknowledgments
The authors wish to warmly thank Dr. Anne Jokela, Dr. Riina Muilu-Mäkelä, M.Sc. Mira Sääskilahti, M.Sc. Johanna Kestilä, and Ms. Eeva Pihlajaviita for their significant contribution to the developing and optimization of the presented procedures.
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Vuosku, J., Sutela, S. (2020). Microscopical Detection of Cell Death Processes During Scots Pine Zygotic Embryogenesis. In: Bayer, M. (eds) Plant Embryogenesis. Methods in Molecular Biology, vol 2122. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0342-0_16
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DOI: https://doi.org/10.1007/978-1-0716-0342-0_16
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Publisher Name: Humana, New York, NY
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