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Nuclear fragmentation and DNA degradation during programmed cell death in petals of morning glory (Ipomoea nil)

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Abstract

We studied DNA degradation and nuclear fragmentation during programmed cell death (PCD) in petals of Ipomoea nil (L.) Roth flowers. The DNA degradation, as observed on agarose gels, showed a large increase. Using DAPI, which stains DNA, and flow cytometry for DAPI fluorescence, we found that the number of DNA masses per petal at least doubled. This indicated chromatin fragmentation, either inside or outside the nucleus. Staining with the cationic lipophilic fluoroprobe DiOC6 indicated that each DNA mass had an external membrane. Fluorescence microscopy of the nuclei and DNA masses revealed an initial decrease in diameter together with chromatin condensation. The diameters of these condensed nuclei were about 70% of original. Two populations of nuclear diameter, one with an average diameter about half of the other, were observed at initial stages of nuclear fragmentation. The diameter of the DNA masses then gradually decreased further. The smallest observed DNA masses had a diameter less than 10% of that of the original nucleus. Cycloheximide treatment arrested the cytometrically determined changes in DNA fluorescence, indicating protein synthesis requirement. Ethylene inhibitors (AVG and 1-MCP) had no effect on the cytometrically determined DNA changes, suggesting that these processes are not controlled by endogenous ethylene.

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Abbreviations

AVG:

Aminoethoxyvinyl glycine

CHI:

Cycloheximide

DAPI:

4,6-Diamidino-2-phenylindole

DiOC6 :

3,3′-Dihexyloxacarbocyanine iodide

1-MCP:

1-Methylcyclopropene

PCD:

Programmed cell death

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Author notes

  1. Tetsuya Yamada

    Present address: Graduate School of Agriculture, Tokyo University of Agriculture and Technology, 183-8509, Tokyo, Japan

Authors and Affiliations

  1. Plant Biotechnology Institute, Ibaraki Agricultural Center, 319-0292, Ibaraki, Japan

    Tetsuya Yamada, Yasumasa Takatsu & Masakazu Kasumi

  2. National Institute of Floricultural Science, 305-8519, Ibaraki, Japan

    Tetsuya Yamada & Kazuo Ichimura

  3. Wageningen University and Research Centre, P.O. Box 17, 6700 AA, Wageningen, The Netherlands

    Wouter G. van Doorn

Authors
  1. Tetsuya Yamada
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  2. Yasumasa Takatsu
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  3. Masakazu Kasumi
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  4. Kazuo Ichimura
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  5. Wouter G. van Doorn
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Correspondence to Tetsuya Yamada.

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Yamada, T., Takatsu, Y., Kasumi, M. et al. Nuclear fragmentation and DNA degradation during programmed cell death in petals of morning glory (Ipomoea nil). Planta 224, 1279–1290 (2006). https://doi.org/10.1007/s00425-006-0307-z

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  • DOI: https://doi.org/10.1007/s00425-006-0307-z

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