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Time course of programmed cell death, which included autophagic features, in hybrid tobacco cells expressing hybrid lethality

Plant Cell Reports volume 35, pages 2475–2488 (2016)Cite this article

Abstract

Key message

PCD with features of vacuolar cell death including autophagy-related features were detected in hybrid tobacco cells, and detailed time course of features of vacuolar cell death were established.

Abstract

A type of interspecific Nicotiana hybrid, Nicotiana suaveolens × N. tabacum exhibits temperature-sensitive lethality. This lethality results from programmed cell death (PCD) in hybrid seedlings, but this PCD occurs only in seedlings and suspension-cultured cells grown at 28 °C, not those grown at 36 °C. Plant PCD can be classified as vacuolar cell death or necrotic cell death. Induction of autophagy, vacuolar membrane collapse and actin disorganization are each known features of vacuolar cell death, but observed cases of PCD showing all these features simultaneously are rare. In this study, these features of vacuolar cell death were evident in hybrid tobacco cells expressing hybrid lethality. Ion leakage, plasma membrane disruption, increased activity of vacuolar processing enzyme, vacuolar membrane collapse, and formation of punctate F-actin foci were each evident in these cells. Transmission electron microscopy revealed that macroautophagic structures formed and tonoplasts ruptured in these cells. The number of cells that contained monodansylcadaverine (MDC)-stained structures and the abundance of nine autophagy-related gene transcripts increased just before cell death at 28 °C; these features were not evident at 36 °C. We assessed whether an autophagic inhibitor, wortmannin (WM), influenced lethality in hybrid cells. After the hybrid cell began to die, WM suppressed increases in ion leakage and cell deaths, and it decreased the number of cells containing MDC-stained structures. These results showed that several features indicative of autophagy and vacuolar cell death were evident in the hybrid tobacco cells subject to lethality. In addition, we documented a detailed time course of these vacuolar cell death features.

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Acknowledgments

This work was performed under the Global Innovation Research Organization at the Tokyo University of Agriculture and Technology and was partly supported by a Grant-in Aid for Challenging Exploratory Research (No. 246580004) to T. Yamada from the Japan Society for the Promotion of Science (JSPS).

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Authors and Affiliations

  1. United Graduate School of Agricultural Science, Tokyo University of Agriculture and Technology, Tokyo, Japan

    Naoya Ueno, Tadashi Hirasawa, Motoki Kanekatsu & Tetsuya Yamada

  2. Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo, Japan

    Saori Nihei, Naoto Miyakawa, Tadashi Hirasawa, Motoki Kanekatsu & Tetsuya Yamada

  3. Faculty of Agricultural Science, Meiji University, Kanagawa, Japan

    Wataru Marubashi

  4. Mann Laboratory, Department of Plant Sciences, University of California, Davis, CA, USA

    Wouter G. van Doorn

  5. Global Innovation Research Organization, Tokyo University of Agriculture and Technology, Tokyo, Japan

    Tadashi Hirasawa, Wouter G. van Doorn & Tetsuya Yamada

Authors
  1. Naoya Ueno
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  2. Saori Nihei
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  3. Naoto Miyakawa
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  4. Tadashi Hirasawa
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  5. Motoki Kanekatsu
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  6. Wataru Marubashi
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  7. Wouter G. van Doorn
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  8. Tetsuya Yamada
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Correspondence to Tetsuya Yamada.

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Communicated by F. Sato.

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Ueno, N., Nihei, S., Miyakawa, N. et al. Time course of programmed cell death, which included autophagic features, in hybrid tobacco cells expressing hybrid lethality. Plant Cell Rep 35, 2475–2488 (2016). https://doi.org/10.1007/s00299-016-2048-1

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  • DOI: https://doi.org/10.1007/s00299-016-2048-1

Keywords

  • Autophagy
  • Hybrid lethality
  • Tobacco
  • Programmed cell death
  • Vacuolar cell death