Journal Article

Facile Induction of Apoptosis into Plant Cells Associated with Temperature-Sensitive Lethality Shown on Interspecific Hybrid from the Cross <i>Nicotiana suaveolens</i> × <i>N. tabacum</i>

Tetsuya Yamada, Wataru Marubashi and Masaru Niwa

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 2, pages 204-213
Published in print February 2001 | ISSN: 0032-0781
Published online February 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce029
Facile Induction of Apoptosis into Plant Cells Associated with Temperature-Sensitive Lethality Shown on Interspecific Hybrid from the Cross Nicotiana suaveolens × N. tabacum

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Two lines of suspension culture cells were obtained from a hybrid seedling of Nicotiana suaveolens Lehm. × N. tabacum L. cv. Hicks-2 expressing temperature-sensitive lethality. One of them (LH line) was inducible cell death in accordance with the lethality at 28°C but not under high-temperature conditions (36°C). Another one (SH line) lost the lethality and survived at 28°C. The cells of LH line showed apoptotic changes when they were cultured at 28°C. Fragmentation of nuclei was correlated with the lethality in the cells, as confirmed by fluorimetry of the nuclear DNA using laser scanning cytometry. Agarose gel analysis of DNA extracted from the cells expressing the lethality revealed a specific ladder pattern suggesting nucleosomal fragmentation that is one of the biochemical characteristics of apoptosis. From these facts, we confirmed that the process of cell death leading to hybrid lethality in the cells is certainly apoptosis. Hybrid cells were used in the experiments to estimate the point of no return in temperature-sensitive lethality and to examine the influence of cation in DNA fragmentation during apoptosis. The utility of hybrid cells as an experimental system for studies of hybrid lethality and apoptosis in plants was confirmed.

Keywords: Key words: Apoptosis — DNA fragmentation — Interspecific hybrid — Suspension culture cell — Temperature-sensitive lethality — Tobacco.; Abbreviations: BAP, 6-benzylaminopurine; CTAB, cetyltrimethylammonium bromide; DAPI, 4′-6-diamino-2-phenylindole dihydrochloride; FDA, fluorescein diacetate; NAA, α-naphthalenacetic acid; PI, propidium iodide; TUNEL, terminal deoxyribonucleotidyl transferase-mediated dUTP nick-end labeling.

Journal Article.  6805 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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