Skip to main content
SpringerLink
Log in

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

  • Original Article
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Abràmoff MD, Magalhães PJ, Ram SJ (2004) Image processing with ImageJ. Biophotonics Int 11:36–42

    Google Scholar 

  • Bassham DC (2007) Plant autophagy—more than a starvation response. Curr Opinion Plant Biol 10:587–593

    Article  CAS  Google Scholar 

  • Bassham DC (2015) Methods for analysis of autophagy in plants. Methods 75:181–188

    Article  CAS  PubMed  Google Scholar 

  • Blommaart EFC, Kraus U, Schellens JPM (1997) The phosphatidylinositol 3-kinase inhibitors wortmannin and LY294002 inhibit autophagy in isolated rat hepatocytes. Eur J Biochem 243:240–246

    Article  CAS  PubMed  Google Scholar 

  • De Verna JW, Myers JR, Collins GB (1987) Bypassing prefertilization barriers to hybridization in Nicotiana using in vitro pollination and fertilization. Theor Appl Genet 73:665–671

    Google Scholar 

  • Goldraij A, Roldán JA, Rojas HJ (2012) Early F-actin disorganization may be signaling vacuole disruption in incompatible pollen tubes of Nicotiana alata. Plant Signal Behav 7:1695–1697

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hatsugai N, Kuroyanagi M, Yamada K, Meshi T, Tsuda S, Kondo M et al (2004) A plant vacuolar protease, VPE, mediates virus-induced hypersensitive cell death. Science 305:855–858

    Article  CAS  PubMed  Google Scholar 

  • Hatsugai N, Yamada K, Goto-Yamada S, Hara-Nishimura I (2015) Vacuolar processing enzyme in plant programmed cell death. Front Plant Sci 6:234

    Article  PubMed  PubMed Central  Google Scholar 

  • Hofius D, Munch D, Bressendorff S, Mundy J, Petersen M (2011) Role of autophagy in disease resistance and hypersensitive response-associated cell death. Cell Death Differ 18:1257–1262

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kariya K, Demiral T, Sasaki T, Tsuchiya Y, Turkan I, Sano T et al (2013) A novel mechanism of aluminium-induced cell death involving vacuolar processing enzyme and vacuolar collapse in tobacco cell line BY-2. J Inorg Biochem 128:196–201

    Article  CAS  PubMed  Google Scholar 

  • Kobori S, Marubashi W (2004) Programmed cell death detected in interspecific hybrids of Nicotiana repanda x N. tomentosiformis expressing hybrid lethality. Breed Sci 54:347–350

    Article  Google Scholar 

  • Kostoff D (1930) Ontogeny, genetics, and cytology of Nicotiana hybrids. Genetica 12:33–139

    Article  Google Scholar 

  • Manabe T (1989) Temperature-dependent conditional lethality in interspecific hybrids between Nicotiana suaveolens Lehm. and N. tabacum L. In: Proceedings of the 6th International Congress of SABRAO, p 459–462

  • Marty F (1999) Plant vacuoles. Plant Cell 11:587–599

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Marubashi W, Kobayashi M (2002) Temperature-dependent apoptosis detected in hybrids between Nicotiana debneyi and N. tabacum expressing lethality. Plant Biotech 19:267–270

    Article  CAS  Google Scholar 

  • Marubashi W, Yamada T, Niwa M (1999) Apoptosis detected in hybrids between Nicotiana glutinosa and N. repanda expressing lethality. Planta 210:168–171

    Article  CAS  PubMed  Google Scholar 

  • Masuda Y, Yamada T, Marubashi W (2003) Time course analysis of apoptotic cell death during expression of hybrid lethality in hybrid tobacco cells (Nicotiana suaveolens × N. tabacum). Plant Cell Physiol 44:420–427

    Article  CAS  PubMed  Google Scholar 

  • Masuda Y, Yamada T, Kuboyama T, Marubashi W (2007) Identification and characterization of genes involved in hybrid lethality in hybrid tobacco cells (Nicotiana suaveolens × N. tabacum) using suppression subtractive hybridization. Plant Cell Rep 26:1595–1604

    Article  CAS  PubMed  Google Scholar 

  • Merkulova EA, Guiboileau A, Naya L, Masclaux-Daubresse C, Yoshimoto K (2014) Assessment and optimization of autophagy monitoring methods in Arabidopsis roots indicate direct fusion of autophagosomes with vacuoles. Plant Cell Physiol 55:715–726

    Article  CAS  PubMed  Google Scholar 

  • Mino M, Maekawa K, Ogawa K, Yamagishi H, Inoue M (2002) Cell death processes during expression of hybrid lethality in interspecific F1 hybrid between Nicotiana gossei Domin and Nicotiana tabacum. Plant Physiol 130:1776–1787

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mino M, Misaka Y, Ueda J, Ogawa K, Inoue M (2005) Hybrid lethality of cultured cells of an interspecific F1 hybrid of Nicotiana gossei Domin and N. tabacum L. Plant Cell Rep 24:179–188

    Article  CAS  PubMed  Google Scholar 

  • Mino M, Murata N, Date S, Inoue M (2007) Cell death in seedlings of the interspecific hybrid of Nicotiana gossei and N. tabacum; possible role of knob-like bodies formed on tonoplast in vacuolar-collapse-mediated cell death. Plant Cell Rep 26:407–419

    Article  CAS  PubMed  Google Scholar 

  • Obara K, Kuriyama H, Fukuda H (2001) Direct evidence of active and rapid nuclear degradation triggered by vacuole rupture during programmed cell death in Zinnia. Plant Physiol 125:615–626

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Palta JP, Li PH (1978) Cell membrane properties in relation to freezing injury. In: Li PH, Sakai A (eds) Plant cold hardiness and freezing stress. Mechanisms and crop implications. Academic Press, New York, pp 93–115

    Chapter  Google Scholar 

  • Phillips LL, Reid RK (1975) Interspecific incompatibility in Gossypium. II. Light and electron microscopic studies of cell necrosis and tumorigenesis in hybrids of G. klotzschianum. Am J Bot 62:790–796

    Article  Google Scholar 

  • Shinozaki Y, Tanaka T, Ogiwara I, Kanekatsu M, van Doorn WG, Yamada T (2014) Expression of an AtNAP gene homolog in senescing morning glory (Ipomoea nil) petals of two cultivars with a different flower life span. J Plant Physiol 171:633–638

    Article  CAS  PubMed  Google Scholar 

  • Stebbins GL (1966) Reproductive isolation and the origin of species. In: Processes of organic evolution. Prentice-Hall, New Jersey, p 85–112

  • Strober W (2001) Trypan blue exclusion test of cell viability. Curr Protoc Immunol 111:A–3B

    Google Scholar 

  • Sukumaran NP, Weiser CJ (1972) An excised leaflet test for evaluating potato frost tolerance. HortScience 7:467–468

    Google Scholar 

  • Takatsuka C, Inoue Y, Matsuoka K, Moriyasu Y (2004) 3-Methyladenine inhibits autophagy in tobacco culture cells under sucrose starvation conditions. Plant Cell Physiol 45:265–274

    Article  CAS  PubMed  Google Scholar 

  • Tezuka T, Kuboyama T, Matsuda T, Marubashi W (2010) Seven of eight species in Nicotiana section Suaveolentes have common factors leading to hybrid lethality in crosses with Nicotiana tabacum. Ann Bot 106:267–276

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • van Doorn WG (2005) Plant programmed cell death and the point of no return. Trends Plant Sci 10:478–483

    Article  CAS  PubMed  Google Scholar 

  • van Doorn WG (2011) Classes of programmed cell death in plants, compared to those in animals. J Exp Bot 62:4749–4761

    Article  CAS  PubMed  Google Scholar 

  • van Doorn WG, Beers EP, Dangl JL, Franklin-Tong VE, Gallois P, Hara-Nishimura I et al (2011) Morphological classification of plant cell deaths. Cell Death Differ 18:1241–1246

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Watanabe H, Marubashi W (2004) Temperature-dependent programmed cell death detected in hybrids between Nicotiana langsdorffii and N. tabacum expressing lethality. Plant Biotech 21:151–154

    Article  CAS  Google Scholar 

  • Wilkins KA, Bancroft J, Bosch M, Ings J, Smirnoff N, Franklin-Tong VE (2011) Reactive oxygen species and nitric oxide mediate actin reorganization and programmed cell death in the self-incompatibility response of papaver. Plant Physiol 156:404–416

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wu YT, Tan HL, Shui G, Bauvy C, Huang Q, Wenk MR et al (2010) Dual role of 3-methyladenine in modulation of autophagy via different temporal patterns of inhibition on class I and III phosphoinositide 3-kinase. J Biol Chem 285:10850–10861

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yamada T, Marubashi W, Niwa M (1999) Detection of four lethality types in interspecific crosses among Nicotiana species through the use of three rescue methods for lethality. Breed Sci 49:203–210

    Article  Google Scholar 

  • Yamada T, Marubashi W, Niwa M (2000) Apoptotic cell death induces temperature-sensitive lethality in hybrid seedlings and calli derived from the cross of Nicotiana suaveolens × N. tabacum. Planta 211:614–622

    Article  CAS  PubMed  Google Scholar 

  • Yamada T, Marubashi W, Niwa M (2001) Facile induction of apoptosis into plant cells associated with temperature-sensitive lethality shown on interspecific hybrid from the cross Nicotiana suaveolens × N. tabacum. Plant Cell Physiol 42:204–213

    Article  CAS  PubMed  Google Scholar 

  • Zeven AC (1981) Eighth supplementary list of wheat varieties classified according to their genotypes for hybrid necrosis. Euphytica 30:521–539

    Article  Google Scholar 

Download references

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).

Author information

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
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Saori Nihei
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Naoto Miyakawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tadashi Hirasawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Motoki Kanekatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Wataru Marubashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Wouter G. van Doorn
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tetsuya Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tetsuya Yamada.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Communicated by F. Sato.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

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

Keywords

Search

Navigation