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Identification and characterization of genes involved in hybrid lethality in hybrid tobacco cells (Nicotiana suaveolens × N. tabacum) using suppression subtractive hybridization

Plant Cell Reports volume 26, pages 1595–1604 (2007)Cite this article

Abstract

Hybrid lethality is an important problem for cross-breeding; however, its molecular mechanism is not clear. The purpose of the present study was to identify the genes expressed during hybrid lethality in the hybrid cells (Nicotiana suaveolens × N. tabacum). In order to identify these genes, we employed suppression subtractive hybridization (SSH) between RNA isolated from cells expressing lethality (lethal hybrid line; LH line) and cells overcoming lethality fortuitously (a surviving hybrid line; SH line). Four populations of cDNA were created from the time points corresponding to before and during induction, and at and after the point of no return (PNR) during the process of programmed cell death (PCD) that occurs during hybrid lethality. By SSH and following dot-blot macroarray analysis, 99 genes out of 138 isolated clones were identified as hybrid lethality-related (HLR) genes. Quantitative real-time PCR analysis data indicated that ten clones were expressed specifically in LH line cells. The HLR genes in these clones show homology to genes involved in disease resistance, ethylene-induced reactions, phosphorylation, ubiquitination, jasmonic acid-related reactions, calcium signaling and self-incompatibility. These data suggested that at least some parts of the mechanism of hybrid lethality are shared with those of the putative functions of the HLR gene-related pathways.

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Fig. 1

Abbreviations

HLR:

Hybrid lethality related

LH:

Lethal hybrid

PCD:

Programmed cell death

PNR:

Point of no-return

SH:

Surviving hybrid

SSH:

Suppression subtractive hybridization

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Acknowledgments

This work was partly supported by the Sasakawa Scientific Research Grant from The Japan Science Society, and a Grant-in-Aid for Scientific Research (C) No. 15580003 from the Ministry of Education, Culture, Sports, Science and Technology, Japan.

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Authors and Affiliations

  1. Laboratory of Plant Chromosome and Gene Stock, Graduate School of Science, Hiroshima University, 1-4-3 Kagamiyama, Higashi-Hiroshima-city, Hiroshima, 739-8526, Japan

    Yu Masuda

  2. Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Tokyo, 183-8509, Japan

    Tetsuya Yamada

  3. School of Agriculture, Ibaraki University, Ami, Ibaraki, 300-0393, Japan

    Tsutomu Kuboyama

  4. School of Agriculture, Meiji University, 1-1-1 Higashimita, Kawasaki, Kanagawa, 214-8571, Japan

    Wataru Marubashi

Authors
  1. Yu Masuda
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  2. Tetsuya Yamada
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  3. Tsutomu Kuboyama
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  4. Wataru Marubashi
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Corresponding author

Correspondence to Wataru Marubashi.

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Communicated by C.F. Quiros.

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Masuda, Y., Yamada, T., Kuboyama, T. et al. 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 (2007). https://doi.org/10.1007/s00299-007-0352-5

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  • DOI: https://doi.org/10.1007/s00299-007-0352-5

Keywords

  • Hybrid lethality
  • Nicotiana suaveolens
  • N. tabacum
  • Programmed cell death
  • Suppression subtractive hybridization