Plant Cell Reports

, Volume 23, Issue 6, pp 371–376 | Cite as

Efficient plant regeneration from suspension cells of Allium cepa L.

  • Wenbo Zhang
  • Xiaofei Lin
  • Hiroyoshi Takano
  • Susumu Takio
  • Kanji Ono
Cell Biology and Morphogenesis


Plant regeneration from calli of three cultivars of Allium cepa (Senshuki, O·Pki and Shojovaka) was investigated. Callus was induced on four variations of BDS medium containing different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BAP). The regeneration frequency of calli of cvs. Senshuki and O·Pki subcultured on solid MS medium supplemented with BAP ranged from 50% to 80%; this frequency decreased to less than 30% after subculture in the dark in liquid BDS medium. By repeating the dark/light transitions of the culture protocol and by selecting for green cell clusters, we were able to increase the regeneration frequency to more than 80% in all three cultivars. These cell clusters maintained a high regeneration capacity in subsequent subcultures in the absence of light for 2 months. Most (97%) of the regenerated plantlets had a normal diploid karyotype (2n=16) that was identical to that of the mother plants, although 3% of the regenerated plants of cv. Shojovaka had a tetraploid karyotype.


Allium cepaMonocotyledon Plant regeneration Ploidy Tissue culture 





2,4-Dichlorophenoxyacetic acid



This investigation was supported by a grant from the Program for Promotion of Basic Research Activities for Innovative Biosciences (PROBRAIN; to H.T.).


  1. Dunstan DI, Short KC (1977) Improved growth of tissue cultures of the onion, Allium cepa. Physiol Plant 41:70–72Google Scholar
  2. Dunstan DI, Short KC (1978) Shoot production from onion callus tissue cultures. Sci Hortic 9:99–110CrossRefGoogle Scholar
  3. Eady CC, Butler RC, Suo Y (1998) Somatic embryogenesis and plant regeneration from immature embryo cultures of onion (Allium.cepa L.). Plant Cell Rep 18:111–116CrossRefGoogle Scholar
  4. Eady CC, Weld RJ, Lister CE (2000) Agrobacterium tumefaciens-mediated transformation and transgenic-plant regeneration of onion (Allium cepa L.). Plant Cell Rep 19:376–381CrossRefGoogle Scholar
  5. Eady CC, Davis S, Farrant J, Reader J, Kenel F (2003a) Agrobacterium tumefaciens-mediated transformation and regeneration of herbicide-resistant onion (Allium cepa L.) plants. Ann Appl Biol 142:213–217Google Scholar
  6. Eady CC, Reader J, Davis S, Dale T (2003b) Inheritance and expression of introduced DNA in transgenic onion plants (Allium cepa L.). Ann Appl Biol 142:219–224Google Scholar
  7. Gould AR (1979) Chromosomal and phenotypic stability during regeneration of whole plants from tissue cultures of Brachycome dichromosomatica (2n=4). Aust J Bot 27:117–121Google Scholar
  8. Hansen EE, Hubstenberger JF, Phillips GC (1995) Regeneration of shoots from cell suspension-derived protoplasts of Allium cepa. Plant Cell Rep 15:8–11Google Scholar
  9. Hussey G, Falavigna A (1980) Origin and production of in vitro adventitious shoots in the onion, Allium cepa L. J Exp Bot 31:1675–1686Google Scholar
  10. Karim MA, Adachi T (1997) Cell suspension, isolation and culture of protoplasts of Allium cepa. Plant Cell Tissue Organ Cult 51:43–47CrossRefGoogle Scholar
  11. Kim JW, Soh WY (1996) Plant regeneration through somatic embryogenesis from suspension cultures of Allium fistulosum L. Plant Sci 114:215–220CrossRefGoogle Scholar
  12. Lee K, Ono K (1999) Chromosomal variation in callus lines and regenerated plantlets from three cultivars of Allium fistulosum L. (2n=16). Cytologia 64:465–478Google Scholar
  13. Lee K, Ono K (2000) Somatic embryogenesis pathways in cultured Allium fistulosum cells. Cytologia 65:343–349Google Scholar
  14. Mitra J, Mapes MO, Steward FC (1960) Growth and organized development of cultured cells. IV. The behavior of the nucleus. Am J Bot 47:357–367Google Scholar
  15. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497Google Scholar
  16. Phillips GC, Luteyn KJ (1983) Effect of picloram and other auxins on onion tissue cultures. J Am Soc Hortic Sci 108:948–953Google Scholar
  17. Song P, Peffley EB (1994) Plant regeneration from suspension cultures of Allium fistulosum and A. fistulosum × A. cepa interspecific hybrid. Plant Sci 98:63–68CrossRefGoogle Scholar
  18. Van der Valk P, Scholten OE, Verstappen F, Jansen RC, Dons JJM (1992) High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species. Plant Cell Tissue Organ Cult 30:181–191Google Scholar
  19. Wang H, Debergh P (1995) Somatic embryogenesis and plant regeneration in garden leek. Plant Cell Tissue Organ Cult 43:21–28Google Scholar
  20. Zheng SJ, Henken B, Sofiari E, Keizer P, Jacobsen E, Kik C, Krens FA (1999) Effect of cytokinins and lines on plant regeneration from long-term callus and suspension cultures of Allium cepa L. Euphytica 108:83–90CrossRefGoogle Scholar
  21. Zheng SJ, Henken B, Sofiari E, Jacobsen E, Krens FA, Kik C (2001a) Molecular characterization of transgenic shallots (Allium cepa L.) by adaptor ligation PCR (AL-PCR) and sequencing of genomic DNA flanking T-DNA borders. Transgenic Res 10:237–245CrossRefPubMedGoogle Scholar
  22. Zheng SJ, Khrustaleva L, Henken B, Sofiari E, Jacobsen E, Kik C (2001b) Agrobacterium tumefaciens-mediated transformation of Allium cepa L.: the production of transgenic onions and shallots. Mol Breed 7:101–115CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Wenbo Zhang
    • 1
  • Xiaofei Lin
    • 1
  • Hiroyoshi Takano
    • 1
  • Susumu Takio
    • 2
  • Kanji Ono
    • 3
  1. 1.Graduate School of Science and TechnologyKumamoto UniversityKumamotoJapan
  2. 2.Center for Marine Environment StudiesKumamoto UniversityKumamotoJapan
  3. 3.Department of Biological Science, Faculty of ScienceKumamoto UniversityKumamotoJapan

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