Journal of Plant Research

, Volume 126, Issue 2, pp 243–250 | Cite as

Acquisition of embryogenic competency does not require cell division in carrot somatic cell

  • Akira KikuchiEmail author
  • Masashi Asahina
  • Motoki Tanaka
  • Shinobu Satoh
  • Hiroshi Kamada
Regular Paper


Totipotency is the ability of a cell to regenerate the entire organism, even after previous differentiation as a specific cell. When totipotency is coupled with active cell division, it was presumed that cell division is essential for this expression. Here, using the stress-induction system of somatic embryos in carrots, we show that cell division is not essential for the expression of totipotency in somatic/embryonic conversion. Morphological and histochemical analyses showed that the cell did not divide during embryo induction. Inhibitors of cell division did not affect the rate of somatic embryo formation. Our results indicate that the newly acquired trait of differentiation appears without cell division, but does not arise with cell division as a newborn cell.


Carrot (Daucus carotaCell division Somatic embryogenesis Totipotency 









Dimethyl sulfoxide







This work was supported in part by the Grants-in-Aid program from the Ministry of Education, Culture, Sports, Science and Technology of Japan (Grant no. 23570045) and by the joint research program “Plant Transgenic Research Design, University of Tsukuba.”


  1. Asahina M, Iwai H, Kikuchi A, Yamaguchi S, Kamiya Y, Kamada H, Satoh S (2002) Gibberellin produced in the cotyledon is required for cell division during tissue reunion in the cortex of cut cucumber and tomato hypocotyls. Plant Physiol 129:201–210PubMedCrossRefGoogle Scholar
  2. Evans MJ, Kaufman MH (1981) Establishment in culture of pluripotential cells from mouse embryos. Nature 292:154–156PubMedCrossRefGoogle Scholar
  3. Fukuda H, Komamine A (1980) Establishment of an experimental system for the study of tracheary element differentiation from single cells isolated from the mesophyll of Zinnia elegans. Plant Physiol 65:57–60PubMedCrossRefGoogle Scholar
  4. Fukuda H, Komamine A (1981) Relationship between tracheary element differentiation and the cell cycle in single cells isolated from the mesophyll of Zinnia elegans. Physiol Plant 52:423–430CrossRefGoogle Scholar
  5. Gao S, Chung YG, Parseghian MH, King GJ, Adashi EY, Latham KE (2004) Rapid H1 linker histone transitions following fertilization or somatic cell nuclear transfer, evidence for a uniform developmental program in mice. Dev Biol 266:62–75PubMedCrossRefGoogle Scholar
  6. Ikeda-Iwai M, Umehara M, Satoh S, Kamada H (2003) Stress-induced somatic embryogenesis in vegetative tissues of Arabidopsis thaliana. Plant J 34:107–114PubMedCrossRefGoogle Scholar
  7. Illmensee K, Mintz B (1976) Totipotency and normal differentiation of single teratocarcinoma cells cloned by injection into blastocysts. Proc Natl Acad Sci USA 73:549–553PubMedCrossRefGoogle Scholar
  8. Kamada H, Harada H (1979) Studies on organogenesis in carrot tissue culture. I. Effects of growth regulation on somatic embryogenesis and root formation. Z Pflanzenphysiol 91:225–266Google Scholar
  9. Kamada H, Kobayashi K, Kiyosue T, Harada H (1989) Stress-induced somatic embryogenesis in carrot and its application to synthetic seed production. In Vitro Cell Dev Biol 25:1163–1166CrossRefGoogle Scholar
  10. Kamada H, Tachikawa Y, Saitou T, Harada H (1994) Heat stress induction of carrot somatic embryogenesis. Plant Tissue Cult Lett 11:229–232CrossRefGoogle Scholar
  11. Kates JR, Jones RF (1964) The control of gametic differentiation in liquid cultures of Chlamydomonas. J Cell Comp Physiol 63:157–164CrossRefGoogle Scholar
  12. Kikuchi A, Sanuki N, Higashi K, Koshiba T, Kamada H (2006) Abscisic acid and stress treatment are essential for the acquisition of embryogenic competence by carrot somatic cells. Planta 223:637–645PubMedCrossRefGoogle Scholar
  13. Kiyosue T, Kamada H, Harada H (1989) Induction of somatic embryogenesis by salt stress in carrot. Plant Tissue Cult Lett 6:162–164CrossRefGoogle Scholar
  14. Kiyosue T, Takano K, Kamada H, Harada H (1990) Induction of somatic embryogenesis in carrot by heavy metal ions. Can J Bot 68:2021–2033CrossRefGoogle Scholar
  15. Kiyosue T, Yamaguchi-Shinozaki K, Shinozaki K, Kamada H, Harada H (1993) cDNA cloning of ECP40, an embryogenic-cell protein in carrot, and its expression during somatic and zygotic embryogenesis. Plant Mol Biol 21:1053–1068PubMedCrossRefGoogle Scholar
  16. Kumlehn J, Loerz H (1999) Monitoring sporophytic development of individual microspores of barley (Hordeum vulgare L.). In: Clement C, Paccini E, Audran JC (eds) Anther and pollen, from biology to biotechnology. Springer, Berlin, pp 183–190CrossRefGoogle Scholar
  17. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  18. Nagata T, Nemoto Y, Hasezawa S (1992) Tobacco BY-2 cell line as the “HeLa” cell in the cell biology of higher plants. Int Rev Cytol 132:1–30CrossRefGoogle Scholar
  19. Nomura K, Komamine A (1985) Identification and isolation of single cells that produce somatic embryos at a high frequency in a carrot suspension culture. Plant Physiol 79:988–991PubMedCrossRefGoogle Scholar
  20. Reinert J (1958) Morphogenese und ihre Kontrolle an Gewebekulturen aus Carotten. Naturwissenschaften 45:344–345CrossRefGoogle Scholar
  21. Samuels AL, Meehl J, Lipe M, Staehelin LA (1998) Optimizing conditions for tobacco BY-2 cell cycle synchronization. Protoplasma 202:232–236CrossRefGoogle Scholar
  22. Santos F, Dean W (2004) Epigenetic reprogramming during early development in mammals. Reproduction 127:643–651PubMedCrossRefGoogle Scholar
  23. Santos TA, Dias C, Henriques P, Brito R, Barbosa A, Regateiro F, Santos AA (2003) Cytogenetic analysis of spontaneously activated noninseminated oocytes and parthenogenetically activated failed fertilized human oocytes—implications for the use of primate parthenotes for stem cell production. J Assist Reprod Genet 20:122–130PubMedCrossRefGoogle Scholar
  24. Shibukawa T, Yazawa K, Kikuchi A, Kamada H (2009) Possible involvement of DNA methylation on expression regulation of carrot LEC1 gene in its 5′-upstream region. Gene 437:22–31PubMedCrossRefGoogle Scholar
  25. Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissue cultured in vitro. Symp Soc Exp Biol 11:118–130PubMedGoogle Scholar
  26. Steward FC, Mapes MO, Smith J (1958) Growth and organized development of cultured cells. I. Growth and division of freely suspended cells. Am J Bot 45:707–708Google Scholar
  27. Suzuki T, Sasaki N, Sakai A, Kawano S, Kuroiwa T (1995) Localization of organelle DNA synthesis within the root apical meristem of rice. J Exp Bot 46:19–25CrossRefGoogle Scholar
  28. Tanaka M, Kikuchi A, Kamada H (2008) The Arabidopsis histone deacetylases HDA6 and HDA19 contribute to the repression of embryonic properties after germination. Plant Physiol 146:149–161PubMedCrossRefGoogle Scholar
  29. Tanaka M, Kikuchi A, Kamada H (2009) Isolation of putative embryo-specific genes using stress induction of carrot somatic embryos. Breed Sci 59:37–46CrossRefGoogle Scholar
  30. Teranishi T, Tanaka M, Kimoto S, Ono Y, Miyakoshi K, Kono T, Yoshimura Y (2004) Rapid replacement of somatic linker histones with the oocyte-specific linker histone H1foo in nuclear transfer. Dev Biol 266:76–86PubMedCrossRefGoogle Scholar
  31. Wakayama T, Perry AC, Zuccotti M, Johnson KR, Yanagimachi R (1998) Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394:369–374PubMedCrossRefGoogle Scholar
  32. Wilde HD, Nelson WS, Booij H, De Vries SC, Thomas TL (1988) Gene-expression programs in embryogenic and non-embryogenic carrot cultures. Planta 176:205–211CrossRefGoogle Scholar
  33. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KH (1997) Viable offspring derived from fetal and adult mammalian cells. Nature 385:810–813 (erratum in Nature 386:200)Google Scholar
  34. Yazawa K, Takahata K, Kamada H (2004) Isolation of the gene encoding carrot leafy cotyledon1 and expression analysis during somatic zygotic embryogenesis. Plant Physiol Biochem 42:215–223PubMedCrossRefGoogle Scholar
  35. Zhang L, Qiu Z, Hu Y, Yang F, Yan S, Zhao L, Li B, He S, Huang M, Li J, Li L (2011) ABA treatment of germinating maize seeds induces VP1 gene expression and selective promoter-associated histone acetylation. Physiol Plant 143:287–296PubMedCrossRefGoogle Scholar

Copyright information

© The Botanical Society of Japan and Springer 2012

Authors and Affiliations

  • Akira Kikuchi
    • 1
    Email author
  • Masashi Asahina
    • 1
    • 2
  • Motoki Tanaka
    • 1
    • 3
  • Shinobu Satoh
    • 1
  • Hiroshi Kamada
    • 1
  1. 1.Faculty of Life and Environmental Sciences, Gene Research CenterUniversity of TsukubaTsukubaJapan
  2. 2.Department of BiosciencesTeikyo UniversityUtsunomiyaJapan
  3. 3.SDS Biotech K.K., Tsukuba Research and Technology CenterTsukubaJapan

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