Growing Arabidopsis In Vitro: Cell Suspensions, In Vitro Culture, and Regeneration

  • Bronwyn J. Barkla
  • Rosario Vera-Estrella
  • Omar Pantoja
Part of the Methods in Molecular Biology book series (MIMB, volume 1062)


An understanding of basic methods in Arabidopsis tissue culture is beneficial for any laboratory working on this model plant. Tissue culture refers to the aseptic growth of cells, organs, or plants in a controlled environment, in which physical, nutrient, and hormonal conditions can all be easily manipulated and monitored. The methodology facilitates the production of a large number of plants that are genetically identical over a relatively short growth period. Techniques, including callus production, cell suspension cultures, and plant regeneration, are all indispensable tools for the study of cellular biochemical and molecular processes. Plant regeneration is a key technology for successful stable plant transformation, while cell suspension cultures can be exploited for metabolite profiling and mining. In this chapter we report methods for the successful and highly efficient in vitro regeneration of plants and production of stable cell suspension lines from leaf explants of both Arabidopsis thaliana and Arabidopsis halleri.

Key words

Callus Cell suspensions Plant regeneration Tissue culture Arabidopsis Organ regeneration 



Work in the authors’ lab is funded by DGAPA IN203913 to B.J.B., DGAPA 203711 to R.V.-E., and DGAPA 203112 and CONACyT IN79191 to O.P.


  1. 1.
    Krikorian AD, Berquam DL (1969) Plant cell and tissue cultures: the role of Haberlandt. Bot Rev 35:59–67CrossRefGoogle Scholar
  2. 2.
    Gautheret RJ (1937) Nouvelles recherches sur la culture du tissu cambial Cr hebd. Seanc Acad Sci 205:572–574Google Scholar
  3. 3.
    Nobécourt P (1937) Culture en serie de tissus vegetaux sur milieu artificiel. Cr hebd. Seanc Acad Sci 20:521–523Google Scholar
  4. 4.
    White PR (1939) Potentially unlimited growth of excised plant callus in an artificial medium. Am J Bot 26:59–64CrossRefGoogle Scholar
  5. 5.
    Steward FC (1958) Growth and organized development of cultured cells. II. Organization in cultures grown from freely suspended cells. Am J Bot 45:705–708CrossRefGoogle Scholar
  6. 6.
    Brown JT, Charlwood BV (1990) Organogenesis in callus culture. Methods Mol Biol 6:65–70PubMedGoogle Scholar
  7. 7.
    Loewenberg JR (1965) Callus cultures of Arabidopsis. Arabidopsis Inf Serv 2:34Google Scholar
  8. 8.
    Negrutiu I, Beeftink F, Jacobs M (1975) Arabidopsis thaliana as a model system in somatic cell genetics I. Cell and tissue culture. Plant Sci Lett 5:293–304CrossRefGoogle Scholar
  9. 9.
    Negrutiu I, Jacobs M (1975) Arabidopsis thaliana as a model system in somatic cell genetics II. Cell suspension culture. Plant Sci Lett 8:7–15Google Scholar
  10. 10.
    Lloyd AM, Barnason AR, Rogers SG, Byrne MC, Fraley RT, Horsch RB (1986) Transformation of Arabidopsis thaliana with Agrobacterium tumefaciens. Science 234:464–466PubMedCrossRefGoogle Scholar
  11. 11.
    Leonelli S (2007) Arabidopsis, the botanical Drosophila: from mouse cress to model organism. Endeavour 31:34–38PubMedCrossRefGoogle Scholar
  12. 12.
    Gamborg O, Miller R, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:151–158PubMedCrossRefGoogle Scholar
  13. 13.
    Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–479CrossRefGoogle Scholar
  14. 14.
    Hoagland DR, Arnon DI (1938) The water culture method for growing plants without soil. Calif Agric Exp Station Circ 347:1–39Google Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Bronwyn J. Barkla
    • 1
  • Rosario Vera-Estrella
    • 1
  • Omar Pantoja
    • 1
  1. 1.Instituto de BiotecnologíaUniversidad Nacional Autónoma de MéxicoCuernavacaMexico

Personalised recommendations