In Vitro Assay for Induction of Adventitious Rooting on Intact Arabidopsis Hypocotyls

  • Hoang Khai Trinh
  • Inge Verstraeten
  • Danny Geelen
Part of the Methods in Molecular Biology book series (MIMB, volume 1761)


Adventitious roots (AR) are de novo formed roots that emerge from any part of the plant or from callus in tissue culture, except root tissue. The plant tissue origin and the method by which they are induced determine the physiological properties of emerged ARs. Hence, a standard method encompassing all types of AR does not exist. Here we describe a method for the induction and analysis of AR that emerge from the etiolated hypocotyl of dicot plants. The hypocotyl is formed during embryogenesis and shows a determined developmental pattern which usually does not involve AR formation. However, the hypocotyl shows propensity to form de novo roots under specific circumstances such as removal of the root system, high humidity or flooding, or during de-etiolation. The hypocotyl AR emerge from a pericycle-like cell layer surrounding the vascular tissue of the central cylinder, which is reminiscent to the developmental program of lateral roots. Here we propose an easy protocol for in vitro hypocotyl AR induction from etiolated Arabidopsis seedlings.

Key words

Etiolated seedlings Hypocotyls Adventitious roots 


  1. 1.
    Bellini C, Pacurar DI, Perrone I (2014) Adventitious roots and lateral roots: similarities and differences. Annu Rev Plant Biol 65:639–666CrossRefPubMedGoogle Scholar
  2. 2.
    Verstraeten I, Schotte S, Geelen D (2014) Hypocotyl adventitious root organogenesis differs from lateral root development. Front Plant Sci 5(9):1–13Google Scholar
  3. 3.
    da Rocha Correa L, Troleis J, Mastroberti AA, Mariath JEA, Fett-Neto AG (2012) Distinct modes of adventitious rooting in Arabidopsis thaliana. Plant Biol 14(1):100–109Google Scholar
  4. 4.
    Sorin C et al (2005) Auxin and light control of adventitious rooting in Arabidopsis require ARGONAUTE1. Plant Cell 17:1343–1359CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    de Klerk G-J, van der Krieken W, de Jong JC (2012) The formation of adventitious roots: new concepts , new possibilities. In Vitro Cell Dev Biol Plant 35(3):189–199CrossRefGoogle Scholar
  6. 6.
    Verstraeten I, Beeckman T, Geelen D (2013) Adventitious root induction in arabidopsis thaliana as a model for in vitro root organogenesis. Methods Mol Biol 959:159–175CrossRefPubMedGoogle Scholar
  7. 7.
    Schindelin J et al (2013) Fiji - an open source platform for biological image analysis. Nat Methods 9(7):676–682CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  • Hoang Khai Trinh
    • 1
  • Inge Verstraeten
    • 2
  • Danny Geelen
    • 1
  1. 1.In Vitro Biology and Horticulture, Department of Plant Production, Faculty of Bioscience EngineeringUniversity of GhentGhentBelgium
  2. 2.Institute of Science and Technology Austria (IST Austria)KlosterneuburgAustria

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