Skip to main content

High-frequency direct shoot regeneration and continuous production of rapid-cycling Brassica oleracea in vitro


An in vitro method was developed for high-frequency shool regeneration and continuous production of rapid-cycling B. oleracea in large numbers. The high regenerative capacity was tissue-dependent. Developmental polarity (apical and basal ends) of the explants appeared to play a regulatory role in shoot morphogenesis in this system. High-frequency shoot regeneration was obtained with N6-benzyladenine or thidiazuron-supplemented media. Delayed and reduced regenerative ability of cultures in air-tight vessels and the dramatic suppression of shoot regeneration in internodal explants by the ethylene precursor 1-aminocyclopropane-1-carboxylic acid implicate a possible involvement of ethylene in shoot morphogenesis in this species. Rotting of regenerated shoots of B. oleracea occurred readily on α-naphthaleneacetic acid-supplemented media. Rooted plantlets were successfully established in soil and developed normal fertile flowers and viable seeds.

This is a preview of subscription content, access via your institution.


  • Arumuganathan, K.; Earle, E. D. Nuclear DNA content of some important plant species. Plant Mol. Biol. Rep. 9:208–218; 1991.

    CAS  Google Scholar 

  • Aslam, F. N.; MacDonald, M. V.; Ingram, D. S. Rapid-cycling Brassica species: anther culture potential of B. campestris L. and B. napus L. New Phytol. 115:1–9; 1990.

    Article  Google Scholar 

  • Bajaj, Y. P. S.; Nietsch, P. In vitro propagation of red cabbage (Brassica oleracea L. var capitata). J. Exp. Bot. 26:883–890; 1975.

    Article  CAS  Google Scholar 

  • Berthomieu, P.; Jouanin, L. Transformation of rapid cycling cabbage (Brassica oleracea var capitata) with Agrobacterium rhizogenes. Plant Cell Rep. 11:334–338; 1992.

    Article  CAS  Google Scholar 

  • Chi, G. L.; Pua, E. C.; Goh, C. J. Role of ethylene on de novo shoot regeneration from cotyledonary explants of Brassica campestris spp. pekinensis (Lour) Olsson in vitro. Plant Physiol. 96:178–183; 1991.

    PubMed  CAS  Google Scholar 

  • Fosket, D. E. Plant growth and development. California: Academic Press; 1994:363–369.

    Google Scholar 

  • Hansen, L. N.; Earle, E. D. Regeneration of plants from protoplasts of rapid cycling Brassica oleracea L. Plant Cell Rep. 13:335–339; 1994.

    Article  CAS  Google Scholar 

  • Kik, C.; Zaal, M. A. C. M. Protoplast regeneration from Brassica oleracea “rapid cycling’ and other varieties. Plant Cell Tiss. Organ Cult. 35:107–114; 1993.

    Article  Google Scholar 

  • Kumar, P. P.; Lakshmanan, P.; Thorpe, T. A. Regulation of morphogenesis in plant tissue culture by ethylene. In Vitro Cell Dev. Biol. Plant 34:94–103; 1998.

    CAS  Article  Google Scholar 

  • Lakshmanan, P.; Ng, S. K.; Loh, C. S.; Goh, C. J. Auxin, cytokinin and ethylene differentially regulate specific developmental states associated with shoot bud morphogenesis in leaf tissues of mangosteen (Garcinia mangostana L.) cultured in vitro. Plant Cell Physiol. 38:59–64; 1997.

    CAS  Google Scholar 

  • Larkin, P. J.; Scowcroft, W. R. Somaclonal variation—a novel source of variability from cell cultures for plant improvement. Theor. Appl. Genet. 60:197–214; 1981.

    Article  Google Scholar 

  • Leung, H.; Williams, P. Cytoplasmic male sterile Brassica campestris breeding lines with resistance to clubroot, turnip mosaic, and downy mildew. HortScience 18:774–775; 1983.

    Google Scholar 

  • Loudon, P. T.; Nelson, R. S.; Ingram, D. Studies of protoplast culture and plant regeneration from commercial and rapid-cycling Brassica species. Plant Cell Tiss. Organ Cult. 19:213–224; 1989.

    Article  Google Scholar 

  • Murashige, T.; Skoog, F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 15:473–497; 1962.

    Article  CAS  Google Scholar 

  • Narasimhulu, S. B.; Kirti, P. B.; Mohapatra, T.; Prakash, S.; Chopra, V. L. Shoot regeneration in stem explants and its amenability to Agrobacterium tumefaciens mediated gene transfer in Brassica carinata. Plant Cell Rep. 11:359–362; 1992.

    CAS  Google Scholar 

  • Paterson, K. E. Polarity of regeneration in excised leaves of Crassula argentea. 1. A role of auxin. Can. J. Bot. 61:1058–1063; 1983.

    CAS  Google Scholar 

  • Pua, E. C. Cellular and molecular aspects of ethylene on plant morphogenesis of recalcitrant Brassica species in vitro. Bot. Bull. Acad. Sinica 34:191–209; 1993.

    CAS  Google Scholar 

  • Bood, S.; Hedden, P. Convergent pathways of gibberellin A1 biosynthesis in Brassica. Plant Growth Reg. 15:241–246; 1994.

    Article  Google Scholar 

  • Stringam, G. R. Regeneration in stem explants of haploid rapessed (Brassica napus L.) Plant Sci. Lett. 9:115–119; 1977.

    Article  CAS  Google Scholar 

  • Teo, W.; Lakshmanan, P.; Kumar, P.; Goh, C. J.; Swarup, S. Direct shoot formation and plant regeneration from cotyledon explants of rapid-cycling Brassica rapa. In Vitro Cell. Dev. Biol. Plant 33:288–292; 1997.

    Google Scholar 

  • Williams, P. H.; Hill, C. B. Rapid-cycling populations of Brassica. Science 232:1385–1390; 1986.

    Article  PubMed  Google Scholar 

  • Wong, R. S. C.; Zee, S. Y.; Swanson, E. B. Isolated microspore culture of Chinese flowering cabbage (Brassica campestris spp. parachinensis) Plant Cell Rep. 15:396–400; 1996.

    Article  CAS  Google Scholar 

  • Yang, M. Z.; Jia, S. R.; Pua E. C. High frequency of plant regeneration from hypocotyl explants of Brassica carinata. Plant Cell Tiss. Organ Cult. 24:79–82; 1991.

    Article  Google Scholar 

  • Zar, J. H. Biostatistical analysis 2nd edn. Englewood Cliffs, NJ: Prentice-Hall; 1984:718.

    Google Scholar 

Download references

Author information

Authors and Affiliations


Corresponding author

Correspondence to Sanjay Swarup.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Cheng, PK., Lakshmanan, P. & Swarup, S. High-frequency direct shoot regeneration and continuous production of rapid-cycling Brassica oleracea in vitro . In Vitro Cell.Dev.Biol.-Plant 37, 592–598 (2001).

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI:

Key words

  • rapid-cycling brassica
  • plant development
  • morphogenesis
  • ethylene
  • thidiazuron