Skip to main content
SpringerLink
Log in

Rapid shoot regeneration from thin cell layer explants excised from petioles and hypocotyls in four cultivars of Brassica napus L.

  • Original Paper
  • Published:
Plant Cell, Tissue and Organ Culture Aims and scope Submit manuscript

Abstract

The present work describes a procedure that allows for the easy and rapid induction of caulogenesis in four cultivars of Brassica napus L. from transversal Thin Cell Layers (tTCLs). In order to investigate the regeneration ability of this crop, the effects of genotype, explant source and culture medium were examined on shoot regeneration. The tTCL explants were excised from hypocotyl and petiole of 2-week-old seedlings and cultured on a solid basal MS medium supplemented with α-naphthaleneacetic acid (NAA: 0.1–0.4 mg l−1), 6-benzylamino-purine (BAP: 1–4 mg l−1) and sucrose (20–40 g l−1). A significant genotypic effect was observed between the four cvs; Jumbo and Drakkar displayed higher capacities to produce shoots than Pactol and Cossair. Regeneration commenced earlier and the percentage of shoot-producing explants as well as the number of shoots per regenerating explant was greater. The comparison between the regeneration ability of different explants showed that the hypocotyls exhibited a high rate of shoot organogenesis when they were cultured on MS medium supplemented with 3 mg l−1 BAP, 0.3 mg l−1 NAA and 30 g l−1 sucrose. Adventitious shoot buds developed from 46% of the tTCLs, with a mean of 7.5 buds. Furthermore, the method was fast with shoot formation occurring by 7 days culture. Plantlets regenerated from all shoots and developed normally. The regenerated plants were fertile and identical to source plants.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

BAP:

6-Benzylamino-purine

NAA:

α-Naphthaleneacetic acid

MS:

Murashige and Skoog’s medium (1962)

PGRs:

Plant growth regulators

tTCL(s):

Transverse thin cell layer(s)

References

  • Akasaka-Kennedy Y, Yoshida H, Takahata Y (2005) Efficient plant regeneration from leaves of rapeseed (Brassica napus L.): the influence of AgNO3 and genotype. Plant Cell Rep 24:649–654

    Article  PubMed  CAS  Google Scholar 

  • Bui VL, Nhut DT, Tran Thanh Van K (1999) Plant production via shoot regeneration from thin cell layer pseudo-bulblets explants of Lilium longiflorum in vitro. C R Acad Sci 322:303–310

    Google Scholar 

  • Charest PJ, Holbrook LA, Gabard J, Iyer VN, Miki BL (1988) Agrobacterium-mediated transformation of thin cell layer explants from Brassica napus L. Theor Appl Genet 75:438–445

    Article  Google Scholar 

  • Julliard J, Sossountzov L, Habricot Y, Pelletier G (1992) Hormonal requirement and tissue competency for shoot organogenesis in two cultivars of Brassica napus. Physiol Plant 84:521–530

    Article  CAS  Google Scholar 

  • Khehra GS, Mathias RJ (1992) The interaction of genotype, explant and media on the regeneration of shoots from complex explants of Brassica napus. J Exp Bot 43:1413–1418

    Article  Google Scholar 

  • Klimaszewska K, Keller WA (1985) High frequency plant regeneration from thin cell layer explants of Brassica napus. Plant Cell Tissue Organ Cult 4:183–197

    Article  CAS  Google Scholar 

  • Léguillon S, Charles G, Branchard M (2003) Plant regeneration from thin cell layers in Spinacia oleracea. Plant Cell Tissue Organ Cult 74:257–265

    Article  Google Scholar 

  • Mulin M, Bellio-Spataru A (2000) Organogenesis from hypocotyl thin cell layers of Lupinus mutabilis and Lupinus albus. Plant Growth Regul 30:177–183

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Narasimhulu SB, Chopra VL (1988) Species specific shoot regeneration response of cotyledonary explants of Brassicas. Plant Cell Rep 7:104–106

    Article  Google Scholar 

  • Nhut DT, Bui VL, Tran Thanh Van K (2000) Somatic embryogenesis and direct shoot regeneration of rice (Oryza sativa L.) using thin cell layer culture of apical meristematic tissue. J Plant Physiol 157:559–565

    CAS  Google Scholar 

  • Nhut DT, Bui VL, Fukai S, Tanaka M, Tran Thanh Van K (2001) Effects of activated charcoal, explant size, explant position and sucrose concentration on plant and shoot regeneration of Lilium longiflorum via young stem culture. Plant Growth Regul 33:59–65

    Article  Google Scholar 

  • Nhut DT, Huong NDT, Bui VL, Teixeira da Silva JA, Fukai S, Tanaka M (2002) The change in shoot regeneration potential of protocorm-like bodies derived from Lilium longiflorum young stem explant exposed to medium volume, pH, light intensity and sucrose concentration pretreatment. J Hortic Sci Biotechnol 77:79–82

    Google Scholar 

  • Nhut DT, Teixeira da Silva JA, Le BV, Tran Thanh Van K (2003) Thin cell layer studies of vegetable, leguminous and medicinal plants, Chapter 10. In: Nhut DT, Tran Thanh Van K, Le BV, Thorpe T (eds) Thin cell layer culture system: regeneration and transformation applications. Kluwer Academic Publishers, Dordrecht, pp 387–426

    Google Scholar 

  • O’Neill CM, Arthur AE, Mathias RJ (1996) The effects of proline, thioproline and methyglyoxal-bis-(guanylhydrazone) on shoot regeneration frequencies from stem explants of B. napus. Plant Cell Rep 15:695–698

    Article  CAS  Google Scholar 

  • Takasaki T, Hatakeyama K, Ojima K, Watanabe M, Toriyama K, Hinata K (1997) Factors influencing Agrobacterium-mediated transformation of Brassica rapa L. Breed Sci 47:127–134

    Google Scholar 

  • Tang GX, Zhou WJ, Li HZ, Mao BZ, He ZH, Yoneyama K (2003) Medium, explant and genotype factors influencing shoot regeneration in Oilseed Brassica spp. J Agron Crop Sci 189:351–358

    Article  Google Scholar 

  • Teixeira da Silva JA (2003) Thin cell layer technology in ornamental plant micropropagation and biotechnology. Afr J Biotechnol 2:683–691

    CAS  Google Scholar 

  • Tran Thanh Van M (1973) In vitro control of de novo flower, bud, root and callus differentiation from excised epidermal tissues. Nature 246:44–45

    Article  Google Scholar 

  • Turgut K, Barghchi M, Scott R (1998) Efficient shoot regeneration and somatic embryogenesis from immature cotyledons of Brassica napus L. Plant Breed 117:503–504

    Article  Google Scholar 

Download references

Acknowledgements

The authors thank the National Institute of Agronomic Research (INRA) in Rennes (France) for furnishing the seeds of Jumbo and the Tunisian National Institute of Agronomic Research (INRAT) in Tunis (Tunisia) for those of Drakkar, Pactol and Cossair.

Author information

Authors and Affiliations

  1. Laboratoire de Biotechnologie et Physiologie Végétales, ISAMOR/ESMISAB-Université de Bretagne Occidentale, Technopôle Brest-Iroise, Plouzane, 29280, France

    Asma Ben Ghnaya, Gilbert Charles & Michel Branchard

Authors
  1. Asma Ben Ghnaya
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gilbert Charles
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Michel Branchard
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gilbert Charles.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ben Ghnaya, A., Charles, G. & Branchard, M. Rapid shoot regeneration from thin cell layer explants excised from petioles and hypocotyls in four cultivars of Brassica napus L.. Plant Cell Tiss Organ Cult 92, 25–30 (2008). https://doi.org/10.1007/s11240-007-9298-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-007-9298-0

Keywords

Search

Navigation