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Plant Cell, Tissue and Organ Culture

, Volume 91, Issue 3, pp 243–250 | Cite as

TDZ-induced high frequency plant regeneration through multiple shoot formation in witloof chicory (Cichorium intybus L.)

  • Buhara Yucesan
  • Arzu Ucar Turker
  • Ekrem GurelEmail author
Original Paper

Abstract

A very efficient and rapid regeneration system via multiple shoot formation was developed for Cichorium intybus L. when leaf explants excised from sterile seedlings were cultured on medium supplemented with different concentrations and combinations of various plant growth regulators. In a comparison of leaf lamina and petiole explants, lamina explants produced over three times more shoots than petiole explants, with a mean of 7.5 shoots compared to 2.4. Of the combinations of KIN/IAA, KIN/NAA, BAP/IAA, or BAP/NAA, 0.5 mg l−1 KIN combined with 0.3 mg l−1 IAA was the most effective, producing a mean of 19.7 shoots per lamina explant while the control treatment involving no plant growth regulators produced no shoots at all. When either cytokinin was used alone, BAP was found nearly twice more successful than KIN. However, the most effective treatment of all was the combination of 0.01 mg l−1 TDZ and 1.0 mg l−1 IAA, producing as many as 35.8 shoots per lamina explant. This rate of shoot regeneration is remarkably higher than those previously reported for C. intybus, most likely due to the highly inductive effect of TDZ, which was tested for the first time in this species. Rooting of the shoots was readily achieved on medium containing different concentrations of IAA or IBA. IAA was more effective than IBA and resulted in the highest frequency of shoots that rooted (100%) and mean number of roots per shoot (4.2) when used at 0.5 mg l−1. Hardening off process resulted in a production of more than 80% healthy plantlets.

Keywords

Cichorium intybus L. Lamina explants Petiole explants Multiple shoot formation In vitro regeneration TDZ 

Notes

Acknowledgements

The authors are grateful to The Scientific and Technological Research Council of Turkey (TUBITAK) for financial support [TBAG-2278(103T024) and TBAG-HD/27(105T040)].

References

  1. Bais HP, Ravishankar GA (2001) Cichorium intybus L—cultivation, processing, utility, value addition and biotechnology, with an emphasis on current status and future prospects. J Sci Food Agric 18:467–484CrossRefGoogle Scholar
  2. Belletre A, Coullerot JP, Vasseur J (1999) Effects of glycerol on somatic embryogenesis in Cichorium leaves. Plant Cell Rep 19:26–31CrossRefGoogle Scholar
  3. Bennici A, Schiff S, Mori B (2006) Morphogenic effect of colchicine in Cichorium intybus L. root explants cultured in vitro. Caryologia 59:284–290Google Scholar
  4. Charriére F, Sotta B, Emile M, Günther H (1999) Induction of adventitious shoots or somatic embryos on in vitro cultured zygotic embryos of Helianthus annuus: variation of endogenous hormone levels. Plant Physiol Biochem 37:751–757CrossRefGoogle Scholar
  5. Dandekar AM, Fisk H (2005) Plant transformation: Agrobacterium-mediated gene transformation. In: Pena L (ed) Transgenic plants: methods and protocols. Humana Press, New Jersey, pp 35–46Google Scholar
  6. Davis PH (1975) Flora of Turkey and the East Aegean Islands, vol 5. Edinburgh University Press, Edinburhg, UK, p 627Google Scholar
  7. Demeulemeester MAC, Voet A, De Profit MP (1995) Induction of stem elongation on in vitro chichory root explants under unfavorable photoperiodic conditions by giberellin A3. Plant Growth Regul 16:239–241CrossRefGoogle Scholar
  8. Gurel E, Wren MJ (1995) In vitro development from leaf explants of sugar beet (Beta vulgaris L.)—rhizogenesis and the effect of sequential exposure to auxin and cytokinin. Ann Bot 75:31–38CrossRefGoogle Scholar
  9. Heutteman CA, Preece JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tiss Org Cult 33:105–119CrossRefGoogle Scholar
  10. Jones MPA, Yi ZJ, Murch SJ, Saxena PK (2007) Thidiazuron-induced regeneration of Echinacea purpurea L.: micropropagation in solid and liquid culture systems. Plant Cell Rep 26:13–19PubMedCrossRefGoogle Scholar
  11. Karam NS, Al-Majathoub M (2000) Direct shoot regeneration and microtuberization in wild Cyclamen persicum Mill. using seedling tissue. Sci Hort 86:235–246CrossRefGoogle Scholar
  12. Linden L, Riikonen A (2006) Effects of 6-benzylaminopurine, thidiazuron and type of explant on in vitro shoot development of Acer platanoides L. Propag Ornam Plants 6:201–204Google Scholar
  13. MixWagner G, Eneva T (1996) Positive effect of cefotaxime on plant regeneration from Cichorium intybus L. leaf material. Landbauforsch Volk 46:166–168Google Scholar
  14. Mohamed-Yasseen M, Splittstoesser WE (1995) Somatic embryogenesis in witloof chicory through leaf suspension culture. Plant Cell Rep 14:804–806Google Scholar
  15. Murashige T, Skoog FA (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  16. Park EJ, Lim HT (1999) Establishment of an efficient in vitro plant regeneration system in chicory (Cichorium intybus L. var sativus). Acta Hort 483:367–370Google Scholar
  17. Patel VK, Bhatt HV (1985) Toxicity and antiseptic effect of chicory root extract in pyorrea. Antiseptic 31:904–906Google Scholar
  18. Petrovic J, Stanojkovic A, Comic L, Curcic S (2004) Antibacterial activity of Cichorium intybus. Fitoterapia 75:737–739PubMedCrossRefGoogle Scholar
  19. Pieron S, Belaizi M, Boxus P (1993). Nodule culture, a possible morphogenetic pathway in Cichorium intybus L. propagation. Sci Hort 53:1–11CrossRefGoogle Scholar
  20. Profumo P, Gastaldo P, Caffaro L, Dameri RM, Michelozzi GR, Bennici A (1985) Callus induction and plantlet regeneration in Cichorium intybus L.: II. effect of different hormonal treatments. Protoplasma 126:215–220CrossRefGoogle Scholar
  21. Rehman RU, Israr M, Srivastava PS, Bansal KC, Abdin MZ (2003) In vitro regeneration of witloof chichory (Cichorium intybus L.) from leaf explants and accumulation of esculin. In vitro Cell Dev Biol Plant 39:142–146CrossRefGoogle Scholar
  22. Trigiano RN, Gray DJ (2000) Plant tissue culture: concepts and laboratory exercises, 2nd edn. CRC PressGoogle Scholar
  23. Vesseur J, Dubois J, Hilbert JL, Couillerot JP (1995) Somatic embryogenesis in chichory (Cichorium intybus L.). In: Bajaj YPS (ed) Biotechnology and agricultural forestry, vol 31, somatic embryogenesis and synthetic seeds II. Springer, Berlin, pp 125–137Google Scholar
  24. Vermeulen A, Vaucheret H, Pautot V, Chupeau Y (1992) Agrobacterium-mediated transfer of a mutant Arabidopsis acetolactate synthase gene confers resistance to chlorsulfuron in chicory (Cichorium intybus L.). Plant Cell Rep 11:243–247CrossRefGoogle Scholar
  25. Wang J, Bao MZ (2007) Plant regeneration of pansy (Viola wittrockiana) ‘Caidie’ via petiole-derived callus. Sci Hort 111:266–270CrossRefGoogle Scholar
  26. Zafar R, Ali SM (1998) Anti-hepatotoxic effects of root and callus extracts of Cichorium intybus L. J Ethnopharmacol 63:227–231PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Buhara Yucesan
    • 1
  • Arzu Ucar Turker
    • 1
  • Ekrem Gurel
    • 1
    • 2
    Email author
  1. 1.Department of Biology, Faculty of Science and LiteratureAbant Izzet Baysal UniversityBoluTurkey
  2. 2.Department of Plant and Microbial BiologyUniversity of California BerkeleyBerkeleyUSA

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