In Vitro Cellular & Developmental Biology - Plant

, Volume 46, Issue 6, pp 537–548 | Cite as

An elite protocol for accelerated quality-cloning in Gerbera jamesonii Bolus cv. Sciella

  • Saikat Gantait
  • Nirmal Mandal
  • Somnath Bhattacharyya
  • Prakash Kanti Das
Micropropagation

Abstract

A novel, efficient, and simple protocol was developed on in vitro mass propagation and acclimatization of Gerbera jamesonii Bolus cv. Sciella, an ornamental plant with attractive flowers. Shoot tip was used as the primary explant for in vitro establishment in which Murashige and Skoog (MS) medium supplemented with a low level of NAA (0.5 mg l−1) and BAP (1.5 mg l−1) promoted earliest axillary bud initiation within 5 d in 91.6% of the inoculants. Five axillary buds were initiated from a single explant within 13 d after inoculation. A very high rate of shoot multiplication (14 shoots per inoculated axillary bud) and proliferation was achieved when MS medium was fortified with a relatively higher level of BAP (2 mg l−1) and 60 mg l−1 ADS within 27 d of multiple shoot culture. A maximum number of well-developed roots per plant was observed in MS medium with 0.5 mg l−1 IAA in the next 26 d. In the easy low-cost acclimatization process of 20 d, a combination of sand, soil, cow urine, and tea leaves extract (1:1:1:1; v/v) ensured 95% survival rate. Sixty-one well-acclimatized plants were obtained from a single shoot tip within 86 d. The sustained multiple shoot culture for 15 mo paved the way toward the conservation of genetic resources as well as beneficial economics. The clonal fidelity study of micropropagated and sustained cultured clones using ISSR primers ensured the continuous supply of quality propagules retaining genetic uniformity. The in vitro-generated plants performed better over conventionally propagated plants in the field condition.

Keywords

Acclimatization Commercial scale Field performance Gerbera ISSR Micropropagation Sustained culture 

References

  1. Alderete L. M.; Mori M.; Kato A.; Escandón A. S. Establishment of an in vitro micropropagation protocol for Mecardonia tenella. Electron J Biotechnol 9: 263–266; 2006.CrossRefGoogle Scholar
  2. Aswath C. R.; Choudhary M. L. Rapid plant regeneration from Gerbera jamesonii Bolus callus cultures. Acta Bot Croat 61: 125–134; 2002.Google Scholar
  3. Barbosa M. H. P.; Pasqual M.; Pinto J. E. B. P.; Pinto C. A. B. P. In vitro propagation of Gerbera jamesonii Bolus ex Hock cv. Applebloesem: Effect of adenine, tyrosine and salts concentrations of MS medium. Cienc Prat 17: 151–154; 1993.Google Scholar
  4. Bhatia R.; Sing K. P.; Jhang T.; Sharma T. R. Assessment of clonal fidelity of micropropagated gerbera plants by ISSR markers. Sci Hortic 119: 208–211; 2009.CrossRefGoogle Scholar
  5. Bose TK, Yadav LP (1993) Commercial flowers. Naya Prakash. 206 Bidhan Sarani, Kolkata. India: 601–622Google Scholar
  6. Chattopadhyay K.; Bhattacharyya S.; Mandal N.; Sarkar H. K. PCR-based characterization of mungbean (Vigna radiata) genotypes from Indian subcontinent at intra- and inter-specific level. J Plant Biochem Biotech 17: 141–148; 2008.Google Scholar
  7. Duncan D. B. Multiple range and multiple F test. Biometrics 11: 1–42; 1955.CrossRefGoogle Scholar
  8. Gantait S. Accelerated cloning in vitro and induction of variation in six plants having medicinal, aromatic or aesthetic importance. PhD Thesis, Bidhan Chandra Krishi Viswavidyalaya, West Bengal., India: 166; 2009Google Scholar
  9. Gantait S.; Mandal N. Tissue culture of Anthurium anderanum: a significant review and future prospective. Int J Bot 6: 207–219; 2010.Google Scholar
  10. Gantait S.; Mandal N.; Bhattacharyya S.; Das P. K. In vitro mass multiplication with pure genetic identity in Anthurium andreanum Lind. Plant Tissue Cult Biotech 18: 113–122; 2008.Google Scholar
  11. Gantait S.; Mandal N.; Bhattacharyya S.; Das P. K. In vitro mass multiplication with genetic clonality in elephant garlic (Allium ampeloprasum L.). J Crop Weed 5: 100–104; 2009.Google Scholar
  12. George EF (1996) Plant propagation by tissue culture. Part 2. Exegetics Ltd., Edington, Westbury, Wilts, England UKGoogle Scholar
  13. George P. S.; Ravishankar G. A. In vitro multiplication of Vanilla planifolia using axillary bud explants. Plant Cell Rep 16: 490–494; 1997.Google Scholar
  14. Gustavsson B. A.; Stanys V. Field performance of sanna lingonberry derived by micropropagation vs stem cutting. Hortic Sci 35: 742–744; 2000.Google Scholar
  15. Henrique M.; Barbosa P.; Pinto J. E. B. P.; Pinto C. A. B. P.; Innecco R. In vitro propagation of Gerbera jamesonii Bolus ex Hook Appelbloesum using young capitilum. Rev Ceres 41: 386–395; 1994.Google Scholar
  16. Huang H.; Li L.; Shenghui S. Tissue culture of Gerbera jamesonii Bolus. Jishou Daxue Xuebao Ziran Kexueban 22: 35–41; 2001.Google Scholar
  17. Huang J. H. In vitro propagation and preservation of ginger germplasm resources. Sci Agri Sin 28: 24–30; 1995.Google Scholar
  18. Jerzy M.; Lubomski M. Adventitious shoot formation on ex vitro derived leaf explants of Gerbera jamesonii. Sci Hortic 47: 115–124; 1991.CrossRefGoogle Scholar
  19. Murashige T. Plant tissue culture and its biotechnological application. In Baez WE, Reinhard E, Zenk MH (eds) Springer, Amerdam, The Netherlands, 392–403; 1976Google Scholar
  20. Murashige T.; Skoog F. A revised medium for rapid growth and bio-assays with tobacco tissue cultures. Physiol Plant 15: 473–497; 1962.CrossRefGoogle Scholar
  21. Naik D.; Vartak V.; Bhargava S. Provenance and subculture-dependent variation during micropropagation of Gmelina arborea. Plant Cell Tissue Organ Cult 73: 189–195; 2003.CrossRefGoogle Scholar
  22. Orlikowska T.; Nowak E.; Marasek A.; Kucharska D. Effects of growth regulators and incubation period on in vitro regeneration of adventitious shoots from gerbera petioles. Plant Cell 59: 95–102; 1999.Google Scholar
  23. Parthasarathy V. A.; Nagaraju V. In-vitro propagation in Gerbera jemesonii Bolus. Ind J Hort 56: 82–85; 1999.Google Scholar
  24. Ray T.; Dutta I.; Saha P.; Das S.; Roy S. C. Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of Indo-Ganjetic plains, as assessed by RAPD and ISSR markers. Plant Cell Tissue Organ Cult 85: 11–21; 2006.CrossRefGoogle Scholar
  25. Ray T.; Saha P.; Roy S. C. In vitro plant regeneration from young capitulum explants of Gerbera jamesonii. Plant Cell Biotech Mol Biol 6: 35–40; 2005.Google Scholar
  26. Reynoird J. P.; Chriqui D.; Noin M.; Brown S.; Marie D. Plant regeneration from in vitro leaf culture of several gerbera species. Plant Cell Tissue Organ Cult 33: 203–210; 1993.CrossRefGoogle Scholar
  27. Ruffoni B.; Massabo F. Tissue culture in Gerbera jamesonii hybrida. Acta Hortic 289: 147–148; 1991.Google Scholar
  28. Sakila S.; Ahmed M. B.; Roy U. K.; Biswas M. K.; Karim R.; Razvy M. A.; Hossain M.; Islam R.; Hoque A. Micropropagation of strawberry (Fragaria x ananassa Duch.) a newly introduced crop in Bangladesh. Am-Eura J Sci Res 2: 151–154; 2007.Google Scholar
  29. Schiva T. Annali dell. Istituto Sperimentale per la Floricaltura 6: 133–135; 1975.Google Scholar
  30. Sharry E. S.; Teixeira da Silva J. A. Effective organogenesis, somatic embryogenesis and salt tolerance induction in vitro in the Persian Lilac tree (Melia azedarach L.). In: da Silva JA Teixeira (ed) Floriculture, ornamental and plant biotechnology: Advances and topical issues (vol 2). Global Science, Isleworth, pp 317–324; 2006.Google Scholar
  31. Thomas P.; Ravindra M. B. Effect of pruning or removal of in vitro formed roots on ex vitro root regeneration and growth in micropropagated grapes. Plant Cell Tissue Organ Cult 51: 177–180; 1997.CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2010

Authors and Affiliations

  • Saikat Gantait
    • 1
  • Nirmal Mandal
    • 1
  • Somnath Bhattacharyya
    • 2
  • Prakash Kanti Das
    • 2
  1. 1.Department of Biotechnology, Instrumentation and Environmental ScienceBidhan Chandra Krishi ViswavidyalayaMohanpurIndia
  2. 2.Department of GeneticsBidhan Chandra Krishi ViswavidyalayaMohanpurIndia

Personalised recommendations