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In vitro shoot induction and plant regeneration from flower buds in Paphiopedilum orchids

  • Plant Tissue Culture
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Abstract

Paphiopedilum species are recalcitrant in tissue culture, and no explant from mature plants has been successfully mass propagated in vitro. This study was aimed at inducing shoots and regenerating plants from the flowering plants of a sequentially flowering Paphiopedilum Deperle and a single floral Paphiopedilum Armeni White. By using cross-sectioned flower buds (FBs), we found that in both species, only sections that contained the base tissue of FBs were able to produce shoots and plants. We have also found that sections of FBs between 1.5 and 3.0 cm from Paphiopedilum Deperle were able to produce shoots, but only sections of FBs >2.5 cm from Paphiopedilum Armeni White were regenerable. Our microscopic observations revealed that the small bract at the FB base harbored a new miniature FB, which further harbored a primitive FB with dome-shaped meristem-like tissues that presumably led to the plant induction. The reiteration of this pattern resulted in a scorpioid cyme inflorescence architecture in the multifloral Paphiopedilum species, and its failure to reiterate resulted in a single flower. The induction rates were 57–75%, and all plants survived in a greenhouse. This method is potentially applicable for the micropropagation and conservation of slipper orchids.

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References

  • Airoldi C. A. Determination of sexual organ development. Sex. Plant Reprod. 23: 53–62; 2010.

    Article  PubMed  Google Scholar 

  • Arditti J.; Clements M. A.; Fast G.; Hadley G.; Nishimura G.; Emst R. Orchid seed germination and seedling culture—a manual. In: Arditti J. (ed) Orchid biology reviews and perspectives, II. Cornell University Press, Ithaca, pp 243–370; 1982.

    Google Scholar 

  • Arditti J.; Ernst R. Micropropagation of orchids. Wiley, New York, pp 467–520; 1993.

    Google Scholar 

  • Chen T. Y.; Chen J. T.; Chang W. C. Plant regeneration through direct shoot bud formation from leaf cultures of Paphiopedilum orchids. Plant Cell Tiss. Org. Cult. 76: 11–15; 2004.

    Article  CAS  Google Scholar 

  • Chen W. H.; Chen T. M.; Fu Y. M.; Hsieh R. M.; Chen W. S. Studies on somaclonal variation in Phalaenopsis. Plant Cell Rep. 18: 7–13; 1998.

    Article  Google Scholar 

  • Chugh S.; Guha S.; Rao I. U. Micropropagation of orchids: A review on the potential of different explants. Sci. Hortic. 122: 507–520; 2009.

    Article  CAS  Google Scholar 

  • CITES (2008) Convention on International Trade in Endangered Species of Wild Fauna and Flora Appendices I, II and III. Geneva, Switzerland. http://www.cites.org/eng/app/E-Jul01.pdf.

  • De Pauw M.; Remphrey W. R. In vitro germination of three Cypripedium species in relation to time of seed collection, media, and cold treatment. Can. J. Bot. 71: 879–885; 1993.

    Article  Google Scholar 

  • Duckhouse H.; Mason T. J.; Phull S. S.; Lorimer J. P. The effect of sonication on microbial disinfection using hypochlorite. Ultrason. Sonochem. 11: 173–176; 2004.

    Article  PubMed  CAS  Google Scholar 

  • Hong P. I.; Chen J. T.; Chang W. C. Plant regeneration via protocorm like body formation and shoot multiplication from seed derived callus of a maudiae type slipper orchid. Acta. Physiol. Plant 30: 755–759; 2008.

    Article  CAS  Google Scholar 

  • Huang L. C. A procedure for asexual multiplication of Paphiopedilum in vitro. Am. Orchid Soc. Bull. 57: 274–278; 1988.

    Google Scholar 

  • Huang L. C.; Lin C. J.; Kuo C. I.; Huang B. L.; Murashige T. Paphiopedilum cloning in vitro. Sci. Hortic. 91: 111–121; 2001.

    Article  Google Scholar 

  • Koopowitz H. Tropical slipper orchids: Paphiopedilum and Phragmipedium species and hybrids. Timber Press, Portland, p 41; 2008.

    Google Scholar 

  • Lauzer D.; St-Arnaud M.; Barabé D. Tetrazolium staining and in vitro germination of mature seeds of Cypripedium acaule (Orchidaceae). Lindleyana 9: 197–204; 1994.

    Google Scholar 

  • Lin Y. H.; Chang C.; Chang W. C. Plant regeneration from callus culture of a Paphiopedilum hybrid. Plant Cell Tiss. Org. Cult. 62: 21–25; 2000.

    Article  CAS  Google Scholar 

  • Long B.; Niemiera A. X.; Cheng Z. Y.; Long C. L. In vitro propagation of four threatened Paphiopedilum species (Orchidaceae). Plant Cell Tiss. Org. Cult. 101: 151–162; 2010.

    Article  Google Scholar 

  • McSteen P.; Leyser O. Shoot branching. Ann. Rev. of Plant Biol. 56: 353–374; 2005.

    Article  CAS  Google Scholar 

  • Murashige T.; Skoog F. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 495–497; 1962.

    Article  Google Scholar 

  • Ng C.-Y.; Saleh N. M. In vitro propagation of Paphiopedilum orchid through formation of protocorm-like bodies. Plant Cell Tiss. Org. Cult. 105: 193–202; 2010.

    Article  Google Scholar 

  • Ng C.-Y.; Saleh N. M.; Zaman F. Q. In vitro multiplication of the rare and endangered slipper orchid, Paphiopedilum rothschildianum (Orchidaceae). African Journal of Biotechnology 9: 2062–2068; 2010.

    CAS  Google Scholar 

  • Philip V. J.; Nainar S. A. Z. Clonal propagation of Vanilla planifolia (Salisb.) Ames using tissue culture. J. Plant Physiol. 122: 211–215; 1986.

    Google Scholar 

  • Prusinkiewicz P.; Erasmus Y.; Lane B.; Lawrence D.; Harder L. D.; Coen E. Evolution and development of inflorescence architectures. Science 316: 1452–1456; 2007.

    Article  PubMed  CAS  Google Scholar 

  • Santos Diaz Md. S.; Álvare C. C. Plant regeneration through direct shoot formation from leaf cultures and from protocorm-like bodies derived from callus of Encyclia mariae (Orchidaceae), a threatened Mexican orchid. In Vitro Cell Dev. Biol. Plant 45: 162–170; 2009.

    Article  Google Scholar 

  • Stewart J.; Button J. Tissue culture studies in Paphiopedilum. Am. Orchid Soc. Bull. 44: 591–599; 1975.

    Google Scholar 

  • Teob E. S. Orchids of Asia. Times Books International, Singapore, pp 125–134; 1989.

    Google Scholar 

  • Tomita M.; Tomita M. Plant regeneration from immature seed-derived callus of Cypripedium macranthos Swartz var. taiwanianum (Masamune) F. Maekawa. Breed. Sci. 47: 279–281; 1997.

    Google Scholar 

  • U. S. Department of Agriculture. Floriculture crops 2008 summary. National Agricultural Statistics Service, Washington, DC; 2009.

    Google Scholar 

  • Whigham D. F.; Neill J. P. O.; Rasmussen H. N.; Caldwell B. A.; McCormick M. K. Seed longevity in terrestrial orchids—potential for persistent in situ seed banks. Biol. Conserv. 129: 24–30; 2006.

    Article  Google Scholar 

  • Yam T. W.; Arditti J. History of orchid propagation: a mirror of the history of biotechnology. Plant Biotechnol. Rep. 3: 1–56; 2009.

    Article  Google Scholar 

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Acknowledgment

We thank Dr. Michael Fine of Virginia Commonwealth University for reviewing the manuscript and for advice on data analysis.

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Correspondence to Fang-Sheng Wu.

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Editor: D. Tomes

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Liao, YJ., Tsai, YC., Sun, YW. et al. In vitro shoot induction and plant regeneration from flower buds in Paphiopedilum orchids. In Vitro Cell.Dev.Biol.-Plant 47, 702–709 (2011). https://doi.org/10.1007/s11627-011-9370-7

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  • DOI: https://doi.org/10.1007/s11627-011-9370-7

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