Advertisement

Plant Growth Regulation

, 65:381 | Cite as

In vitro induction and proliferation of protocorm-like bodies (PLBs) from leaf segments of Phalaenopsis bellina (Rchb.f.) Christenson

  • A. A. Khoddamzadeh
  • U. R. SinniahEmail author
  • M. A. Kadir
  • S. B. Kadzimin
  • M. Mahmood
  • S. Sreeramanan
Original paper

Abstract

An in vitro culture procedure was established to induce protocorm-like bodies (PLBs) from leaf segments of the Phalaenopsis bellina (Rchb.f.) Christenson directly from epidermal cells without intervening callus on ½ strength modified Murashige and Skoog (MS) (in Physiol Plant 15:473–497, 1962) medium supplemented with 1-Naphthaleneacetic acid (NAA; 0, 0.1, 1 mg/l) and Thidiazuron (TDZ; 0, 0.1, 1, 3 mg/l). The best response was established at 3 mg/l TDZ which induced 78% of leaf segments to form a mean number of 14 PLBs per explant after 16 weeks of culture. No PLBs were found when leaf segments were cultured on ½ strength modified MS media supplemented with 0.1 and 1 mg/l NAA. The best induction percentage for auxin: cytokinin combination was at the combination of NAA and TDZ at 1.0 and 3.0 mg/l which gave 72% induction with 9 PLBs per explant. Semi-solid ½ strength MS and liquid Vacin and Went (VW) (in Bot Gaz 110:605–613, 1949) medium were used in order to find the highest survival and number of PLBs proliferation after 3 months in culture. Half strength MS showed an average of 9 PLBs in comparison with VW with an average of 5.3 PLBs per explants. Histological observations revealed that the regenerated PLBs were generally formed from the epidermal layers of the posterior regions of the leaf segments. Scanning electron micrograph of PLBs showed the origin of newly formed PLB from the peripheral region of leaf segments.

Keywords

Protocorm Direct organogenesis Monopodial orchid Naphthaleneacetic acid Thidiazuron 

Abbreviations

MS

Murashige and Skoog media

VW

Vacin and Went media

NAA

1-Naphthaleneacetic acid

TDZ

1-phenyl-3-(1, 2, 3-thiadiazol-5-yl)-urea

PLB

Protocorm like body

References

  1. Arditti J, Ernst R (1993) Micropropagation of orchids. Wiley Publishers, New YorkGoogle Scholar
  2. Bhadra SK, Hossain MM (2004) Induction of embryogenesis and direct organogenesis in Micropera pallida Lindl., an epiphytic orchid of Bangladesh. J Orchid Soc India 18:5–9Google Scholar
  3. Chen JT, Chang WC (2001) Effects of auxins and cytokinins on direct somatic embryogenesis on leaf explants of Oncidium ‘Gower Ramsey’. Plant Growth Regul 34:229–232CrossRefGoogle Scholar
  4. Chen JT, Chang WC (2004) Induction of repetitive embryogenesis from seed-derived protocorms of Phalaenopsis amabilis var. formosa Shimadzu. In Vitro Cell Dev Biol 40:290–293CrossRefGoogle Scholar
  5. Chen JT, Chang WC (2006) Direct somatic embryogenesis and plant regeneration from leaf explants of Phalaenopsis amabilis. Biol Plant 50:169–173CrossRefGoogle Scholar
  6. Chen Y, Piluek C (1995) Effects of thidiazuron and N6- benzylaminopurine on shoot regeneration of Phalaenopsis. Plant Growth Regul 16:99–101CrossRefGoogle Scholar
  7. Chen JT, Chang C, Chang WC (1999) Direct somatic embryogenesis on leaf explants of Oncidium ‘Gower Ramsey’ and subsequent plant regeneration. Plant Cell Rep 19:143–149CrossRefGoogle Scholar
  8. Chen YC, Chang C, Chang WC (2000) A reliable protocol for plant regeneration from callus culture of Phalaenopsis. In Vitro Cell Dev Biol Plant 36:420–423CrossRefGoogle Scholar
  9. Chugh S, Guha S, Rao RU (2009) Micropropagation of orchids: a review on the potential of different explants. Sci Hortic 122:507–520CrossRefGoogle Scholar
  10. Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42CrossRefGoogle Scholar
  11. Gersterberger P, Leins P (1978) Rasterelektronenmikroskopische Untersuchungen an Blütenknospen vonPhysalis philadelphica (Solanaceae)-Anwendung einer neuen Präparationsmethode. Ber Deutsch Bot Ges 91:381–387Google Scholar
  12. Hew CS, Yong JWH (2004) The physiology of tropical orchids in relation to the industry. New Jersey World Scientific Pub Co. Inc., NJGoogle Scholar
  13. Hsiao YY, Tsai WC, Kuoh CS, Huang TH, Wang HC, Wu TS, Leu YL, Chen WH, Chen HH (2006) Comparison of transcripts in Phalaenopsis bellina and Phalaenopsis equestris (Orchidaceae) flowers to deduce monoterpene biosynthesis pathway. BMC Plant Biol 6:14PubMedCrossRefGoogle Scholar
  14. Huetteman CA, Precee JE (1993) Thidiazuron: a potent cytokinin for woody plant tissue culture. Plant Cell Tissue Organ Cult 33:105–119CrossRefGoogle Scholar
  15. Islam MO, Rahman ARMM, Matsui S, Prodhan AKMA (2003) Effects of complex organic extracts on callus growth and PLB regeneration through embryogenesis in the Doritaenopsis orchid. Jpn Agric Res Q 37(4):229–235Google Scholar
  16. Jensen WA (1962) Botanical histochemistry. Freeman, San FranciscoGoogle Scholar
  17. Lin CC (1986) In vitro culture of flower stalk internodes of Phalaenopsis and Doritaenopsis. Lindleyana 1:158–163Google Scholar
  18. Mahmood M, Chew YC (2008) Agrobacterium-mediated genetic transformation of Phalaenopsis bellina using GFP and GUS reporter genes. Pertanika J Sci & Technol 16(2):129–139Google Scholar
  19. Mayr H (1998) Orchid names and their meaning. Koeltz Scientific Books, KoenigsteinGoogle Scholar
  20. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue culture. Physiol Plant 15:473–497CrossRefGoogle Scholar
  21. Nagl W, Rücker W (1976) Effects of phytohormones on thermal denaturation profiles of Cymbidium DNA: indication of differential DNA replication. Nucleic Acids Res 3:2033–2039PubMedGoogle Scholar
  22. Nash N (2003) Phalaenopsis primer: a beginner’s guide to growing moth orchids. Orchids 72:906–913Google Scholar
  23. Park SY, Murthy HN, Paek KY (2002a) Rapid propagation of Phalaenopsis from floral stalk-derived leaves. In Vitro Cell Dev Biol Plant 38:168–172CrossRefGoogle Scholar
  24. Park SY, Yeung EC, Chakrabarty D, Paek KY (2002b) An efficient direct induction of protocorm-like bodies from leaf subepidermal cells of Doritaenopsis hybrid using thin-section culture. Plant Cell Rep 21:46–51CrossRefGoogle Scholar
  25. Riera-Lizarazu O, Mujeeb-Kazi A, William MDHM (1992) Maize (Zea mays L.) mediated polyhaploid production in some Triticeae using a detached tiller method. J GenetBreed 46:335–346Google Scholar
  26. Saieed NT, Douglas GC, Fry DJ (1994) Induction and stability of somaclonal variation in growth, leaf phenotype and gas exchange characteristics of Poplar regenerated from callus culture. Tree Physiol 14:1–16PubMedGoogle Scholar
  27. Shiau YJ, Nalawade SM, Hsia CN, Mulabacal V, Tsay H (2005) In vitro propagation of the Chinese medicinal plant, Dendrobium candidum Wall. Ex Lindl., from axenic nodal segments. In Vitro Cell Dev Biol Plant 41:666–670CrossRefGoogle Scholar
  28. Subramanium G, Taha RM (2003) Morphogenesis of Cymbidium atropurpureum in vitro. Malays J Sci 22:1–5Google Scholar
  29. Talukder SK, Nasiruddin KM, Yesmin S, Hassan L, Begum R (2003) Shoot proliferation of Dendrobium orchid with BAP and NAA. J Biol Sci 3(11):1058–1062CrossRefGoogle Scholar
  30. Tanaka M, Sakanishi Y (1980) Clonal propagation of Phalaenopsis through tissue culture. In: Kashemsanta MRS (ed) Proceedings of the 9th world orchid conference, Bangkok, pp 215–221Google Scholar
  31. Tanaka M, Hasegawa A, Goi M (1975) Studies on the clonal propagation of monopodial orchids by tissue culture. I. Formation of protocorm-like bodies from leaf tissues in Phalaenopsis and Vanda. J Jap Soc Hort Sci 44:47–58CrossRefGoogle Scholar
  32. Teob ES (1989) Orchids of Asia. Times Books International, Singapore, pp 125–134Google Scholar
  33. Thammasiri K (2002) Preservation of seeds of some Thai orchid species by vitrification. In: Proceedings of the 16th world orchid conference, pp 248–251Google Scholar
  34. Tokuhara K, Mii M (2003) Highly-efficient somatic embryogenesis from cell suspension cultures of Phalaenopsis orchids by adjusting carbohydrate sources. In Vitro Cell Dev Biol 39:635–639CrossRefGoogle Scholar
  35. Vacin E, Went F (1949) Some pH changes in nutrient solutions. Bot Gaz 110:605–613CrossRefGoogle Scholar
  36. Vajrabhaya M, Vajrabhaya T (1970) Culture of Rhynchostylis gigantea a monopodial orchid. Am Orchid Soc Bull 39:907–910Google Scholar
  37. Ye XL, Cheng SJ, Wang FX, Qian NF (1988) Morphology of immature seeds and development in vitro of Dendrobium candidum. Acta Bot Yunnanica 10:285–290Google Scholar
  38. Zhang QX, Fang YM (2005) Tissue culture and in vitro seedling and protocorm-like body examination of Dendrobium candidum. Acta Bot Boreali- Occidentalia Sinica 25:1761–1765Google Scholar
  39. Zhao P, Wu F, Feng FS, Wand WJ (2008) Protocorm-like body (PLB) formation and plant regeneration from the callus culture of Dendrobium candidum wall ex Lindl. In Vitro Cell Dev Bio Plant 44:178–185CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • A. A. Khoddamzadeh
    • 1
  • U. R. Sinniah
    • 1
    Email author
  • M. A. Kadir
    • 2
  • S. B. Kadzimin
    • 1
  • M. Mahmood
    • 3
  • S. Sreeramanan
    • 4
  1. 1.Department of Crop Science, Faculty of AgricultureUniversiti Putra Malaysia (UPM)SerdangMalaysia
  2. 2.Department of Agrotechnology, Faculty of AgricultureUniversiti Putra Malaysia (UPM)SerdangMalaysia
  3. 3.Department of Biochemistry, Faculty of Biotechnology and Biomolecular SciencesUniversiti Putra Malaysia (UPM)SerdangMalaysia
  4. 4.School of Biological SciencesUniversiti Sains Malaysia (USM)GeorgetownMalaysia

Personalised recommendations