Factors affecting micropropagation and acclimatization of an elite clone of Eucalyptus tereticornis Sm.

  • Diwakar Aggarwal
  • Anil Kumar
  • Jagriti Sharma
  • M. Sudhakara Reddy
Plant Tissue Culture

Abstract

Several factors influencing micropropagation of a selected elite clone of Eucalyptus tereticornis Sm. were investigated. Amongst different cytokinins tested, 6-benzyleadenine proved to be the most effective cytokinin for shoot multiplication and elongation. The initial size of the shoot clump (inoculum) also influenced shoot multiplication and elongation. The number of shoots proliferated per culture vessel were significantly higher (342 shoots per culture vessel) when larger shoot clumps (15–20 shoots) were inoculated, compared to smaller shoot clumps (4–5 shoots), which resulted in a reduced shoot proliferation rates (245 shoots per culture vessel). However, the number of elongated shoots (65 per culture vessel) and shoot length (5.23 cm) were higher in cultures which were inoculated with smaller shoot clumps in comparison to those cultures which were inoculated with larger shoot clumps (54 shoots per culture vessel with shoot length of 4.17 cm). The maximum number of rooted shoots (80.7 %) was obtained on one fourth-strength MS medium supplemented with 5.0 μM indolebutyric acid. The number of shoots proliferated, elongated, rooting frequency, and subsequent survival of plants after acclimatization were higher in cultures incubated under photosynthetically active radiation (PAR) compared to those incubated under cool fluorescent lights (CFL). Osmotic potential of the sap and chlorophyll content of cultures incubated under PAR were also higher than those incubated under CFL. Following transfer of plants to soil, inoculation with a suspension of Bacillus subtilis (plant growth-promoting bacterium) increased the survival rate of plants by 10 %, yielding successful transfer of 84 % of plants. Random amplified polymorphic DNA and inter simple sequence repeat analyses indicated a high level of clonal uniformity amongst regenerated plants and also with that of the mother plant.

Keywords

Biological hardening ISSR PAR light RAPD Shoot proliferation Tree micropropagation 

References

  1. Aggarwal D, Kumar A, Reddy MS (2010) Shoot organogenesis from elite plants of Eucalyptus tereticornis. Plant Cell Tissue Organ Cult 102:45–52CrossRefGoogle Scholar
  2. Appenroth KJ, Meço R, Jourdan V, Lillo C (2000) Phytochrome and post-translational regulation of nitrate reductase in higher plants. Plant Sci 159:51–56PubMedCrossRefGoogle Scholar
  3. Arnon DI (1949) Copper enzymes in isolated chloroplasts—polyphenol oxidases in Beta cularis. Plant Physiol 24:1–15PubMedCrossRefGoogle Scholar
  4. Beck SL, Dunlop RW (2001) Micropropagation of the Acacia species—a review. In Vitro Cell Dev Biol Plant 37:531–538CrossRefGoogle Scholar
  5. Bennett IJ, McComb JA, Tonkin CM, McDavid DAJ (1994) Alternating cytokinins in multiplication media stimulates In Vitro shoot growth and rooting of Eucalyptus globulus Labill. Ann Bot 74:53–58PubMedGoogle Scholar
  6. Bennett IJ, McDavid D, McComb JA (2003) The influence of ammonium nitrate, pH and indole butyric acid on root induction and survival in soil of micropropagated Eucalyptus globules. Biol Plant 47:355–360CrossRefGoogle Scholar
  7. Brainerd KE, Fuchigami LH (1982) Stomatal functioning of In Vitro and greenhouse apple leaves in darkness, mannitol, ABA and CO2. J Exp Bot 33:388–392CrossRefGoogle Scholar
  8. Brooker MIH (2000) A new classification of the genus Eucalyptus L’Her. (Myrtaceae). Aust Syst Bot 13:79–148CrossRefGoogle Scholar
  9. Chang SH, Donald DGM, Jacobs G (1992) Micropropagation of Eucalyptus radiata ssp. radiata using explants from mature and coppice material. S Afr For J 162:43–47Google Scholar
  10. Contin A, Van der Heijden R, Ten Hoopen HJG, Verpoorte R (1998) The inoculum size triggers tryptamine or secologanin biosynthesis in a Catharanthus roseus cell culture. Plant Sci 139:205–211CrossRefGoogle Scholar
  11. Dibax R, Eisfeld CL, Cuquel FL, Koehler H, Quoirin M (2005) Plant regeneration from cotyledonary explants of Eucalyptus camaldulensis. Sci Agric (Piracicaba, Brazil) 62:406–412CrossRefGoogle Scholar
  12. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
  13. Duncan DB (1955) Multiple range and multiple F tests. Biometrics 11:1–42CrossRefGoogle Scholar
  14. Figueiredo SFL, Simões C, Albarello N, Viana VRC (2000) Rollinia mucosa cell suspension cultures: establishment and growth conditions. Plant Cell Tissue Organ Cult 63:85–92CrossRefGoogle Scholar
  15. George EF (1996) Plant growth regulators. In: Plant propagation by tissue culture. Part 1. The Technology, 2nd ed. Exegetics, London, UK, pp 420–476Google Scholar
  16. Glocke P, Delaporte K, Collins G, Sedgley M (2006) Micropropagation of juvenile tissue of Eucalyptus erythronema × Eucalyptus stricklandii Cv. ‘Urrbrae Gem’. In Vitro Cell Dev Biol Plant 42:139–143CrossRefGoogle Scholar
  17. Goins GD, Yorio NC, Sanwoo MM, Brown CS (1997) Photomorphogenesis, photosynthesis and seed yield of wheat plants grown under red light-emitting diodes (LEDs) with and without supplemental blue lighting. J Exp Bot 48:407–413CrossRefGoogle Scholar
  18. Gomes F, Canhoto MJ (2003) Micropropagation of Eucalyptus nitens Maiden (Shining Gum). In Vitro Cell Dev Biol Plant 39:316–321CrossRefGoogle Scholar
  19. Hernandey-Sebastia C, Piehe Y, Desjardins Y (1999) Water relations of whole strawberry plantlets In Vitro inoculated with Glomus intraradices in a tripartite culture system. Plant Sci 143:81–91CrossRefGoogle Scholar
  20. Jain SM (2006) An update on overall recent progress on somatic embryogenesis in forest trees. In: Suzuki K, Ishii K, Sakurai S, Sasaki S (eds) Plantation technology in tropical forest science. Springer, Tokyo, pp 113–122CrossRefGoogle Scholar
  21. Jeon MW, Ali MB, Hahn EJ, Paek KY (2005) Effects of photon flux density on the morphology, photosynthesis and growth of a CAM orchid, Doritaenopsis during post-micropropagation acclimatization. Plant Growth Regul 45:139–147CrossRefGoogle Scholar
  22. Jones NB, Van Staden J (1997) Micropropagation of Eucalyptus. In: Bajaj YPS (ed) High-tech and micropropagation V, Biotechnology in agriculture and forestry, vol 39. Springer, Berlin Heidelberg New York, pp 286–329Google Scholar
  23. Khuspe SS, Gupta PK, Kulkarni DK, Mehta V, Mascarenhas AF (1987) Increased biomass production by Eucalyptus. Can J For Res 17:1361–1363CrossRefGoogle Scholar
  24. Kozai T (1991) Acclimatization of micropropagated plants. In: Bajaj YPS (eds) High-tech and micropropagation V, Biotechnology in agriculture and forestry, vol 39. Springer, Berlin Heidelberg New York, pp 127–141Google Scholar
  25. Kumar A, Aggarwal D, Gupta P, Reddy MS (2010) Factors affecting In Vitro propagation and field establishment of Chlorophytum borivillianum. Biol Plant 54:601–606Google Scholar
  26. Kumar A, Palni LMS, Nandi SK (2003) The effect of light source and gelling agent on micropropagation of Rosa damascene Mill. and Rhychostylis retusa (L.) Bl. J Hortic Sci Biotechnol 78:786–792Google Scholar
  27. Lal P (1993) Economics of clonal forestry plantation. In: Vivekanandhan K, Subramanian KN, Zabala NQ, Gurumurthy K (eds) Proceedings of workshop on production of genetically improved planting materials for afforestation programmes. RAS/91/004 field document no 7, Coimbatore, India, pp 108–115Google Scholar
  28. Lal P, Kulkarni HD, Srinivasa K (1993) Eucalyptus improvement program of ITC Bhadrachalam Paper Board Ltd. In: Vivekanandhan K, Subramanian KN, Zabala NQ, Gurumurthy K (eds) Proceedings of workshop on production of genetically improved planting materials for afforestation programmes. RAS/91/004 field document no 7, Coimbatore, India, pp 57–63Google Scholar
  29. Lee SH, Tewari RK, Hahn EJ, Paek KY (2007) Photon flux density and light quality induce changes in growth, stomatal development, photosynthesis and transpiration of Withania somnifera (L.) Dunal. Plantlets. Plant Cell Tissue Organ Cult 90:141–151CrossRefGoogle Scholar
  30. Lin C (2002) Blue light receptors and signal transduction. Plant Cell Suppl 14:S207–S225Google Scholar
  31. MacRae S, Van Staden J (1993) Agrobacterium rhizogenes mediated transformation to improve rooting ability of eucalypts. Tree Physiol 12:411–418PubMedGoogle Scholar
  32. Mullins KV, Llewellyn DJ, Hartney VJ, Strauss SH, Dennis ES (1997) Regeneration and transformation of Eucalyptus camaldulensis. Plant Cell Rep 16:787–791CrossRefGoogle Scholar
  33. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  34. Nugent G, Chandler SF, Whiteman P, Stevenson TW (2001a) Somatic embryogenesis in Eucalyptus globulus. Plant Cell Tissue Organ Cult 6:785–788Google Scholar
  35. Nugent G, Chandler SF, Whiteman P, Stevenson TW (2001b) Adventitious bud induction in Eucalyptus globulus Labill. In Vitro Cell Dev Biol Plant 37:388–391CrossRefGoogle Scholar
  36. Palni LMS, Bag N, Nadeem M, Tamta S, Vyas P, Bisht MS, Purohit VK, Kumar A, Nandi SK, Pandey A, Purohit AN (1998) Micropropagation: Conservation through tissue culture of selected Himalayan Plants. In: Agarwal DK, Farooquee NA, Nandi SK (eds) Research for mountain development: some initiatives and accomplishments. Gyanodaya Prakashan, Nainital, pp 431–452Google Scholar
  37. Pandey A, Bag N, Chandra B, Palni LMS (2002) Biological Hardening: A promising technology for tissue culture industry. In: Nandi SK, Palni LMS, Kumar A (eds) Role of plant tissue culture in biodiversity conservation and economic development. Gyanodaya Prakashan, Nainital, pp 565–577Google Scholar
  38. Pandey A, Palni LMS, Bag N (2000) Biological hardening of tissue culture raised tea plants. Biotechnol Lett 22:1087–1091CrossRefGoogle Scholar
  39. Pérez-Tornero O, Egea J, Vanoostende A, Burgos L (2000) Assessment of factors affecting adventitious shoot regeneration from In Vitro cultured leaves of apricot. Plant Sci 158:61–70PubMedCrossRefGoogle Scholar
  40. Pinto G, Park YS, Silva NL, Araújo C, Santos C (2008) Factors affecting maintenance, proliferation and germination of secondary somatic embryos of Eucalyptus globulus Labill. Plant Cell Tissue Organ Cult 95:69–78CrossRefGoogle Scholar
  41. Prakash MG, Gurumurthi K (2005) Somatic embryogenesis and plant regeneration in Eucalyptus tereticornis Sm. Curr Sci 88:1311–1316Google Scholar
  42. Prakash S, Van Staden J (2008) Micropropagation of Searsia dentate. In Vitro Cell Dev Biol Plant 44:338–341CrossRefGoogle Scholar
  43. Rani V, Raina SN (1998) Genetic analysis of enhanced-axillary branching-derived Eucalyptus tereticornis Smith and E camaldulensis Dehn. plants. Plant Cell Rep 17:236–242CrossRefGoogle Scholar
  44. Rao KS (1988) In Vitro meristem cloning of Eucalyptus tereticornis Sm. Plant Cell Rep 7:546–549CrossRefGoogle Scholar
  45. Rao RV, Shashikala S, Sreevani P, Kothiyal V, Sarma CR, Lal P (2002) Within tree variation in anatomical properties of some clones of Eucalyptus tereticornis Sm. Wood Sci Technol 36:271–285CrossRefGoogle Scholar
  46. Rossi F, Baraldi R, Facini O (1993) Photomorphogenic effects on In Vitro rooting of Prunus rootstock GF 655-2. Plant Cell Tissue Organ Cult 32:145–151CrossRefGoogle Scholar
  47. Rout GR, Mahato A, Senapati SK (2008) In Vitro clonal propagation of Nyctanthes arbortristis. Biol Plant 52:521–524CrossRefGoogle Scholar
  48. Saritha KV, Naidu CV (2008) Direct shoot regeneration from leaf explants of Spilanthes acmella. Biol Plant 52:334–338CrossRefGoogle Scholar
  49. Sharma AK, Raghuram N, Chandhok MR, Das R, Sopory SK (1999) Investigations on the nature of the phytochrome induced transmitters for the regulation of nitrate reductase in etiolated leaves of maize. J Exp Bot 45:485–490Google Scholar
  50. Sharma SK, Ramamurthy V (2000) Micropropagation of 4-yr-old elite Eucalyptus tereticornis trees. Plant Cell Rep 19:511–518CrossRefGoogle Scholar
  51. Sugimura Y, Kadotani N, Ueda Y, Shima K, Kitajima S, Furusawa T, Ikegami M (2005) Transgenic patchouli plants produced by Agrobacterium-mediated transformation. Plant Cell Tissue Organ Cult 82:251–257CrossRefGoogle Scholar
  52. Tanaka M, Takamura T, Watanabe H, Endo M, Yanagi T, Okamoto K (1998) In Vitro growth of Cymbidium plantlets cultured under super bright red and blue light-emitting diodes (LEDs). J Hortic Sci Biotechnol 73:39–44Google Scholar
  53. Thomas TD (2007) High-frequency, direct bulblet induction from rhizome explants of Curculigo orchioides Gaertn., an endangered medicinal herb. In Vitro Cell Dev Biol Plant 43:442–448CrossRefGoogle Scholar
  54. Trivedi P, Pandey A (2007) Biological hardening of micropropagated Picrorhiza kurrooa Royel ex Benth. an endangered species of medical importance. World J Microbiol Biotechnol 23:877–878CrossRefGoogle Scholar
  55. Vengadesan G, Pijut PM (2009) In Vitro propagation of northern red oak (Quercus rubra L.). In Vitro Cell Dev Biol Plant 45:474–482CrossRefGoogle Scholar
  56. Wardipura R, Salahhuddin S, Baharsah JS, Harrans (1986) The effect of rooting medium, partial leaf area removal and plant growth regulators on the growth of cuttings of cocoa (Theobroma cocoa L.). Forum-Pascasarjana 1:1–12Google Scholar

Copyright information

© The Society for In Vitro Biology 2012

Authors and Affiliations

  • Diwakar Aggarwal
    • 1
  • Anil Kumar
    • 1
  • Jagriti Sharma
    • 1
  • M. Sudhakara Reddy
    • 1
  1. 1.TIFAC-Center of Relevance and Excellence in Agro and Industrial Biotechnology, Department of Biotechnology & Environmental SciencesThapar UniversityPatialaIndia

Personalised recommendations