New Forests

, Volume 46, Issue 3, pp 357–371 | Cite as

Micropropagation of Eucalyptus camaldulensis for the production of rejuvenated stock plants for microcuttings propagation and genetic fidelity assessment

  • K. Shanthi
  • V. K. W. Bachpai
  • S. Anisha
  • M. Ganesan
  • R. G. Anithaa
  • V. Subashini
  • M. Chakravarthi
  • V. Sivakumar
  • R. Yasodha
Article
First Online:
Received:
Accepted:

Abstract

Adventitious rooting, a key step in clonal propagation is affected by tree maturation. Micropropagation followed by microcuttings propagation has the potential to rejuvenate the clones thereby enhancing the rooting potential and minimize intra clonal variation. In this study, 33 superior performing Eucalyptus camaldulensis clones were propagated by rooting of stem cuttings (SCs) and micropropagation. Micropropagated plantlets were used as stock plants for microcutting propagation. Rooting of SCs and micropropagation was carried out with the coppice shoot cuttings and axillary buds respectively, obtained from approximately fourteen year old trees that had undergone one vegetative propagation cycle. The adventitious rooting recorded was significantly higher in micropropagation (24.8–100 %) and microcuttings (43–95 %) than SCs method (9.3–75.5 %). Studies on ontogeny of adventitious rooting showed the emergence of root primordium from the phloem region and root initials were noticed within 5–9 days after auxin treatment. Further, molecular marker analysis showed genetic uniformity except for two ramets, detected using inter simple sequence repeats (ISSR) markers and suitable corrective measures were taken to avoid entry of such plantlets for mass multiplication. This study demonstrates the importance of integration of micropropagation and microcuttings production for rejuvenation and mass multiplication. Although current rejuvenation and root induction treatments favored adventitious rooting, the basic mechanisms involved in rejuvenation and adventitious rooting need to be explored for hassle free industrial rooting process, consequently cost effective propagation.

Keywords

Adventitious rooting Stem cuttings Micropropagation Microcuttings Ontogeny of roots Genetic fidelity 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

This works forms the part of the Indian Council of Forestry Research and Education (ICFRE) funded project. Authors S. V. and C. M. acknowledge the junior research fellowship from ICFRE, Dehradun, India.

References

  1. Amissah JN, Paolillo DJ Jr, Bassuk N (2008) Adventitious root formation in stem cuttings of Quercus bicolor × Quercus macrocarpa and its relationship to stem anatomy. J Am Soc Hortic Sci 133(4):479–486Google Scholar
  2. Assis TF, Fett-Neto AG, Alfenas AC (2004) Current techniques and prospects for the clonal propagation of hardwoods with emphasis on Eucalyptus. In: Walter C, Carson M (eds) Plantation forest biotechnology for the 21st century. Research Signpost, Trivandrum, pp 303–333Google Scholar
  3. Ballester A, San José MC, Vidal N, Fernandez-Lorenzo JL, Vieitez AM (1999) Anatomical and biochemical events during in vitro rooting of microcuttings from juvenile and mature phases of chestnut. Ann Bot 83(6):619–629CrossRefGoogle Scholar
  4. Baltierra XC, Montenegro G, De Garcia E (2004) Ontogeny of in vitro rooting processes in Eucalyptus globulus. In Vitro Cell Dev Biol Plant 40(5):499–503CrossRefGoogle Scholar
  5. Brondani GE, De Wit Ondas HW, Baccarin FJB, Gonçalves AN, De Almeida M (2012) Micropropagation of Eucalyptus benthamii to form a clonal micro-garden. In Vitro Cell Dev Biol Plant 48(5):478–487CrossRefGoogle Scholar
  6. Cheng B, Peterson CM, Mitchell RJ (1992) The role of sucrose, auxin and explant source on in vitro rooting of seedling explants of Eucalyptus sideroxylon. Plant Sci 87(2):207–214CrossRefGoogle Scholar
  7. Clarke PJ, Lawes MJ, Midgley JJ, Lamont BB, Ojeda F, Burrows GE, Enright NJ, Knox KJE (2013) Resprouting as a key functional trait: how buds, protection and resources drive persistence after fire. New Phytol 197(1):19–35CrossRefPubMedGoogle Scholar
  8. Goodger JQD, Woodrow IE (2010) The influence of micropropagation on growth and coppicing ability of Eucalyptus polybractea. Tree Physiol 30(2):285–296CrossRefPubMedGoogle Scholar
  9. Iglesias Trabado G, Wilstermann D (2008) Eucalyptus universalis. Global cultivated eucalypt forests map 2008 Ver 1.0.1. In: GIT Forestry consulting’s EUCALYPTOLOGICS, information resources on Eucalyptus cultivation worldwide. Retrieved from http://www.git-forestry.com/ accessed 29 March 2009
  10. Le Roux JJ, Staden JV (1991) Micropropagation and tissue culture of Eucalyptus—a review. Tree Physiol 9(4):435–477CrossRefPubMedGoogle Scholar
  11. Lee SJ (2001) Selection of parents for the sitka spruce breeding population in Britain and the strategy for the next breeding cycle. Forestry 74(2):129–143CrossRefGoogle Scholar
  12. Majada J, Martínez-Alonso C, Feito I, Kidelman A, Aranda I, Alía R (2011) Mini-cuttings: an effective technique for the propagation of Pinus pinaster Ait. New For 41(3):399–412CrossRefGoogle Scholar
  13. Makouanzi G, Bouvet JM, Denis M, Saya A, Mankessi F, Vigneron P (2014) Assessing the additive and dominance genetic effects of vegetative propagation ability in Eucalyptus—influence of modeling on genetic gain. Tree Genet Genomes 5(10):1243–1256CrossRefGoogle Scholar
  14. Mankessi F, Saya A, Toto M, Monteuuis O (2011) Cloning field growing Eucalyptus urophylla × Eucalyptus grandis by rooted cuttings: age, within-shoot position and season effects. Propag Ornam Plants 11(1):3–9Google Scholar
  15. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15(3):473–497CrossRefGoogle Scholar
  16. Negishi N, Nakahama K, Urata N, Kojima M, Sakakibara H, Kawaoka A (2014) Hormone level analysis on adventitious root formation in Eucalyptus globulus. New For 45(4):577–587CrossRefGoogle Scholar
  17. Nourissier S, Monteuuis O (2008) In vitro rooting of two Eucalyptus urophylla × Eucalyptus grandis mature clones. In Vitro Cell Dev Biol Plant 44(4):263–272CrossRefGoogle Scholar
  18. Oliveira LS, Xavier A, Dias PC, Takahashi EK, Paiva HN (2012) Rooting of mini-cuttings and micro-cuttings of Eucalyptus urophylla × E. globulus and Eucalyptus grandis × E. globulus. Sci For 40(96):507–516Google Scholar
  19. Pathak H, Dhawan V (2012) ISSR assay for ascertaining genetic fidelity of micropropagated plants of apple rootstock Merton 793. In Vitro Cell Dev Biol Plant 48:137–143CrossRefGoogle Scholar
  20. Phulwaria M, Patel AK, Rathore JS, Ram K, Shekhawat NS (2014) An improved micropropagation and assessment of genetic stability of micropropagated Salvadora oleoides using RAPD and ISSR markers. Acta Physiol Plant 36(5):1115–1122CrossRefGoogle Scholar
  21. Pijut PM, Woeste KE, Michler CH (2011) Promotion of adventitious root formation of difficult-to-root hardwood tree species. Hortic Rev 38:213–251Google Scholar
  22. Pinto G, Araújo C, Santos C, Neves L (2013) Plant regeneration by somatic embryogenesis in Eucalyptus spp. Current status and future perspectives. South For J For Sci 75(2):59–69Google Scholar
  23. Rezende GDSP, Resende MDV, Assis TF (2014) A roadmap to Eucalyptus breeding for clonal forestry. In: Fenning T (ed) Challenges and opportunities for the world’s forests in the 21st century. Springer, Dordrecht, pp 393–424CrossRefGoogle Scholar
  24. Rockwood DL, Rudie AW, Ralph SA, Zhu JY, Winandy JE (2008) Energy product options for Eucalyptus species grown as short rotation woody crops. Int J Mol Sci 9(8):1361–1378CrossRefPubMedCentralPubMedGoogle Scholar
  25. Ruedell CM, De Almeida MR, Schwambach J, Posenato C, Fett-Neto AG (2013) Pre and post-severance effects of light quality on carbohydrate dynamics and microcutting adventitious rooting of two Eucalyptus species of contrasting recalcitrance. Plant Growth Regul 69(3):235–245CrossRefGoogle Scholar
  26. San José MC, Romero L, Janeiro LV (2012) Effect of indole-3-butyric acid on root formation in Alnus glutinosa microcuttings. Silva Fenn 46(5):643–654CrossRefGoogle Scholar
  27. Saya RA, Mankessi F, Toto M, Marien JN, Monteuuis O (2008) Advances in mass clonal propagation of Eucalyptus urophylla × E. grandis in Congo. Bois For Trop 297(3):15–25Google Scholar
  28. Schwambach J, Ruedell CM, Almeida MR, Penchel RM, Araújo EF, Fett-Neto AG (2008) Adventitious rooting of Eucalyptus globulus × maidennii mini-cuttings derived from mini-stumps grown in sand bed and intermittent flooding trays: a comparative study. New For 36(3):261–271CrossRefGoogle Scholar
  29. Steinitz B, Barr N, Tabib Y, Vaknin Y, Bernstein N (2010) Control of in vitro rooting and plant development in Corymbia maculata by silver nitrate, silver thiosulfate and thiosulfate ion. Plant Cell Rep 29(11):1315–1323CrossRefPubMedGoogle Scholar
  30. Titon M, Xavier A, Otoni WC (2006) Clonal propagation of Eucalyptus grandis using the mini-cutting and micro-cutting techniques. Sci For 71:109–117Google Scholar
  31. Tripathi S, Mathish N, Gurumurthi K (2006) Use of genetic markers in the management of micropropagated Eucalyptus germplasm. New For 31(3):361–372CrossRefGoogle Scholar
  32. Trueman SJ (2006) Clonal propagation and storage of subtropical pines in Queensland, Australia. South Afr For J 208(1):49–52Google Scholar
  33. Trueman SJ, McMahon TV, Bristow M (2013) Production of Eucalyptus cloeziana cuttings in response to stock plant temperature. J Trop For Sci 25(1):60–69Google Scholar
  34. Varghese M, Harwood CE, Hegde R, Ravi N (2008) Evaluation of provenances of Eucalyptus camaldulensis and clones of E. camaldulensis and E. tereticornis at contrasting sites in southern India. Silva Genet 57(3):136–141Google Scholar
  35. Verstraeten I, Beeckman T, Geelen D (2013) Adventitious root induction in Arabidopsis thaliana as a model for in vitro root organogenesis. Methods Mol Biol 959:159–175CrossRefPubMedGoogle Scholar
  36. Vivekanandan K, Gurumurthi K, Jayaraj RSC (1997) Clonal multiplication of Eucalyptus—Field Document: UNDP/FAO Regional project on improved productivity of man-made forests through application of technological advances in tree breeding and propagation (RAS/91/004—FORTIP). FAO, Philippines, Los BanosGoogle Scholar
  37. Watt MP (2014) Genotypic-unspecific protocols for the commercial micropropagation of Eucalyptus grandis × nitens and E. grandis × urophylla. Turk J Agric For 38:125–133CrossRefGoogle Scholar
  38. Wendling I, Brondani GE, Dutra LF, Hansel FA (2010) Mini-cuttings technique: a new ex vitro method for clonal propagation of sweetgum. New For 39:343–353CrossRefGoogle Scholar
  39. Wendling I, Trueman SJ, Xavier A (2014a) Maturation and related aspects in clonal forestry-part I: concepts, regulation and consequences of phase change. New For 45(4):449–471CrossRefGoogle Scholar
  40. Wendling I, Trueman SJ, Xavier A (2014b) Maturation and related aspects in clonal forestry-part II: reinvigoration, rejuvenation and juvenility maintenance. New For 45(4):473–486CrossRefGoogle Scholar
  41. Whittock SP, Apiolaza LA, Kelly CM, Potts BM (2003) Genetic control of coppice and lignotuber development in Eucalyptus globulus. Aust J Bot 51:57–67CrossRefGoogle Scholar
  42. Yasodha R, Sumathi R, Gurumurthi K (1997) Micropropagation of difficult to propagate clones of eucalypts. IUFRO conference on silviculture and improvement of Eucalyptus. Embrapa, Colombo, pp 123–128Google Scholar
  43. Yasodha R, Sumathi R, Gurumurthi K (2002) Micropropagation of a putative hybrid E. torrelliana × E. citriodora. In: Bagchi SK, Mohan V, Siddappa (eds) Recent eucalypt research in India. ICFRE, Dehradun, pp 129–135Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • K. Shanthi
    • 1
  • V. K. W. Bachpai
    • 1
  • S. Anisha
    • 1
  • M. Ganesan
    • 1
  • R. G. Anithaa
    • 1
  • V. Subashini
    • 1
  • M. Chakravarthi
    • 1
  • V. Sivakumar
    • 1
  • R. Yasodha
    • 1
  1. 1.Institute of Forest Genetics and Tree BreedingCoimbatoreIndia

Personalised recommendations