Skip to main content
SpringerLink
Log in

Auxin stability and accumulation during in vitro shoot morphogenesis influences subsequent root induction and development in Eucalyptus grandis

  • Original paper
  • Published:
Plant Growth Regulation Aims and scope Submit manuscript

Abstract

Recent results showed that after 16 months in the field, micropropagated eucalyptus plants have an inferior root system to cuttings. Such differences may be due to the plant growth regulators supplied during the culture stages of standard protocols, which are targeted at optimising plantlet yields and not root quality. This study investigated such a proposal, focusing on auxins in an easy-to-root clone. Initial results showed that the auxin provided in the standard protocol (NAA for multiplication and IBA for elongation) enabled 100% rooting in auxin-free medium, where rooting was faster than on IBA-rooting media. When auxin supply was omitted from multiplication and restricted to NAA or IAA during elongation, rooting in an auxin-free medium was reduced to 68 and 31%, respectively, reflecting the stabilities of these auxins in plant tissues. Additionally, 15% of shoots from the NAA-medium and 65% from the IAA-medium produced roots with altered graviperception. GC–MS analysis of these shoots revealed a relationship between free IAA-availability and altered graviperception. This was further tested by adding the IAA-specific transport inhibitor 2,3,5-triiodobenzoic acid to rooting media with IBA, IAA or NAA, which resulted in 100, 70.9 and 20.6% rooting, respectively. At least 40% of the sampled root tips had atypical starch grain deposition and abnormal graviperception. It is proposed that, at least in this clone, while IBA and NAA can be used for in vitro root induction, IAA is necessary for development of graviresponse.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

AG:

Altered graviperception

GC–MS:

Gas chromatography-mass spectrometry

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

NAA:

α-Naphthalene acetic acid

TIBA:

2,3,5-triiodobenzoic acid

PGR:

Plant growth regulator

References

  • Bell DT, van der Moezel PG, Bennett IJ, McComb JA, Wilkins CF, Marshall SCB, Morgan AL (1993) Comparisons of growth of Eucalyptus camaldulensis from seeds and tissue culture: root, shoot and leaf morphology of 9-month-old plants grown in deep sand and sand over clay. For Ecol Manag 57:125–139

    Article  Google Scholar 

  • Bennett MJ, Marchant A, Green HG, May ST, Ward SP, Millner PA, Walker AR, Schulz B, Feldmann KA (1996) Arabidopsis AUX1 gene: a permease-like regulator of root gravitropism. Science 273:948–950

    Article  PubMed  CAS  Google Scholar 

  • Blakesley D (1994) Auxin metabolism and adventitious root formation. In: Davis TD, Haissig BE (eds) The biology of adventitious root formation. Plenum Press, New York, pp 143–153

    Google Scholar 

  • Centeno ML, Fernández B, Feito I, Rodríguez A (1999) Uptake, distribution, and metabolism of 1-naphthaleneacetic acid and indole-3-acetic acid during callus initiation from Actinidia deliciosa tissues. J Plant Growth Regul 18:81–88

    Article  PubMed  CAS  Google Scholar 

  • Chen R, Rosen E, Masson PH (1999) Gravitropism in higher plants. Plant Physiol 120:343–350

    Article  PubMed  CAS  Google Scholar 

  • 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 87:207–214

    Article  CAS  Google Scholar 

  • de 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, India, pp 303–333

    Google Scholar 

  • De Klerk G-J, Van der Krieken W, De Jong J (1999) The formation of adventitious roots: new concepts, new possibilities. In Vitro Cell Dev Biol Plant 35:189–199

    Article  Google Scholar 

  • Delbarre A, Muller P, Imhoff V, Guern J (1996) Comparison of mechanisms controlling uptake and accumulation of 2,4-dichlorophenoxyacetic acid, naphthalene-1-acetic acid, and indole-3-acetic acid in suspension-cultured tobacco cells. Planta 198:532–541

    Article  CAS  Google Scholar 

  • Denison NP, Kietzka JE (1993) The development and utilisation of vegetative propagation in Mondi for commercial afforestation programmes. S Afr For J 165:47–54

    Google Scholar 

  • Eldridge K, Davidson J, Harwood C, van Wyk G (1994) Eucalypt domestication and breeding. Claredon Press, London

    Google Scholar 

  • Epstein E, Ludwig-Müller J (1993) Indole-3-butyric acid in plants: occurrence, synthesis, metabolism and transport. Physiol Plant 88:382–389

    Article  CAS  Google Scholar 

  • Evans ML (1991) Gravitropism: interaction of sensitivity modulation and effector redistribution. Plant Physiol 95:1–5

    Article  PubMed  CAS  Google Scholar 

  • Fett-Neto AG, Pett JP, Goulart LWV, Pasquali G, Termignoni RR, Ferreira AG (2001) Distinct effects of auxin and light on adventitious root development in Eucalyptus saligna and Eucalyptus globulus. Tree Physiol 21:457–464

    PubMed  CAS  Google Scholar 

  • Fogaça CM, Fett-Neto AG (2005) Role of auxin and its modulators in the adventitious rooting of Eucalyptus species differing in recalcitrance. Plant Growth Regul 45:1–10

    Article  Google Scholar 

  • George EF, Hall MA, De Klerk G-J (eds) (2008) Plant propagation by tissue culture, 3rd edn. Springer, The Netherlands, pp 175–204

    Google Scholar 

  • Goren R, Bukovac MJ (1973) Mechanism of naphthaleneacetic acid conjugation. Plant Physiol 51:907–913

    Article  PubMed  CAS  Google Scholar 

  • Hammer Ø, Harper DAT, Ryan PD (2001) PAST: paleontological statistics software package for education and data analysis. Palaeontologia Electron 4:9

    Google Scholar 

  • Jones NB, Van Staden J (1997) Micropropagation of Eucalyptus. In: Bajaj YPS (ed) Biotechnology in agriculture and forestry (vol 39), high-tech and micropropagation. Springer, Berlin, pp 286–329

    Google Scholar 

  • Kiss JZ (2000) Mechanisms of the early phases of plant gravitropism. Crit Rev Plant Sci 19:551–573

    Article  PubMed  CAS  Google Scholar 

  • Ludwig-Müller J (2000) Indole-3-butyric acid in plant growth and development. Plant Growth Regul 32:219–230

    Article  Google Scholar 

  • Ludwig-Müller J, Epstein E (1991) Occurance and in vivo biosynthesis of indole-3-butyric acid in corn (Zea mays L.). Plant Physiol 97:765–770

    Article  PubMed  Google Scholar 

  • Mokotedi MEO, Watt MP, Pammenter NW, Blakeway FC (2000) In vitro rooting and subsequent survival of two clones of a cold-tolerant Eucalyptus grandis × E. nitens hybrid. Hortscience 35:1163–1165

    Google Scholar 

  • Mokotedi MEO, Pammenter NW, Watt MP, Blakeway FC (2003) Physiological characteristics of nine-month-old Eucalyptus grandis × nitens plants produced through micropropagation and macropropagation. Acta Hortic 616:407–411

    Google Scholar 

  • Mokotedi MEO, Watt MP, Pammenter NW (2009) The influence of vegetative propagation on root functioning in Eucalyptus grandis × nitens. Acta Hortic 812:395–402

    Google Scholar 

  • Mokotedi MEO, Watt MP, Pammenter NW (2010) Analysis of differences in field performance of vegetativley and seed-propagated Eucalyptus varieties II: vertical uprooting resistance. South For 72:31–36

    Google Scholar 

  • Morita MT, Tasaka M (2004) Gravity sensing and signalling. Curr Opin Plant Biol 7:712–718

    Article  PubMed  CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nordström A-C, Jacobs FA, Eliasson L (1991) Effect of exogenous indole-3-acetic acid and indole-3-butyric acid on internal levels of the respective auxins and their conjugation with aspartic acid during adventitious root formation in pea cuttings. Plant Physiol 96:856–861

    Article  PubMed  Google Scholar 

  • Palme K, Gälweiler L (1999) PIN-pointing the molecular basis of auxin transport. Curr Opin Plant Biol 2:375–381

    Article  PubMed  CAS  Google Scholar 

  • Poupart J, Waddell CS (2000) The rib1 mutant is resistant to indole-3-butyric acid, an endogenous auxin in Arabidopsis. Plant Physiol 124:1739–1751

    Article  PubMed  CAS  Google Scholar 

  • Rashotte AM, Brady SR, Reed RC, Ante SJ, Muday GK (2000) Basipetal auxin transport is required for gravitropism in roots of Arabidopsis. Plant Physiol 122:481–490

    Article  PubMed  CAS  Google Scholar 

  • Schwambach J, Ruedell CM, de Almeida MR, Penchel RM, de 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 36:261–271

    Article  Google Scholar 

  • Smulders MJM, Van de Ven ETWM, Croes AF, Wullems GJ (1990) Metabolism of 1-naphthaleneacetic acid in explants of tobacco: evidence for release of free hormone from conjugates. J Plant Growth Regul 9:27–34

    Article  CAS  Google Scholar 

  • Spurr AR (1969) A low viscosity epoxy resin embedding medium for electron microscopy. J Ultrastruct Res 26:31–43

    Article  PubMed  CAS  Google Scholar 

  • Stange L (1985) Effects of TIBA on meristematic activity and starch metabolism in Riella helicophylla (Bory et Mont.) Mont. Biol Plant 27:221–225

    Article  CAS  Google Scholar 

  • Vieten A, Sauer M, Brewer PB, Friml J (2007) Molecular and cellular aspects of auxin-transport-mediated development. Trends Plant Sci 12:160–168

    Article  PubMed  CAS  Google Scholar 

  • Watt MP, Blakeway FC, Mokotedi MEO, Jain SM (2003) Micropropagation of Eucalyptus. In: Jain SM, Ishii K (eds) Micropropagation of woody trees and fruits. Kluwer Academic Publishers, Netherlands, pp 217–244

    Chapter  Google Scholar 

  • Wiesman Z, Riov J, Epstein E (1989) Characterization and rooting ability of indole-3-butyric acid conjugates formed during rooting of mung bean cuttings. Plant Physiol 91:1080–1084

    Article  PubMed  CAS  Google Scholar 

  • Woodward AW, Bartel B (2005) Auxin: regulation, action and interaction. Ann Bot 95:707–735

    Article  PubMed  CAS  Google Scholar 

  • Yamamoto M, Yamamoto KT (1998) Differential effects of 1-naphthaleneacetic acid, indole-3-acetic acid and 2,4-dichlorophenoxyacetic acid on the gravitropic response of roots in an auxin-resistant mutant of Arabidopsis, aux1. Plant Cell Physiol 39:660–664

    PubMed  CAS  Google Scholar 

  • Yasodha R, Sumathi R, Gurumurthi K (2004) Micropropagation for quality propagule production in plantation forestry. Indian J Biotechnol 3:159–170

    Google Scholar 

Download references

Acknowledgments

The National Research Fund, Universities of KwaZulu-Natal and Witwatersrand, and Mondi Business are gratefully acknowledged for continued funding for this research. M. Nakhooda also thanks the UKZN- LEAP programme for its support and Dr M. van der Merwe for expert assistance with the PGR assays.

Author information

Authors and Affiliations

  1. School of Biological and Conservation Sciences, University of KwaZulu-Natal, Durban, South Africa

    Muhammad Nakhooda & M. Paula Watt

  2. School of Animal, Plant and Environmental Sciences, University of the Witwatersrand, Johannesburg, South Africa

    David Mycock

Authors
  1. Muhammad Nakhooda
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Paula Watt
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. David Mycock
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Muhammad Nakhooda.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Nakhooda, M., Watt, M.P. & Mycock, D. Auxin stability and accumulation during in vitro shoot morphogenesis influences subsequent root induction and development in Eucalyptus grandis . Plant Growth Regul 65, 263–271 (2011). https://doi.org/10.1007/s10725-011-9597-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10725-011-9597-7

Keywords

Search

Navigation