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Plant Cell, Tissue and Organ Culture

, Volume 90, Issue 1, pp 15–23 | Cite as

Optimizing the micropropagation protocol for the endangered Aloe polyphylla: can meta-topolin and its derivatives serve as replacement for benzyladenine and zeatin?

  • Michael W. Bairu
  • Wendy A. Stirk
  • Karel Dolezal
  • Johannes Van StadenEmail author
Original Paper

Abstract

Benzyladenine (BA) is the most widely used cytokinin in the micropropagation industry due to its effectiveness and affordability. It, however, has disadvantages such as genetic alteration and abnormal growth in some plants. Naturally occurring zeatin on the other hand is not as widely used as BA and is far more expensive. The use of meta-topolin and its derivatives as alternatives to BA and zeatin, both of which frequently have negative effects in tissue culture was investigated. In vitro grown Aloe polyphylla (an endangered medicinal and ornamental aloe) were cultured on full strength Murashige and Skoog basal medium with different concentrations of cytokinins and solidified with 1% Bacteriological Agar (Oxoid No. 1). mT was the preferred cytokinin both in terms of multiplication rate and rooting. The optimum concentration that induced regeneration and rooting is 5.0 μM. The problem of hyperhydricity was totally controlled. Plants rooted spontaneously in multiplication medium, thus avoiding the extra rooting step of the protocol. More than 91% of the plants transferred to ex vitro conditions were successfully acclimatized.

Keywords

Aromatic cytokinins Conservation Hyperhydricity Metabolites Multiplication rate Plant growth regulators Root and shoot growth 

Abbreviations

[9G]BA

[6-benzylamino-9- β-d-glucopyranosylpurine]

BA

[6-benzylaminopurine]

IBA

indole butyric acid

MS

Murashige and Skoog (1962) medium

MemT

[6-(3-methoxybenzylamino)purine]

MemTR

[6-(3-methoxybenzylamino)-9-β-d-ribofuranosylpurine]

mT

[6-(3-hydroxybenzylamino)purine]

mTR

[6-(3-hydroxybenzylamino)-9-β-d-ribofuranosylpurine]

oT

[6-(2-hydroxybenzylamino)purine]

PGR

plant growth regulators

pT

[6-(4-hydroxybenzylamino)purine]

Notes

Acknowledgement

We acknowledge the financial support of the National Research Foundation (NRF), Pretoria. This work was also supported by the Grant Agency of the Czech Republic (GA 522/06/0108).

References

  1. Abrie AL, Van Staden J (2001) Micropropagation of the endangered Aloe polyphylla. Plant Growth Regul 33:19–23CrossRefGoogle Scholar
  2. Aubrey A (2002) Aloe polyphylla. Available online at http://www.plantzafrica.com/plantab/aloepoly.htm Cited 11 Sept 2005
  3. Baroja-Fernandez E, Aguirreolea J, Martínková H, Hanus J, Strnad M (2002) Aromatic cytokinins in micropropagated potato plants. Plant Physiol Biochem 40:217–224CrossRefGoogle Scholar
  4. Chukwujechwu JC, Fennell CW, van Staden J (2002) Optimisation of the tissue culture protocol for the endangered Aloe polyphylla. S Afr J Bot 68:424–429Google Scholar
  5. Horgan R, Hewett EW, Horgan JM, Purse JG, Wareing PF (1975) A new cytokinin from Populus × robusta. Phytochemistry 19:1005–1008CrossRefGoogle Scholar
  6. Iwamura H, Fujiita T, Koyama S, Koshimizu K, Kumazawa Z (1980) Quantitative structure-activity relationship of cytokinin-active adenine and urea derivatives. Phytochemistry 19:1309–1319Google Scholar
  7. Jones LH, Martinková H, Strnad M, Hanke DE (1996) Occurrence of aromatic cytokinins in oil palm (Elaeis guineensis Jacq.). J Plant Growth Regul 15:39–49CrossRefGoogle Scholar
  8. Kamínek M, Paces V, Corse J, Challice JS (1979) Effect of stereospecific hydroxylation of N6-(Δ2-Isopentenyl)adenosine on cytokinin activity. Planta 145:239–243CrossRefGoogle Scholar
  9. Kamínek M, Vanék T, Kalendova A, Pilar J (1987a) Effect of two cytokinins on production of stem cuttings by stock plants of Euphorbia pulcherrima Wild. and Gerbera jamesonii Hook. Sci Hortic 33:281–289CrossRefGoogle Scholar
  10. Kamínek M, Vanék T, Motyka V (1987b) Cytokinin activities of N6-benzyladenosine derivatives hydroxylated on the side-chain phenyl ring. J Plant Growth Regul 6:113–120CrossRefGoogle Scholar
  11. Krikorian AD (1995) Hormones in tissue culture and micropropagation. In: Davies PJ (eds) Plant hormones: physiology, biochemistry and molecular biology. Academic Publishers, The Netherlands, pp 774–796Google Scholar
  12. Kubalakova M, Strnad M (1992) The effect of aromatic cytokinins (populins) on micropropagation and regeneration in vitro. Biol Plant 34:578–579Google Scholar
  13. Leshem B, Sachs T (1985) ‘Vitrified’ Dianthus- teratomata in vitro due to growth factor imbalance. Ann Bot 56:613–617Google Scholar
  14. Leshem B, Shaley DP, Izhar S (1988) Cytokinin as an inducer of hyperhydricity in melon. Ann Bot 61:255–260 Google Scholar
  15. Miller CO (1965) Evidence for the natural occurrence of zeatin and derivatives: compounds from maize which promote cell division. Proc Natl Acad Sci USA 54:1052–1058PubMedCrossRefGoogle Scholar
  16. Mok MC, Martin RC, Dobrev PI, Vancovà R, Ho PS, Yonekura-Sakakibara K, Sakakibara H, Mok DWS (2005) Topolins and hydroxylated thidiazuron derivatives are substrates of cytokinin o-glucosyltransferase with position specificity related to receptor recognition. Plant Physiol 137:1057–1066PubMedCrossRefGoogle Scholar
  17. Murashige T, Skoog F (1962) A revised medium for a rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  18. Strnad M (1997) The aromatic cytokinins. Physiol Plant 101:674–688CrossRefGoogle Scholar
  19. Strnad M, Hanuš J, Vanĕk T, Kamínek M, Ballantine JA, Fussell B, Hanke DE (1997) Meta-topolin, a highly active aromatic cytokinin from poplar leaves (Populus × canadensis Moench, cv. Robusta). Phytochemistry 45:213–218CrossRefGoogle Scholar
  20. Tarkowská D, Doležal K, Tarkowski P, Åstot C, Holub J, Fuksová K, Schmülling T, Sandberg G, Strnad M (2003) Identification of new aromatic cytokinins in Arabidopsis thaliana and Populus × canadensis leaves by LC-(+)ESI-MS and capillary liquid chromatography frit-fast atom bombardment mass spectrometry. Physiol Plant 117(4):579–590PubMedCrossRefGoogle Scholar
  21. Teramoto H, Momotani E, Tsuji H (1993) Benzyladenine-induced changes in the translatable mRNA population in excised cucumber cotyledons. Physiol Plant 87:584–591CrossRefGoogle Scholar
  22. Van Staden J, Drewes FE (1991) The biological activity of cytokinin derivatives in soybean callus bioassay. Plant Growth Regul 10:109–115CrossRefGoogle Scholar
  23. Werbrouck SPO, Strnad M, Van Onckelen HA, Debergh PC (1996) Meta-topolin, an alternative to benzyladenine in tissue culture. Physiol Plant 98:291–297CrossRefGoogle Scholar
  24. Werbrouck SPO, Van der Jeugt B, Dewitte W, Prinsen E, Van Onckelen HA, Debergh PC (1995) The metabolism of benzyladenine in Spathiphyllum floribundum schott ‘petite’ in relation to acclimatization problems. Plant Cell Rep 14:662–665CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media B.V. 2007

Authors and Affiliations

  • Michael W. Bairu
    • 1
  • Wendy A. Stirk
    • 1
  • Karel Dolezal
    • 2
  • Johannes Van Staden
    • 1
    Email author
  1. 1.Research Centre for Plant Growth and Development, School of Biological and Conservation SciencesUniversity of KwaZulu-Natal Pietermaritzburg ScottsvilleSouth Africa
  2. 2.Laboratory of Growth RegulatorsPalacky University and Institute of Experimental Botany AS CROlomouc-HoliceCzech Republic

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