Advertisement

Micropropagation of Wild Service Tree (Sorbus torminalis [L.] Crantz): The Regulative Role of Different Aromatic Cytokinins During Organogenesis

  • Jana Malá
  • Pavlína Máchová
  • Helena Cvrčková
  • Michal Karady
  • Ondřej Novák
  • Jaromír Mikulík
  • Eva Hauserová
  • Jarmila Greplová
  • Miroslav Strnad
  • Karel DoležalEmail author
Article

Abstract

The influences of three different aromatic cytokinin derivatives [6-benzylaminopurine, meta-topolin, and 6-(3-methoxybenzylamino)purine-9-ß-D-ribofuranoside (MeOBAPR)] on in vitro multiplication and rhizogenesis of the wild service tree (Sorbus torminalis [L.] Crantz) were compared. The highest micropropagation rate (24 new shoots per explant after 3 months of cultivation) was achieved on media containing BAP. On the other hand, the best rooting microcuttings were those multiplied on a medium containing MeoBAPR. To compare these results with the levels of endogenous cytokinins in multiplied explants, a newly developed UPLC-ESI(+)-MS/MS method was used to determine levels of 50 cytokinin metabolites in explants cultivated 12 weeks on media supplemented by BAP and of the two other aromatic cytokinin analogs used. Several significant differences among the levels of endogenous cytokinins, extracted from the explants, were found. The concentration of BAP9G, an important metabolite suspected to be responsible for inhibition of rooting and acclimatization problems of newly formed plantlets, was found to be the highest in microcuttings grown on media supplemented with BAP. This agrees well with the results of our rooting experiments; the lowest percentages of rooted plantlets 6 weeks after transferring shoots on rooting medium were present on explants multiplied on BAP. In contrast, BAP was still the most effective for the induction of bud formation on primary explants. Levels of the most active endogenous isoprenoid cytokinins, tZ, tZR, and iPR, as well as O-glucosides were also suppressed in explants grown on BAP compared with those of explants treated with other cytokinin derivatives. This may be the result of a very high BAP uptake into the explants grown on this cytokinin. On the other hand, endogenous concentrations of cis-zeatin derivatives as well as dihydrozeatin derivatives were not affected. Differences in the production of another plant hormone, ethylene, that plays an important role in controlling organogenesis in tissue culture, were also observed among S. torminalis plantlets grown in vitro on media containing different cytokinins tested. The highest ethylene levels were detected in the vessels containing media supplemented with mT. They were two to four times higher compared with the production by the S. torminalis explants cultivated on other media used. Finally, the levels of free IAA were also determined in the explants. S. torminalis plantlets grown on media containing BAP contained the lowest level of auxin, which is again in good agreement with their loss of rooting capacity. The results found in this study about optimal plant hormone concentrations may be used to improve in vitro rooting efficiency of the wild service tree and possibly also of other plant species.

Keywords

Wild service tree Micropropagation Rooting Cytokinins Ethylene IAA 

Abbreviations

BAP

6-Benzylaminopurine

MeOBAP

6-(3-Methoxybenzylamino)purine

mT

6-(3-Hydroxybenzylamino)purine

oT

6-(2-Hydroxybenzylamino)purine

iP

N6-isopentenyladenine

cZ

cis-Zeatin

tZ

trans-Zeatin

DHZ

Dihydrozeatin

*R

9-β-d-ribofuranosyl derivative

*9G

9-β-d-glucopyranosyl derivative

*OG

O-β-d-glucopyranosyl derivative

*5′MP

5′-Monophosphate derivative

LC-MS

Liquid chromatography combined with mass spectrometry

UPLC-ESI(+)-MS/MS

Ultra-performance liquid chromatography combined with positive electrospray mass spectrometry

MS

Murashige-Skoog medium

IAA

Indole-3-acetic acid

IBA

Indole-3-butyric acid

NAA

α-Naphthylacetic acid

Notes

Acknowledgments

This work was supported by the Grant Agency of the Czech Republic (GA 206/07/0570), by the Ministry of Agriculture of the Czech Republic (MZE 0002070203), and the Czech Ministry of Education (MSM 6198959216, 1M06030). We thank Petra Amakorová for her skilled technical assistance.

References

  1. Altamura MM, Torrigiani P, Falasca G, Rossini P, Bagni N (1993) Morpho-functional gradients in superficial and deep tissues along tobacco stem: polyamine levels, biosynthesis and oxidation and organogenesis in vitro. J Plant Physiol 142:543–551Google Scholar
  2. Arigita L, Tames RS, Gonzales A (2003) 1-Methylcyclopropene and ethylene as regulators of in vitro organogenesis in kiwi explants. Plant Growth Regul 40:59–64CrossRefGoogle Scholar
  3. Bollmark M, Kubát B, Eliasson L (1988) Variation in endogenous cytokinin content during adventitious root formation in pea cuttings. J Plant Physiol 132:262–265Google Scholar
  4. Casson SA, Lindsey K (2003) Genes and signaling in root development. New Phytol 158:11–38Google Scholar
  5. Centeno ML, Rodríguez I, Feito I, Fernández B (1996) Relationship between endogenous auxin and cytokinin levels and morphogenic responses in Actinia deliciosa tissue cultures. Plant Cell Rep 16:58–62CrossRefGoogle Scholar
  6. Chalupa V (1992) Micropropagation of European mountain-ash (Sorbus aucuparia L.) and wild service tree (Sorbus torminalis (L.) Cr.). In: Bajaj YPS (ed) High-tech and micropropagation. II. Biotechnology in agriculture and forestry 18. Springer-Verlag, Berlin, pp 211–226Google Scholar
  7. Cvikrová M, Hrubcová M (1999) The role of phenolic substances in the processes of differentiation and morphogenesis. In: Strnad M, Peč P, Beck E (eds) Advances in regulation of plant development. Peres Publications, Prague, pp 213–220Google Scholar
  8. Davies FT Jr, Davis TD, Kestrer DE (1994) Commercial importance of adventitious rooting to horticulture. In: Davis TD, Haissig BE (eds) Biology of adventitious root formation. Plenum Press, New York, pp 53–59Google Scholar
  9. De Klerk GJ, Hanečáková J, Jasik J (2001) The role of cytokinins in rooting of stem slices cut from apple microcuttings. Plant Biosyst 135:79–84CrossRefGoogle Scholar
  10. Demesure B, Leguerroué B, Lucchi G, Prat D, Petit RJ (2000) Genetic variability of a scattered temperate forest tree: Sorbus torminalis L. (Crantz). Ann For Sci 57:63–71CrossRefGoogle Scholar
  11. Doležal K, Popa I, Kryštof V, Spíchal L, Fojtíková M, Holub J, Lenobel R, Schmülling T, Strnad M (2006) Preparation and biological activity of 6-benzylaminopurine derivatives in plants and human cancer cells. Bioorg Med Chem 14:875–884CrossRefPubMedGoogle Scholar
  12. Doležal K, Popa I, Hauserová E, Spíchal L, Chakrabarty K, Novák O, Kryštof V, Voller J, Holub J, Strnad M (2007) Preparation, biological activity and endogenous occurrence of N6-benzyladenosines. Bioorg Med Chem 15:3737–3747CrossRefPubMedGoogle Scholar
  13. Dujíčková M, Malá J, Chalupa V (1992) Vegetative reproduction of Sorbus torminalis L. Crantz and Sorbus domestica L. in vitro. Works FGMRI 77:27–48 (in Czech)Google Scholar
  14. Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42CrossRefGoogle Scholar
  15. Fišerová H, Kula E, Klemš M, Reinöhl V (2001) Phytohormones as indicators of the degree of damage in birch (Betula pendula). Biologia 56:405–409Google Scholar
  16. Fusseder A, Ziegler P (1988) Metabolism and compartmentation of dihydrozeatin exogenously supplied to photoautotrophic suspension-cultures of Chenopodium rubrum. Planta 173:104–109CrossRefGoogle Scholar
  17. Gonzalez A, Rodriguez R, Tames RS (1991) Ethylene and in vitro rooting of hazelnut (Corylus avellana) cotyledons. Physiol Plant 81:227–233CrossRefGoogle Scholar
  18. Gonzalez A, Arigita L, Majada J, Tames RS (1997) Ethylene involvement in in vitro organogenesis and plant growth of Populus tremula L. Plant Growth Regul 22:1–6CrossRefGoogle Scholar
  19. Higuchi M, Pischke MS, Mähönen AP, Miyawaki K, Hashimoto Y, Seki M, Kobayashi M, Shinozaki K, Kato T, Tabata S, Helariutta Y, Sussman MR, Kakimoto T (2004) In planta function of the Arabidopsis cytokinin receptor family. Proc Natl Acad Sci USA 101:8821–8826CrossRefPubMedGoogle Scholar
  20. Hoebee SE, Menn C, Rotach P, Finkeldey R, Holderegger R (2006) Spatial genetic structure of Sorbus torminalis: the extent of clonal reproduction in natural stands of a rare tree species with a scattered distribution. Forest Ecol Manage 226:1–8CrossRefGoogle Scholar
  21. Kepczynski J, Nemoykina A, Kepczynska E (2006) Ethylene and in vitro rooting of rose shoots. Plant Growth Regul 50:23–28CrossRefGoogle Scholar
  22. Letham DS, Palni LMS (1983) The biosynthesis and metabolism of cytokinins. Annu Rev Plant Physiol 34:163–197CrossRefGoogle Scholar
  23. Ludwig-Müller J, Vertocnik A, Town CD (2005) Analysis of indole-3-butyric acid-induced adventitious root formation on Arabidopsis stem segments. J Exp Bot 56:2095–2105CrossRefPubMedGoogle Scholar
  24. Ma JH, Yao JL, Cohen D, Morris B (1998) Ethylene inhibitors enhance in vitro root formation from apple shoot cultures. Plant Cell Rep 17:211–214CrossRefGoogle Scholar
  25. Malá J, Kálal J, Cvrčková H, Cvikrová M, Eder J (2000) The effect of reduction of exuded phenolic substances level on rooting of oak microcuttings. In: Cassels AC, Doyle BM, Curry PF (eds) Proceedings of the international symposium on methods and markers for quality assurance in micropropagation. Acta Hort 530:353–360Google Scholar
  26. Malá J, Máchová P, Cvrčková H, Čížková L (2005) Use of micropropagation for gene resources reproduction of noble deciduous species (Malus sylvestris, Pyrus pyraster, Sorbus torminalis, S. aucuparia and Prunus avium). Rep Forestry Res 50:219–224 (in Czech)Google Scholar
  27. Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  28. Nag S, Saha K, Choudhuri MA (2001) Role of auxin and polyamines in adventitious root formation in relation to changes in compounds involved in rooting. J Plant Growth Regul 20:182–194CrossRefGoogle Scholar
  29. Nordstrom AC, Eliasson L (1993) Interaction of ethylene with indole-3-acetic-acid in regulation of rooting in pea cuttings. Plant Growth Regul 12:83–90CrossRefGoogle Scholar
  30. Novák O, Hauserová E, Amakorová E, Doležal K, Strnad M (2008) Cytokinin profiling in plant tissues using ultra-performance liquid chromatography—tandem mass spectrometry. Phytochemistry 69:2214–2224CrossRefPubMedGoogle Scholar
  31. Oddou-Muratorio S, Le Guerroue B, Guesnet D, Demesure B (2001) Pollen- versus seed-mediated gene flow in a scattered forest tree species. Evolution 55:1123–1135PubMedGoogle Scholar
  32. Prat D, Daniel C (1993) Variabilité génétique l’Alisier torminal et du genere Sorbus. Rev For Fr 45:216–228Google Scholar
  33. Sabatini S, Beis Wolkenfelt H, Murfett J, Guilfoyle T, Malamy J, Benfey P, Leyser O, Bechtold N, Weisbeek P, Scheres B (1999) An auxin-dependent organizer of pattern and polarity in the Arabidopsis root. Cell 99:463–472CrossRefPubMedGoogle Scholar
  34. Scholten HJ (1998) Effect of polyamines on the growth and development of some horticultural crops in micropropagation. Sci Hort 77:83–88CrossRefGoogle Scholar
  35. Strnad M (1997) The aromatic cytokinins. Physiol Plant 101:674–688CrossRefGoogle Scholar
  36. Tarkowská D, Doležal K, Tarkowski P, Ǻstot C, Holub J, Fuksová K, Schmülling T, Sandberg G, Strnad M (2003) Identification of new aromatic cytokinins in Arabidopsis thaliana and Populus x canadensis leaves by LC-(+)ESI-MS and capillary liquid chromatography/frit-fast atom bombardment mass spectrometry. Physiol Plant 117:579–590CrossRefPubMedGoogle Scholar
  37. Werbrouck SPO, van der Jeugt B, Dewitte W, Prinsen E, Van Onckelen HA (1995) The metabolism of benzyladenine in S. floribundum Schott ‘Petite’ in relation to acclimatization problems. Plant Cell Rep 14:662–665CrossRefGoogle Scholar
  38. Werbrouck SPO, Strnad M, Van Onckelen HA, Debergh PC (1996) Meta-topolin, an alternative to benzyladenine in tissue culture? Physiol Plant 98:291–298CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Jana Malá
    • 1
  • Pavlína Máchová
    • 1
  • Helena Cvrčková
    • 1
  • Michal Karady
    • 2
  • Ondřej Novák
    • 2
  • Jaromír Mikulík
    • 2
  • Eva Hauserová
    • 2
  • Jarmila Greplová
    • 2
  • Miroslav Strnad
    • 2
  • Karel Doležal
    • 2
    Email author
  1. 1.Institute of Forest and Game ManagementJílovištěCzech Republic
  2. 2.Laboratory of Growth RegulatorsPalacký University, and Institute of Experimental Botany, AS CROlomouc-HoliceCzech Republic

Personalised recommendations