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.
Similar content being viewed by others
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
References
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 142:543–551
Arigita L, Tames RS, Gonzales A (2003) 1-Methylcyclopropene and ethylene as regulators of in vitro organogenesis in kiwi explants. Plant Growth Regul 40:59–64
Bollmark M, Kubát B, Eliasson L (1988) Variation in endogenous cytokinin content during adventitious root formation in pea cuttings. J Plant Physiol 132:262–265
Casson SA, Lindsey K (2003) Genes and signaling in root development. New Phytol 158:11–38
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 16:58–62
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 211–226
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 213–220
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 53–59
De Klerk GJ, Hanečáková J, Jasik J (2001) The role of cytokinins in rooting of stem slices cut from apple microcuttings. Plant Biosyst 135:79–84
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 57:63–71
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 14:875–884
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 15:3737–3747
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)
Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42
Fišerová H, Kula E, Klemš M, Reinöhl V (2001) Phytohormones as indicators of the degree of damage in birch (Betula pendula). Biologia 56:405–409
Fusseder A, Ziegler P (1988) Metabolism and compartmentation of dihydrozeatin exogenously supplied to photoautotrophic suspension-cultures of Chenopodium rubrum. Planta 173:104–109
Gonzalez A, Rodriguez R, Tames RS (1991) Ethylene and in vitro rooting of hazelnut (Corylus avellana) cotyledons. Physiol Plant 81:227–233
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 22:1–6
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 101:8821–8826
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 226:1–8
Kepczynski J, Nemoykina A, Kepczynska E (2006) Ethylene and in vitro rooting of rose shoots. Plant Growth Regul 50:23–28
Letham DS, Palni LMS (1983) The biosynthesis and metabolism of cytokinins. Annu Rev Plant Physiol 34:163–197
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 56:2095–2105
Ma JH, Yao JL, Cohen D, Morris B (1998) Ethylene inhibitors enhance in vitro root formation from apple shoot cultures. Plant Cell Rep 17:211–214
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 530:353–360
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)
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol Plant 15:473–497
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 20:182–194
Nordstrom AC, Eliasson L (1993) Interaction of ethylene with indole-3-acetic-acid in regulation of rooting in pea cuttings. Plant Growth Regul 12:83–90
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 69:2214–2224
Oddou-Muratorio S, Le Guerroue B, Guesnet D, Demesure B (2001) Pollen- versus seed-mediated gene flow in a scattered forest tree species. Evolution 55:1123–1135
Prat D, Daniel C (1993) Variabilité génétique l’Alisier torminal et du genere Sorbus. Rev For Fr 45:216–228
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 99:463–472
Scholten HJ (1998) Effect of polyamines on the growth and development of some horticultural crops in micropropagation. Sci Hort 77:83–88
Strnad M (1997) The aromatic cytokinins. Physiol Plant 101:674–688
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 117:579–590
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 14:662–665
Werbrouck SPO, Strnad M, Van Onckelen HA, Debergh PC (1996) Meta-topolin, an alternative to benzyladenine in tissue culture? Physiol Plant 98:291–298
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.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Malá, J., Máchová, P., Cvrčková, H. et al. Micropropagation of Wild Service Tree (Sorbus torminalis [L.] Crantz): The Regulative Role of Different Aromatic Cytokinins During Organogenesis. J Plant Growth Regul 28, 341–348 (2009). https://doi.org/10.1007/s00344-009-9099-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00344-009-9099-2