Endogenous Phytohormones in Spontaneously Regenerated Centaurium erythraea Rafn. Plants Grown In Vitro
- 339 Downloads
Phytohormones are important regulators of numerous developmental and physiological processes in plants. Spontaneous morphogenesis of the common centaury (Centaurium erythraea Rafn.) is possible on nutrition medium without addition of any plant growth regulator depending solely on endogenous phytohormone levels. Thus, this plant species represents a very good model system for the investigation of numerous physiological processes under phytohormonal control in vitro. We analysed the total amount of endogenous cytokinins (CKs) including the contents of their individual groups in shoots and roots of C. erythraea plants grown in vitro. The total amount of endogenous CKs was 1.4 times higher in shoots than in roots. Inactive or weakly active N-glucosides found to predominate in both organs of centaury plants, whereas free bases and O-glucosides represented only a small portion of the total CK pool. Consequently, centaury roots showed higher IAA content as well as IAA/free CK base ratios compared to shoots. Centaury tissues also showed increased levels of “stress hormones”. In contrast to SA, considerably higher levels of ABA were found in centaury shoots than in roots. Our results could serve as a basis for understanding and elucidating spontaneous de novo shoot organogenesis and further plant regeneration of C. erythraea in vitro.
KeywordsCommon centaury Root explants Hormone metabolism Cytokinins Auxins Plant development Stress hormones
Abbreviations for CKs Adopted and Modified According to Kamínek and Others (2000)
cis-zeatin 9-riboside O-glucoside
Dihydrozeatin 9-riboside O-glucoside
N 6-(∆2- isopentenyl)adenine 7-glucoside
N 6-(∆2- isopentenyl)adenine 9-glucoside
N 6-(∆2- isopentenyl)adenine 9-riboside
N 6-(∆2- isopentenyl)adenine 9-riboside-5′-monophosphate
trans-zeatin 9-riboside O-glucoside
This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (Grant No. ON173015) and the Czech Science Foundation (P506/11/0774).
M. Trifunović-Momčilov and M. Petrić contributed to all in vitro experiments. M. Trifunović-Momčilov and V. Motyka contributed in data analyses and manuscript preparation. J. Holík, J. Malbeck and P.I. Dobrev contributed to all experimental work considering endogenous plant hormones analyses. S. Jevremović contributed to all statistical analyses. I.Č. Dragićević contributed to data analyses and obtained result’s interpretation. A. Subotić supervised the whole study and also contributed in preparing the final manuscript.
Compliance with Ethical Standards
Conflict of interest
The authors declare that they have no conflict of interest.
- Gajdošová S, Spíchal L, Kamínek M, Hoyerová K, Novák O, Dobrev PI, Galuszka P, Klíma P, Gaudinová A, Žižková E, Hanuš J, Dančák M, Trávníček B, Pešek B, Krupička M, Vaňková R, Strnad M, Motyka V (2011) Distribution, biological activities, metabolism, and the conceivable function of cis-zeatin-type cytokinins in plants. J Exp Bot 62:2827–2840CrossRefPubMedGoogle Scholar
- Izumi K, Nakagawa S, Kobayashi M, Oshio H, Sakurai A, Takashi N (1988) Levels of IAA, cytokinins, ABA and ethylene in rice plants as affected by a gibberellin biosynthesis inhibitor, uniconazole-P. Plant Cell Physiol 29:97–104Google Scholar
- Jensen SR, Schripsema J (2002) Chemotaxonomy and pharmacology of Gentianaceae. In: Struve L, Albert V (eds) Gentianaceae: systematics and natural history. Cambridge University Press, Cambridge, pp 573–631Google Scholar
- Jones B, Gunneras SA, Petersson SV, Tarkowski P, Graham N, May S, Dolezal K, Sandberg G, Ljung K (2010) Cytokinin regulation of auxin synthesis in Arabidopsis involves a homeostatic feedback loop regulated via auxin and cytokinin signal transduction. Plant Cell 22:2956–2969CrossRefPubMedPubMedCentralGoogle Scholar
- Letham DS (1994) Cytokinins as phytohormones-sites of biosynthesis, translocation and function of translocated cytokinin. In: Mok DWS, Mok MC (eds) Cytokinins: chemistry, activity and function. CRC Press, Boca Raton, pp 57–80Google Scholar
- Liu J, Mehdi S, Topping J, Tarkowski P, Lindsey K (2010) Modelling and experimental analysis of hormonal crosstalk in Arabidopsis. Mol Syst Biol 6, Article number 373:1–13Google Scholar
- Miyawaki K, Tarkowski P, Matsumoto-Kitano M, Kato T, Sato S, Tarkowska D, Tabata S, Sandberg G, Kakimoto T (2006) Roles of Arabidopsis ATP/ADP isopentenyltransferases and tRNA isopentenyltransferases in cytokinin biosynthesis. Proc Natl Acad Sci USA 103:16598–16603CrossRefPubMedPubMedCentralGoogle Scholar
- Nordström A, Tarkowski P, Tarkowska D, Norbaek R, Astot C, Dolezal K, Sandberg G (2004) Auxin regulation of catokinin biosynthesis in Arabidopsis thaliana: a factor of potential importance for auxin-cytokinin-regulated development. Proc Natl Acad Sci USA 101:8039–8044CrossRefPubMedPubMedCentralGoogle Scholar
- Skoog F, Miller CO (1957) Chemical regulation of growth and organ formation in plant tissue cultured in vitro. Sym Soc Exp Biol 11:118–131Google Scholar
- Subotić A, Budimir S, Grubišić D, Momčilović I (2003/2004) Direct regeneration of shoots from hairy root cultures of Centaurium erythraea inoculated with Agrobacterium rhizogenes. Biol Plant 47:617–619Google Scholar
- Subotić A, Jevremović S, Grubišić D, Janković T (2009a) Spontaneous plant regeneration and production of secondary metabolites from hairy root cultures of Centaurium erythraea Rafn. In: Jain SM, Saxena PK (eds) Protocols for in vitro cultures and secondary metabolite analysis of aromatic and medicinal plants, methods in molecular biology, vol 547. Springer, Berlin, pp 205–217Google Scholar
- Trifunović M, Cingel A, Simonović A, Jevremović S, Petrić M, Dragićević I, Motyka V, Dobrev PI, Zahajská L, Subotić A (2013) Overexpression of Arabidopsis cytokinin oxidase/dehydrogenase genes AtCKX1 and AtCKX2 in transgenic Centaurium erythraea Rafn. Plant Cell Tissue Org 115:139–150CrossRefGoogle Scholar
- Trifunović M, Motyka V, Cingel A, Subotić A, Jevremović S, Petrić M, Holík J, Malbeck J, Dobrev PI, Dragićević I (2015) Changes in cytokinin content and altered cytokinin homeostasis in AtCKX1 and AtCKX2-overexpressing centaury (Centaurium erythraea Rafn.) plants grown in vitro. Plant Cell Tiss Org 120:767–777CrossRefGoogle Scholar
- Von Schwartzenberg K, Nunez MF, Blaschke H, Dobrev PI, Novák O, Motyka V, Strnad M (2007) Cytokinins in the bryophyte Physcomitrella patens: analyses of activity, distribution and cytokinin oxidase/dehydrogenase overexpression reveal the role of extracellular cytokinins. Plant Physiol 145:786–800CrossRefGoogle Scholar
- Werner T, Motyka V, Laucou V, Stems R, Van Onckelen H, Schmülling T (2003b) Cytokinin deficient transgenic Arabidopsis plant show multiple developmental alterations indicating opposite function of cytokinins in the regulation of shoot and root meristem sctivity. Plant Cell 15:2532–2550CrossRefPubMedPubMedCentralGoogle Scholar