Journal of Plant Growth Regulation

, Volume 31, Issue 3, pp 392–405 | Cite as

Involvement of cis-Zeatin, Dihydrozeatin, and Aromatic Cytokinins in Germination and Seedling Establishment of Maize, Oats, and Lucerne

  • Wendy A. Stirk
  • Kateřina Václavíková
  • Ondřej Novák
  • Silvia Gajdošová
  • Ondřej Kotland
  • Václav Motyka
  • Miroslav Strnad
  • Johannes van Staden
Article

Abstract

The aims of this study were to monitor endogenous cytokinin levels during germination and early seedling establishment in oats, maize, and lucerne to determine which cytokinin forms are involved in these processes; to quantify the transfer ribonucleic acid (tRNA)-bound cytokinins; and to measure cytokinin oxidase/dehydrogenase (CKX) activity. Cytokinins were identified using UPLC-MS/MS. The predominant free cytokinins present in the dry seeds were dihydrozeatin-type (DHZ) in lucerne and maize and cZ-type (cis-zeatin) in oats. Upon imbibition, there was a large increase in cZ-type cytokinins in lucerne although the cZ-type cytokinins remained at high levels in oats. In maize, the high concentrations of DHZ-type cytokinins decreased prior to radicle emergence. Four tRNA-bound cytokinins [cis-zeatin riboside (cZR)>N6-(2-isopentenyl)adenosine (iPR), dihydrozeatin riboside (DHZR), trans-zeatin riboside (tZR)] were detected in low concentrations in all three species investigated. CKX activity was measured using an in vitro radioisotope assay. The order of substrate preference was N6-(2-isopentenyl)adenine (iP)>trans-zeatin (tZ)>cZ in all three species, with activity fluctuating as germination proceeded. There was a negative correlation between CKX activity and iP concentrations and a positive correlation between CKX activity and O-glucoside levels. As O-glucosides are less resistant to CKX degradation, they may provide a readily available source of cytokinins that can be converted to physiologically active cytokinins required during germination. Aromatic cytokinins made a very small contribution to the total cytokinin pool and increased only slightly during seedling establishment, suggesting that they do not play a major role in germination.

Keywords

cis-Zeatin Cytokinin oxidase/dehydrogenase Dihydrozeatin Lucerne Maize Oats tRNA degradation 

References

  1. Armstrong DJ (1994) Cytokinin oxidase and the regulation of cytokinin degradation. In: Mok DWS, Mok MC (eds) Cytokinins: chemistry, activity and function. CRC Press, Boca Raton, pp 139–154Google Scholar
  2. Bassil NV, Mok DWS, Mok MC (1993) Partial purification of a cis-trans-isomerase of zeatin from immature seed of Phaseolus vulgaris L. Plant Physiol 102:867–872PubMedGoogle Scholar
  3. Bewley JD (1997) Seed germination and dormancy. Plant Cell 9:1055–1066PubMedCrossRefGoogle Scholar
  4. Bilyeu KD, Laskey JG, Morris RO (2003) Dynamics of expression and distribution of cytokinin oxidase/dehydrogenase in developing maize kernels. Plant Growth Regul 39:195–203CrossRefGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  6. Dewar J, Taylor JRN, Berjak P (1998) Changes in selected plant growth regulators during germination in sorghum. Seed Sci Res 8:1–8CrossRefGoogle Scholar
  7. Dietrich JT, Kamínek M, Blevins DG, Reinbott TM, Morris RO (1995) Changes in cytokinins and cytokinin oxidase activity in developing maize kernels and the effects of exogenous cytokinin on kernel development. Plant Physiol Biochem 33:327–336Google Scholar
  8. Dobrev P, Motyka V, Gaudinová A, Malbeck J, Trávníčková A, Kamínek M, Vaňková R (2002) Transient accumulation of cis- and trans-zeatin type cytokinins and its relation to cytokinin oxidase activity during the cell cycle of synchronized tobacco BY-2 cells. Plant Physiol Biochem 40:333–337CrossRefGoogle Scholar
  9. Emery N, Atkins C (2006) Cytokinins and seed development. In: Basra AS (ed) Handbook of seed science and technology. Food Products Press, New York, pp 63–93Google Scholar
  10. Emery RJN, Leport L, Barton JE, Turner NC, Atkins CA (1998) cis-Isomers of cytokinins predominate in chickpea seeds throughout their development. Plant Physiol 117:1515–1523PubMedCrossRefGoogle Scholar
  11. Emery RJN, Ma Q, Atkins CA (2000) The forms and sources of cytokinins in developing white lupine seeds and fruits. Plant Physiol 123:1593–1604PubMedCrossRefGoogle Scholar
  12. Faiss M, Zalubilová J, Strnad M, Schmülling T (1997) Conditional expression of the ipt gene indicates a function for cytokinins in paracrine signalling in whole tobacco plants. Plant J 12:401–415PubMedCrossRefGoogle Scholar
  13. Frébortová J, Fraaije MW, Galuszka P, Šebela M, Peč P, Hrbáč J, Novák O, Bilyeu KD, English JT, Frébort I (2004) Catalytic reactions of cytokinin dehydrogenase: preferences for quinones as electron acceptors. Biochem J 380:121–130PubMedCrossRefGoogle Scholar
  14. 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 62:2827–2840PubMedCrossRefGoogle Scholar
  15. Galuszka P, Frébort I, Šebela M, Peč P (2000) Degradation of cytokinins by cytokinin oxidases in plants. Plant Growth Regul 32:315–327CrossRefGoogle Scholar
  16. Galuszka P, Popelková H, Werner T, Frébortová J, Pospišilová H, Mik V, Köllmer I, Schmülling T, Frébot I (2007) Biochemical characterization of cytokinin oxidase/dehydrogenase from Arabidopsis thaliana expressed in Nicotiana tabacum L. J Plant Growth Regul 26:255–267CrossRefGoogle Scholar
  17. Gu R, Fu J, Guo S, Duan F, Wang Z, Mi G, Yuan L (2010) Comparative expression and phylogenetic analysis of maize cytokinin dehydrogenase/osidase (CKX) gene family. J Plant Growth Regul 29:428–440CrossRefGoogle Scholar
  18. Holub J, Hanuš J, Hanke DE, Strand M (1998) Biological activity of cytokinins derived from ortho- and meta-hydroxybenzyladenine. Plant Growth Regul 26:109–115CrossRefGoogle Scholar
  19. Kamínek M, Vaněk T, Motyka V (1987) Cytokinin activities of N6-benzyladenosine derivatives hydroxylated on the side-chain phenyl ring. J Plant Growth Regul 6:113–120CrossRefGoogle Scholar
  20. Kamínek M, Motyka V, Vaňková R (1997) Regulation of cytokinin content in plant cells. Physiol Plant 101:689–700CrossRefGoogle Scholar
  21. Kasahara H, Takei K, Ueda N, Hishiyama S, Yamaya T, Kamiya Y, Yamaguchi S, Sakakibara H (2004) Distinct isoprenoid origins of cis- and trans-zeatin biosyntheses in Arabidopsis. J Biol Chem 279:14049–14054PubMedCrossRefGoogle Scholar
  22. Leubner-Metzger G (2006) Hormonal interactions during seed dormancy release and germination. In: Basra AS (ed) Handbook of seed science and technology. Food Products Press, New York, pp 303–341Google Scholar
  23. Maaß H, Klämbt D (1981) On the biogenesis of cytokinins in roots of Phaseolus vulgaris. Planta 151:353–358CrossRefGoogle Scholar
  24. Martin RC, Mok MC, Habben JE, Mok DWS (2001) A maize cytokinin gene encoding an O-glucosyltransferase specific to cis-zeatin. Proc Natl Acad Sci USA 98:5922–5926PubMedCrossRefGoogle Scholar
  25. Mok DWS, Mok MC (2001) Cytokinin metabolism and action. Annu Rev Plant Physiol Plant Mol Biol 52:89–118PubMedCrossRefGoogle Scholar
  26. Motyka V, Vaňková R, Čapková V, Petrášek J, Kamínek M, Schmülling T (2003) Cytokinin-induced upregulation of cytokinin oxidase activity in tobacco includes changes in enzyme glycosylation and secretion. Physiol Plant 117:11–21CrossRefGoogle Scholar
  27. Novák O, Tarkowski P, Tarkowská D, Doležal K, Lenobel R, Strnad M (2003) Quantitative analysis of cytokinins in plants by liquid chromatography-single-quadrupole mass-spectrometry. Anal Chim Acta 480:207–218CrossRefGoogle Scholar
  28. Novák O, Hauserová E, Amakorová P, Doležal K, Strnad M (2008) Cytokinin profiling in plant tissues using ultra-performance liquid chromatography-electrospray tandem mass spectrometry. Phytochemistry 69:2214–2224PubMedCrossRefGoogle Scholar
  29. Pertry I, Václavíková K, Depuydt S, Galuszka P, Spíchal L, Temmerman W, Stes E, Schmülling T, Kakimoto T, van Montagu MCE, Strnad M (2009) Identification of Rhodococcus fascians cytokinins and their modus operandi to reshape the plant. Proc Natl Acad Sci USA 106:929–934PubMedCrossRefGoogle Scholar
  30. Quesnelle PE, Emery RJN (2007) cis-Cytokinins that predominate in Pisum sativum during early embryogenesis will accelerate embryo growth in vitro. Can J Bot 85:91–103CrossRefGoogle Scholar
  31. Sakakibara H (2006) Cytokinins: Activity, biosynthesis and translocation. Annu Rev Plant Biol 57:431–449PubMedCrossRefGoogle Scholar
  32. Singh S, Sawhney VK (1992) Endogenous hormones in seeds, germination behaviour and early seedling characteristics in a normal and ogura cytoplasmic male sterile line of rapeseed (Brassica napus L.). J Exp Bot 43:1497–1505CrossRefGoogle Scholar
  33. Spíchal L, Rakova NY, Riefler M, Mizuno T, Romanov GA, Strnad M, Schmülling T (2004) Two cytokinin receptors of Arabidopsis thaliana, CRE1/AHK4 and AHK3, differ in their ligand specificity in a bacterial assay. Plant Cell Physiol 45:1299–1305PubMedCrossRefGoogle Scholar
  34. Stirk WA, Gold JD, Novák O, Strnad M, van Staden J (2005) Changes in endogenous cytokinins during germination and seedling establishment of Tagetes minuta L. Plant Growth Regul 47:1–7CrossRefGoogle Scholar
  35. Stirk WA, Novák O, Václavíková K, Tarkowski P, Strnad M, van Staden J (2008) Spatial and temporal changes in endogenous cytokinins in developing pea roots. Planta 227:1279–1289PubMedCrossRefGoogle Scholar
  36. Strnad M (1997) The aromatic cytokinins. Physiol Plant 101:674–688CrossRefGoogle Scholar
  37. Takagi M, Yokota T, Murofushi N, Ota Y, Takahashi N (1985) Fluctuation of endogenous cytokinin contents in rice during its life-cycle–quantification of cytokinins by selected ion monitoring using deuterium-labelled internal standards. Agric Biol Chem 49:3271–3277CrossRefGoogle Scholar
  38. 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 x canadensis leaves by LC-(+)ESI-MS and capillary liquid chromatography/frit-fast atom bombardment mass spectrometry. Physiol Plant 117:579–590PubMedCrossRefGoogle Scholar
  39. Van Staden J, Drewes FE (1991) The biological activity of cytokinin derivatives in the soybean callus bioassay. Plant Growth Regul 10:109–115CrossRefGoogle Scholar
  40. Villalobos N, Martin L (1992) Involvement of cytokinins in the germination of chick-pea seeds. Plant Growth Regul 11:277–291CrossRefGoogle Scholar
  41. Von Schwartzenberg K, Núñez 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

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Wendy A. Stirk
    • 1
  • Kateřina Václavíková
    • 2
    • 3
  • Ondřej Novák
    • 2
  • Silvia Gajdošová
    • 4
  • Ondřej Kotland
    • 3
  • Václav Motyka
    • 4
  • Miroslav Strnad
    • 2
    • 5
  • Johannes van Staden
    • 1
  1. 1.Research Centre for Plant Growth and Development, School of Biological and Conservation SciencesUniversity of KwaZulu-Natal PietermaritzburgScottsvilleSouth Africa
  2. 2.Laboratory of Growth RegulatorsPalacký University & Institute of Experimental Botany AS CROlomoucCzech Republic
  3. 3.Department of Biochemistry, Faculty of SciencePalacký UniversityOlomoucCzech Republic
  4. 4.Laboratory of Hormonal Regulations in PlantsInstitute of Experimental Botany AS CRPrague 6Czech Republic
  5. 5.Centre of the Region Haná for Biotechnological and Agricultural Research, Faculty of SciencePalacký UniversityOlomoucCzech Republic

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