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JXB Advance Access originally published online on June 9, 2006
Journal of Experimental Botany 2006 57(10):2237-2247; doi:10.1093/jxb/erj190
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© The Author [2006]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

Cytokinins in the perianth, carpels, and developing fruit of Helleborus niger L.

Petr Tarkowski1 *, Danuse Tarkowská2, Ondrej Novák2, Snjezana Mihaljevic3, Volker Magnus3, Miroslav Strnad2 and Branka Salopek-Sondi3,{dagger}

1Umeå Plant Science Centre, Department of Forest Genetics and Plant Physiology, Umeå, Sweden
2Laboratory of Growth Regulators, Palacky University and IEB ASCR, Olomouc, Czech Republic
3Rudjer Boskovic Institute, Zagreb, Croatia

{dagger}To whom correspondence should be addressed. E-mail: salopek{at}irb.hr

Reproductive development in the Christmas rose (Helleborus niger L.) differs from that in commonly investigated model plants in two important aspects: (i) the perianth develops a photosynthetic system, after fertilization, and persists until seed ripening; and (ii) the ripe seed contains an immature embryo which continues to mature off the mother plant. The possible roles of cytokinins in these processes are investigated here by analysing extracts of the perianth and the carpels/maturing fruit prepared during anthesis and four stages of post-floral development. trans-Zeatin, dihydrozeatin, N6-({Delta}2-isopentenyl)adenine, and their ribosides were identified by tandem mass spectrometry. Single ion monitoring in the presence of deuterated internal standards demonstrated the additional presence of the corresponding riboside-5'-monophosphates, O-glucosides, and 9-glucosides, and afforded quantitative data on the whole set of endogenous cytokinins. Fruit cytokinins were mostly localized in the seeds. Their overall concentrations increased dramatically during early seed development and remained high for 6–8 weeks, until shortly before seed ripening (the last time point covered in this work). Overall cytokinin levels in the perianth did not change markedly in the period covered, but the level of N6-({Delta}2-isopentenyl)adenine-type cytokinins appeared to increase slightly and transiently during the greening phase. The perianths of unpollinated or depistillated flowers, which survived, but did not pass through the complete greening process, contained significantly less cytokinins than observed in fruit-bearing flowers. This suggests that perianth greening requires defined cytokinin levels and supports the role of the developing fruit in their maintenance.

Key words: Christmas rose, cytokinin identification and quantification, fruit and seed development, Helleborus niger L., perianth greening


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