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Cytokinins in Tobacco and Wheat Chloroplasts.
Occurrence and
Changes Due to Light/Dark Treatment1
Eva Benková,
Erwin Witters,
Walter Van Dongen,
Jan Kolá ,
Václav Motyka,
Betislav Brzobohatý,
Harry A. Van Onckelen, and
Ivana Machá ková*
Institute of Biophysics, Academy of Sciences of the Czech Republic,
Královopolská 135, 61265 Brno, Czech Republic (E.B., B.B.); Department of Biology, University of Antwerpen, Universiteitsplein 1, B2610 Wilrijk-Antwerpen, Belgium (E.W., W.V.D., H.A.V.O.); and Institute of Experimental Botany, Academy of Sciences of the Czech
Republic, Rozvojová 135, 16502 Praha 6, Czech Republic (J.K.,
V.M., I.M.)
Although cytokinins (CKs)
affect a number of processes connected with chloroplasts, it has never
been rigorously proven that chloroplasts contain CKs. We isolated
intact chloroplasts from tobacco (Nicotiana tabacum L. cv SR1) and wheat (Triticum aestivum L. cv Ritmo) leaves
and determined their CKs by liquid chromatography/tandem mass
spectroscopy. Chloroplasts from both species contained a whole spectrum
of CKs, including free bases (zeatin and isopentenyladenine), ribosides
(zeatin riboside, and isopentenyladenosine), ribotides (isopentenyladenosine-5 -monophosphate, zeatin
riboside-5-monophosphate, and dihydrozeatin
riboside-5-monophosphate), and N-glucosides (zeatin-N9-glucoside,
dihydrozeatin-N9-glucoside,
zeatin-N7-glucoside, and
isopentenyladenine-N-glucosides). In chloroplasts there
was a moderately higher relative amount of bases, ribosides, and
ribotides than in leaves, and a significantly increased level of
N9-glucosides of zeatin and dihydrozeatin.
Tobacco and wheat chloroplasts were prepared from leaves at the end of
either a dark or light period. After a dark period, chloroplasts
accumulated more CKs than after a light period. The differences were
moderate for free bases and ribosides, but highly significant for
glucosides. Tobacco chloroplasts from dark-treated leaves contained
zeatin riboside-O-glucoside and dihydrozeatin
riboside-O-glucoside, as well as a relatively high CK
oxidase activity. These data show that chloroplasts contain a whole
spectrum of CKs and the enzymatic activity necessary for their
metabolism.
1
This research was supported by the Grant Agency
of the Czech Republic (grant no. 206/96/K188 to I.M.) and by the
Kontakt by the Ministry of Education of the Czech Republic (grant no.
ES040 to B.B.).
*
Corresponding author; e-mail machackova{at}ueb.cas.cz; fax
420-2-20390456.
Plant Physiol. (1999) 121: 245-252
Copyright Clearance Center: 0032-0889/99/121//08
© 1999 American Society of Plant Physiologists
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