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Journal of Experimental Botany, Vol. 53, No. 378, pp. 2193-2200, November 1, 2002
© 2002 Oxford University Press

A trans-zeatin riboside in root xylem sap negatively regulates adventitious root formation on cucumber hypocotyls

Received 14 March 2002; Accepted 3 July 2002

Takeshi Kuroha1, Hisashi Kato2, Tadao Asami2, Shigeo Yoshida2, Hiroshi Kamada1 and Shinobu Satoh3,1

1 Institute of Biological Sciences, University of Tsukuba, Ibaraki 305-8572, Japan
2 RIKEN (The Institute of Physical and Chemical Research), 2-1 Hirosawa, Wako, Saitama 351-0198, Japan

Abbreviations: BA, 6N-benzyladenine; cis-Z, cis-zeatin; cis-ZR, cis-zeatin riboside; MS, mass spectrometry; MS medium, Murashige and Skoog’s medium; MS/MS, tandem mass spectrometry; m/z, mass-to-charge ratio; HPLC, high performance liquid chromatography; iPMP, isopentenyladenosine-5'-monophosphate; LC/MS/MS, liquid chromatography/tandem mass spectrometry; Z, trans-zeatin; ZMP, trans-zeatin riboside-5'-monophosphate; ZOG, trans-zeatin-O-glucoside; ZOGR, trans-zeatin-O-glucoside riboside; ZR, trans-zeatin riboside.

Shoot cultures of cucumber were used to analyse the roles of root-derived substances in adventitious root formation on hypocotyl tissues. Xylem sap collected from the roots of squash had a strong inhibitory effect on the formation of hypocotyl adventitious roots. Double-solvent extraction followed by fractionation with both normal and reverse phase column chromatographies and analysis by liquid chromatography/tandem mass spectrometry identified trans-zeatin riboside (ZR) as the primary suppressor of adventitious root formation. ZR was the predominant cytokinin present in the xylem sap, occurring at a concentration of 2x10–8 M. Application of ZR at concentrations from 3.16x10–9 M effected inhibition of adventitious root formation. These results suggest that ZR transported from roots via xylem sap may act as an endogenous suppressor of hypocotyl adventitious root formation in planta.

Key words: Key words: Adventitious root suppression, cucumber, Cucumis sativus, Cucurbita, cytokinin, squash, xylem sap, zeatin riboside.


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