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cis-Isomers of Cytokinins Predominate in Chickpea Seeds throughout Their Development1

Robert Joseph Neil Emery*, Laurent Leport, Joanne Edith Barton, Neil Clifford Turner, and Craig Anthony Atkins

Centre for Legumes in Mediterranean Agriculture (R.J.N.E., L.L., C.A.A., J.E.B., N.C.T.), and Botany Department (R.J.N.E., C.A.A.), University of Western Australia, Nedlands, Western Australia, 6907, Australia; and University of Western Australia, Nedlands, Western Australia, 6907, AustraliaCommonwealth Scientific and Industrial Research Organization, Division of Plant Industry, Centre for Mediterranean Agricultural Research, Private Bag, P.O. Wembley, Western Australia, 6014, Australia (N.C.T.)

Trans-isomers of cytokinins (CK) are thought to predominate and have greater biological activity than corresponding cis-isomers in higher plants. However, this study demonstrates a system within which the predominant CK are cis-isomers. CK were measured at four developmental stages in developing chickpea (Cicer arietinum L. cultivar Kaniva) seeds by gas chromatography-mass spectrometry. Concentrations were highest at an early endospermic fluid stage and fell considerably when the cotyledons expanded. The cis-isomers of zeatin nucleotide ([9R-MP]Z), zeatin riboside ([9R]Z), and zeatin (Z) were present in greater concentrations than those of corresponding trans-isomers: (trans)[9R-MP]Z, (trans)[9R]Z, (trans)Z, or dihydrozeatin riboside. Dihydrozeatin, dihydrozeatin nucleotide, and the isopentenyl-type CK concentrations were either low or not detectable. Root xylem exudates also contained predominantly cis-isomers of [9R-MP]Z and [9R]Z. Identities of (cis)[9R]Z and (cis)Z were confirmed by comparison of ion ratios and retention indices, and a full spectrum was obtained for (cis)[9R]Z. Tissues were extracted under conditions that minimized the possibility of RNase hydrolysis of tRNA following tissue disruption, being a significant source of the cis-CK. Since no isomerization of (trans)[2H]CK internal standards occurred, it is unlikely that the cis-CK resulted from enzymic or nonenzymic isomerization during extraction. Although quantities of total CK varied, similar CK profiles were found among three different chickpea cultivars and between adequately watered and water-stressed plants. Developing chickpea seeds will be a useful system for investigating the activity of cis-CK or determining the origin and metabolism of free CK.

1   This research was funded by the Australian Cooperative Research Centre for Legumes in Mediterranean Agriculture and the Grains Research and Development Corporation of Australia.
*   Corresponding author; e-mail rjnemery{at}cyllene.uwa.edu.au; fax 61-8-9380-1001.

Plant Physiol. (1998) 117: 1515-1523
Copyright Clearance Center:   0032-0889/98/117//09
© 1998 American Society of Plant Physiologists




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