Abstract
Benzyladenine (BAP) uptake and metabolism were characterized during the key stages of shoot organogenesis in leaf explants of Petunia ‘MD1’. Using leaf explant transfer experiments, it was shown that exposure to 2.2 μM BAP for 6, 8 or 10 days induced shoot formation on 27, 80 and 100% of the explants respectively, with a concomitant increase in the number of shoots per explant. BAP uptake and metabolism were characterized in leaf explants after 1, 3, 6 or 10 days exposure to [3H]BAP or 10 days exposure plus an additional 2 days on basal medium (10+2). BAP and 9-β-D-ribofuranosyl-BAP ([9R]BAP) were detected at days 1 and 3 only. Therefore, the BAP free base was not detectable during the shoot induction period between days 6 and 10, as defined by leaf transfer experiments. The BAP ribotide pool was largest on day 1 and decreased to day 10+2. It is possible that the BAP ribotide pool provided either the active cytokinin itself or acted as a short-term storage form for the active cytokinin in petunia shoot organogenesis. Other metabolites detected in petunia leaf tissue included 7-β-D-glucopyranosyl-BAP ([7G]BAP), 9-β-D-glucopyranosyl-BAP ([9G]BAP) and an unidentified metabolite C.
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Abbreviations
- BAP:
-
benzyladenine
- [7G]BAP:
-
7-β-D-glucopyranosyl-BAP
- [9G]BAP:
-
9-β-D-glucopyranosyl-BAP
- [9R]BAP:
-
9-β-D-ribofuranosyl-BAP
- [9R-5′P]BAP:
-
5′-monophosphate of [9R]BAP
- [9R-5′PP]BAP:
-
5′-diphosphate of [9R]BAP
- [9R-5′PPP]BAP:
-
5′-triphosphate of [9R]BAP
- TEA:
-
Triethylamine
References
Ausubel F, Bahnsen K, Hanson M, Mitchell A and Smith H (1980) Cell and tissue culture of haploid and diploid Petunia ‘Mitchell’. Plant Mol Biol Newsletter 1: 26–32
Burch LR and Stuchbury T (1987) Activity and distribution of enzymes that interconvert purine bases, ribosides and ribotides in the tomato plant and possible implications for cytokinin metabolism. Physiol Plant 69: 283–288
Chen C (1981) Biosynthesis and enzymic regulation of the interconversion of cytokinin. In: JGuern and CPeaud-Lenoel, eds. Metabolism and Molecular Activities of Cytokinins, pp 34–43 New York: Springer-Verlag
Christianson ML and Warnick DA (1983) Competence and determination in the process of in vitro shoot organogenesis. Dev Biol 95: 288–293
Christianson ML and Warnick DA (1985) Temporal requirement for phytohormone balance in the control of organogenesis in vitro. Dev Biol 112: 494–497
Christianson ML (1987) Causal events in morphogenesis. In: CEGreen et al. eds. Plant Tissue and Cell Culture, pp 45–55. New York: Alan R. Liss
Fosket DE (1980) Hormonal control of morphogenesis in cultured tissues. In: FSkoog, ed. Plant Growth Substances 1979, pp 362–369. New York: Springer-Verlag
Horgan R (1987) Plant growth regulators and the control of growth and differentiation in plant tissue cultures. In: CEGreen et al. eds. Plant Tissue and Cell Culture, pp 135–149. New York: Alan R. Liss
Izhar S and Zelcar A (1984) Cell, tissue and organ culture in Petunia. In: KCSink, ed. Petunia, pp 111–122. New York: Springer-Verlag
Laloue M (1977) Cytokinins: 7-glucosylation is not a prerequisite of the expression of their biological activity. Planta 134: 273–275
Laloue M, Pethe-Terrine C and Guern J (1981) Uptake and metabolism of cytokinins in tobacco cells: Studies in relation to the expression of their biological activities. In: JGuern and CPeaud-Lenoel, eds. Metabolism and Molecular Activities of Cytokinins, pp 80–96. New York: Springer-Verlag
Laloue M and Pethe C (1982) Dynamics of cytokinin metabolism in tobacco cells. In: PFWareing, ed. Plant Growth Substances 1982, pp 185–195. New York: Academic Press
Letham DS, Tao GQ and Parker CW (1982) An overview of cytokinin metabolism. In: PFWareing, ed. Plant Growth Substances 1982, pp 143–153. New York: Academic Press
Letham DS and Palni LMS (1983) The biosynthesis and metabolism of cytokinins. Ann Rev Plant Physiol 34: 163–197
Letham DS and Gollnow BI (1985) Regulators of cell division in plant tissues. XXX. Cytokinin metabolism in relation to radish cotyledon expansion and senescence. J Plant Growth Regul 4: 129–145
Mitchell A, Hanson M, Skvirsky R and Ausubel F (1980) Anther culture of Petunia: Genotypes with a high frequency of callus, root and plantlet formation. Z Pflanzenphysiol 100: 131–146
Murashige T and Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497
Nitsch JP (1968) Studies on the mode of action of auxins, cytokinins and gibberelins at the subcellular level. In: FWightman and GSetterfield, eds., Biochemistry and Physiology of Plant Growth Substances, pp 563–580. Ottawa: The Runge Press
Rao PS, Handro W and Harada H (1973) Hormonal control of differentiation of shoots, roots and embryos in leaf and stem cultures of Petunia inflata and Petunia hybrida. Physiol Plant 28: 458–463
Skoog F and Miller C (1957) Chemical regulation of growth and organ formation in plant tissues cultured in vitro. Symp Soc Exp Biol 11: 118–131
Smigocki AC and Owen LD (1988) Cytokinin gene fused with a strong promoter enhances shoot organogenesis and zeatin levels in transformed plant cells. Proc Nat Acad Sci USA 85: 5131–5135
Van derKrieken WM, Croes AF, Bardense GW and Wullems GJ (1988) Uptake and metabolism of benzyl adenine in the early stage of flower bud development in vitro in tobacco. Physiol Plant 74: 113–118
Van derKrieken WM, Croes AF, Smulders MJ and Wullems GJ (1990) Cytokinins and flower bud formation in vitro in tobacco: Role of the metabolites. Plant Physiol 92: 565–569
Wilson MM, Gordon ME, Letham DS and Parker CW (1974) Regulators of cell division in plant tissues. XIX. The metabolism of 6-benzylaminopurine in radish cotyledons and seedlings. J Exp Bot 87: 725–32
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This research was supported in part by NSF Grant DCB-8917378 to J.D.C. and USDA-CRGO Grant 89-37261-4791 to T.J.C.
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Auer, C.A., Laloue, M., Cohen, J.D. et al. Uptake and metabolism of benzyladenine during shoot organogenesis in Petunia leaf explants. Plant Growth Regul 11, 105–114 (1992). https://doi.org/10.1007/BF00024063
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DOI: https://doi.org/10.1007/BF00024063