Abstract
Antisera have been raised in rabbits against dihydrozeatin riboside and isopentenyladenosine, and their cross-reactivity characteristics have been examined in detail. These antisera, together with an antiserum previously raised against zeatin riboside, have been employed in radioimmunoassays. Separative procedures that enable a wide range of naturally occurring cytokinins to be separated prior to analysis by radioimmunoassay have been developed. The accuracy with which the following cytokinins can be quantified by our methods, which employ tritiated cytokinin recovery markers, has been estimated: zeatin riboside, zeatin, dihydrozeatin riboside, dihydrozeatin, O-glucosyl zeatin riboside, O-glucosyl zeatin, O-glucosyl dihydrozeatin riboside, O-glucosyl dihydrozeatin, zeatin-9-glucoside, zeatin-7-glucoside, lupinic acid, isopentenyladenosine, and isopentenyladenine.
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Badenoch-Jones J, Rolfe BG, Letham DS (1984a) Phytohormones,Rhizobium mutants and nodulation in legumes. V. Cytokinin metabolism in effective and ineffective pea root nodules. Plant Physiol 74:239–246
Badenoch-Jones J, Letham DS, Parker CW, Rolfe BG (1984b) Quantitation of cytokinins in biological samples using antibodies against zeatin riboside. Plant Physiol 75:1117–1125
Barthe GA, Stewart I (1985) Enzyme immunoassay (EIA) of endogenous cytokinins inCitrus. J Agric Food Chem 33:293–297
Bopp M, Gerhauser D, KeBler U (1986) On the hormonal system of mosses. In: Bopp M (ed) Plant growth substances 1985. Springer-Verlag, Berlin, pp 263–269
Brandon D, Hua S-ST, Corse JW, Yang SF, Layton LL (1980) Two rapid assays for cytokinins. Plant Physiol 65s:25
Cahill DM, Weste GM, Grant BR (1986) Changes in cytokinin concentrations in xylem extrudate following infection ofEucalyptus marginata Donn ex Sm withPhytophthora cinnamomi Rands. Plant Physiol 81:1103–1109
Chen CM, Ecker RL, McChesney JD (1976) N6-(Δ2-isopentenyl)adenosine from crown gall tumor tissue ofVinca rosea. Phytochemistry 15:1565
Constantinidou HA, Steel JA, Kozlowski TT, Upper CD (1978) Binding specificity and possible analytical applications of the cytokinin-binding antibody, anti-N6-benzyladenosine. Plant Physiol 62:968–974
De Greef W, Dekigel M, Hamers R (1980) A radioimmunoassay for cytokinins. Arch Int Physiol Biochim 88:B134–135
Eberle J, Arnscheidt A, Klix D, Weiler EW (1986) Monoclonal antibodies to plant growth regulators. III. Zeatin riboside and dihydrozeatin riboside. Plant Physiol 81:516–521
Erlanger BF, Beiser SM (1964) Antibodies specific for ribonucleosides and ribonucleotides and their reaction with DNA. Biochemistry 52:68–74
Ernst D (1986) Radioimmunoassay and gas chromatography/mass spectrometry for cytokinin determination. In: Linskens HF, Jackson JF (eds), Modern methods of plant analysis: New series, Vol. 3. Springer-Verlag, Berlin, pp 18–49
Ernst D, Schafer W, Oesterhelt D (1983a) Isolation and quantitation of isopentenyladenosine in an anise cell culture by single-ion monitoring, radioimmunoassay and bioassay. Planta 159:216–221
Ernst D, Schafer W, Oesterhelt D (1983b) Isolation and identification of a new naturally occurring cytokinin (6-benzylaminopurineriboside) from an anise cell culture (Pimpinella anisum L.). Planta 159:222–225
Fujii T, Ogawa N (1972) The absolute configuration of (−)dihydrozeatin. Tetrahedron Lett 3075–3078
Griggs PJ, Stuchbury T, Wang TL (1985) Analysis of cytokinin production inPhaseolus vulgaris using a radioimmunoassay for dihydrozeatin riboside. Abstracts of the 12th International Conference on Plant Growth Substances, August 26–31, Heidelberg, FRG, p 26
Hall PJ, Badenoch-Jones J, Parker CW, Letham DS, Barlow BA (1987) Identification and quantification of cytokinins in the xylem sap of mistletoes and their hosts in relation to leaf mimicry. Aust J Plant Physiolin press
Hansen CE, Wenzler H, Meins FJ (1984) Concentration gradients oftrans-zeatin riboside andtrans-zeatin in the maize stem. Plant Physiol 75:959–963
Hashizume T, Doi J, Sugiyama T (1986) Mass spectrometric determination of endogenous cytokinins ofAzolla. Agric Biol Chem 50:1035–1038
Hofman PJ, Forsyth C, Van Staden J (1985) A radioimmunoassay for dihydrozeatin and dihydrozeatin riboside, and its application to a study of thein vitro metabolism of dihydrozeatin by soybean callus. J Plant Physiol 121:1–12
Hofman PJ, Featonby-Smith BC, Van Staden J (1986a) The development of ELISA and IRA for cytokinin estimation and their application to a study of lunar periodicity inEcklonia maxima (Osbeck) Papenf. J Plant Physiol 122:455–466
Hofman PJ, Forsyth C, Van Staden J (1986b) The metabolism of kinetin and benzyladenine in soybean callus: Determination by radioimmunoassay and radioassay. J Plant Physiol 124:289–298
Horgan R, Hewett EW, Horgan JM, Purse J, Wareing PF (1975) A new cytokinin fromPopulus ×Robusta. Phytochemistry 14:1005–1008
Letham DS (1978) Cytokinins. In: Letham DS, Goodwin PB, Higgins TJ (eds), Phytohormones and related compounds: A comprehensive treatise, Vol. 1. Elsevier/North-Holland, Amsterdam, pp 205–263
Letham DS, Gollnow BI (1985) Regulators of cell division in plant tissues. XXX. Cytokinin metabolism in relation to growth and senescence of radish cotyledons. J Plant Growth Regul 4:129–145
Letham DS, Palni LMS (1983) The biosynthesis and metabolism of cytokinins. Annu Rev Plant Physiol 34:163–197
Letham DS, Wilson MM, Parker CW, Jenkins ID, MacLeod JK, Summons RE (1975) Regulators of cell division in plant tissues. XXIII. The identity of an unusual metabolite of 6-benzylaminopurine. Biochim Biophys Acta 399:61–70
MacDonald EMS, Akiyoshi DE, Morris RO (1981) Combined high performance liquid chromatography-radioimmunoassay for cytokinins. J Chromatogr 214:101–109
Milstone DS, Vold BS, Glitz DG, Shutt N (1978) Antibodies to N6-(Δ2-isopentenyl)adenosine and its nucleotide: Interaction with purified tRNAs and with bases, nucleosides and nucleotides of the isopentenyladenosine family. Nucleic Acids Res 5:3439–3455
Morris RO, Akiyoshi DE, MacDonald EMS, Morris JW, Regeir DA, Zaerr JB (1982) Cytokinin metabolism in relation to tumour induction byAgrobacterium tumefaciens. In: Wareing PF (ed), Plant growth substances. Academic Press, London, pp 175–183
Palmer MV, Wong OC (1985) Identification of cytokinins from xylem exudate ofPhaseolus vulgaris L. Plant Physiol 79:296–298
Palmer MV, Scott IM, Horgan R (1981) Cytokinin metabolism inPhaseolus vulgaris L. II. Comparative metabolism of exogenous cytokinins by detached leaves. Plant Sci Lett 22:187–195
Parker CW, Letham DS, Gollnow BI, Summons RE, Duke CC, MacLeod JK (1978) Regulators of cell division in plant tissues. XXV. Metabolism of zeatin by lupin seedlings. Planta 142:239–251
Randerath K, Randerath E (1973) Chemical characterization of unlabeled RNA and RNA derivatives by isotope derivative methods. In: Busch H (ed), Methods in cancer research, Vol 9. Academic Press, New York, p 3–69
Rosher PH, Jones HG, Hedden P (1985) Validation of a radioimmunoassay for (+)-abscisic acid in extracts of apple and sweet-pepper tissue using high-pressure liquid chromatography and combined gas chromatography-mass spectrometry. Planta 165:91–99
Sayavedra-Soto LA, Trione EJ, Durley RC (1983) Cytokinins in developing wheat spike by high performance liquid chromatography (HPLC)-radioimmunoassay (RIA). Plant Physiol 72s:114
Summons RE, Entsch B, Parker CW, Letham DS (1979a) Quantitation of the cytokinin glycoside complex of lupin pods by stable isotope dilution. FEBS Lett 107:21–25
Summons RE, Duke CC, Eichholzer JV, Entsch B (1979b) Mass spectrometric analysis of cytokinins in plant tissues. II. Quantitation of cytokinins inZea mays kernels using deuterium labelled standards. Biomed Mass Spectrom 6:407–413
Summons RE, Entsch B, Letham DS, Gollnow BI, MacLeod JK (1980) Regulators of cell division in plant tissues. XXVIII. Metabolites of zeatin in sweet-corn kernels: Purification and identifications using high-performance liquid chromatography and chemical-ionization mass spectrometry. Planta 147:422–434
Summons RE, Palni LMS, Letham DS (1983) Mass spectrometric analysis of cytokinins in plant tissues. IV. Determination of intact zeatin nucleotide by direct chemical ionization mass spectrometry. FEBS Lett 151:122–126
Tanaka Y, Abe H, Uchiyama M, Taya Y, Nishimura S (1978) Isopentenyladenine fromDictyostelium discoideum. Phytochemistry 17:543–544
Trione EJ, Krygier BB, Banowetz GM, Kathrein JM (1985) The development of monoclonal antibodies against the cytokinin zeatin riboside. J Plant Growth Regul 4:101–109
Tsui C, Shao LM, Wang CM, Tao G-Q, Letham DS, Parker CW, Summons RE, Hocart CH (1983) Identification of a cytokinin in water chestnuts (corms ofEleocharis tuberosa). Plant Sci Lett 32:225–231
Turnbull CGN, Hanke DE (1985) The control of bud dormancy in potato tubers. Measurement of the seasonal pattern of changing concentrations of zeatin-cytokinins. Planta 165:366–376
Van Onckelen H, Rudelsheim P, Hermans R, Horemans S, Mesens E, Hernalsteens J-P, Van Montagu M, De Greef J (1984) Kinetics of endogenous cytokinin, IAA and ABA levels in relation to the growth and morphology of tobacco crown gall tissue. Plant Cell Physiol 25:1017–1025
Vold BS, Leonard NJ (1981) Production and characterization of antibodies and establishment of a radioimmunoassay for ribosylzeatin. Plant Physiol 67:401–403
Vonk CR, Davelaar E, Ribot SA (1986) The role of cytokinins in relation to flower-bud blasting inIris cv. Ideal: Cytokinin determination by an improved enzyme-linked immunosorbent assay. J Plant Growth Regul 4:65–74
Walton D, Dashek W, Galston E (1979) A radioimmunoassay for abscisic acid. Planta 146:139–145
Weiler EW (1980) Radioimmunoassays fortrans-zeatin and related cytokinins. Planta 149:155–162
Weiler EW (1982) Plant hormone immunoassay. Physiol Plant 54:230–234
Weiler EW, Spanier K (1981) Phytohormones in the formation of crown gall tumors. Planta 153:326–337
Woodsworth ML, Latimer LJP, Janzer JJ, McLennan BD, Lee JS (1983) Characterization of monoclonal antibodies specific for isopentenyl adenosine derivatives occurring in transfer RNA. Biochem Biophys Res Commun 114:791–796
Zavala ME, Brandon DL (1983) Localization of a phytohormone using immunocytochemistry. J Cell Biol 97:1235–1239
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Badenoch-Jones, J., Parker, C.W. & Letham, D.S. Use of isopentenyladenosine and dihydrozeatin riboside antibodies for the quantification of cytokinins. J Plant Growth Regul 6, 159–182 (1987). https://doi.org/10.1007/BF02303051
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DOI: https://doi.org/10.1007/BF02303051