Abstract
The nonallelicgib-1 andgib-3 tomato (Lycopersion esculentum Mill.) mutants are gibberellin deficient and exhibit a dwarfed growth habit. Previous work has shown that this dwarfed growth pattern can be reversed by the application of a number of gibberellins and their precursors, includingent-kaurene (ent-kaur-16-ene). This indicates that they are blocked in gibberellin biosynthesis at a step prior toent-kaurene metabolism. The normal accumulation of carotenoids observed in these mutants suggests a functionally normal isoprenoid pathway.Ent-kaurene is synthesized from geranylgeranyl pyrophosphate in a two-step process with copalyl pyrophosphate as an intermediate.In vitro assays using young fruit extracts from wild-type andgib-2 plants resulted in the conversion of geranylgeranyl pyrophosphate to copalyl pyrophosphate, and the conversion of copalyl pyrophosphate toentkaurene. Similar assays usinggib-1 plants indicated a reduced ability for synthesis of copalyl pyrophosphate from geranylgeranyl pyrophosphate, and thus a reducedent-kaurene synthetase A activity. Furthermore,gib-3 extracts demonstrated a reduced ability to synthesizeent-kaurene from copalyl pyrophosphate, and thus a reducedent-kaurene synthetase B activity. These results establish the enzymatic conversion of geranylgeranyl pyrophosphate to copalyl pyrophosphate, and copalyl pyrophosphate toent-kaurene, as the sites of the mutations ingib-1 andgib-3 tomatoes, respectively. We also note that tomato fruit extracts contain components which are inhibitory toent-kaurene synthesis.
Similar content being viewed by others
References
Bohner J, Hedden P, Bora-Haber E, Bangerth F (1988) Identification and quantitation of gibberellins in fruits ofLycopersicon esculentum, and their relationship to fruit size inL. esculentum andL. pimpinellifolium. Physiol Plant 73:348–353
Coates RM, Ley DA, Cavendar PL (1978) Synthesis and carbon-13 nuclear magnetic resonance spectra ofalltrans-geranylgeraniol and its nor analogues. J Organ Chem 43:4915–4922
Coolbaugh RC (1983) Early steps of gibberellin biosynthesis. In: Crozier A (ed) The biochemistry and physiology of gibberellins, 1983. Praeger Publishers, New York, pp 53–98
Davisson VJ, Woodside AB, Poulter CD (1985) Synthesis of allylic and homoallylic isoprenoid pyrophosphates. Methods Enzymol 110:130–144
Duncan JD, West CA (1981) Properties of kaurene synthetase fromMarah macrocarpus endosperm: Evidence for the participation of separate but interacting enzymes. Plant Physiol 68:1128–1134
Fall R, West CA (1971) Purification and properties of kaurene synthetase fromFusarium moniliforme. J Biol Chem 246:6913–6928
Frost RG, West CA (1977) Properties of kaurene synthetase fromMarah macrocarpus. Plant Physiol 59:22–29
Gafni Y, Shechter I (1981) Isolation of a kaurene synthetase inhibitor from castor bean seedlings and suspension cultures. Plant Physiol 67:1169–1173
Graebe JE (1987) Gibberellin biosynthesis and control. Annu Rev Plant Physiol 38:419–465
Graebe JE, Dennis DT, Upper CD, West CA (1965) Biosynthesis of gibberellins: I. The biosynthesis of (−)-kaurene, (−)-kauren-19-ol, and trans-geranylgeraniol in endosperm nucellus ofEchinocystis macrocarpa Greene. J Biol Chem 240:1847–1854
Gray JC (1987) Control of isoprenoid biosynthesis in higher plants. Adv Bot Res 14:25–91
Groot SPC, Bruinsma J, Karssen CM (1987) The role of endogenous gibberellin in seed and fruit development of tomato: Studies with a gibberellin-deficient mutant. Physiol Plant 71:184–190
Hedden P, Phinney BO (1979) Comparison ofent-kaurene andent-isokaurene synthesis in cell-free systems from etiolated shoots of normal anddwarf-5 maize seedlings. Phytochemistry 18:1475–1479
Ingram TJ, Reid JB, Murfet IC, Gaskin P, Willis CL, MacMillian J (1984) Internode length inPisum. TheLe gene controls the 3β-hydroxylation of gibberellin A20 to gibberellin A1. Planta 160:455–463
Kobayashi M, Sakurai A, Saka H, Takahashi N (1989) Quantitative analysis of endogenous gibberellins in normal and dwarf cultivars of riee. Plant Cell Physiol 30:963–969
Murakami Y (1972) Dwarfing genes in rice and their relation to gibberellin biosynthesis. In: Carr DJ (ed) Plant growth substances, 1970. Springer-Verlag, Berlin, Heidelberg, New York, pp 166–174
Phinney BO (1984) Gibberellin A, dwarfism and the control of shoot elongation in higher plants. In: Crazier A, Hillman JR (eds) The biosynthesis and metabolism of plant hormones. Cambridge Society of Experimental Biology Seminar Series 23:17–41
Potts WC, Reid JB (1983) Internode length inPisum. III. The effect and interaction of theNa/na andLe/le gene differences on endogenous gibberellin-like substances. Physiol Plant 57:448–454
Shechter I, West CA (1969) Biosynthesis of gibberellins. V. Biosynthesis of cyclic diterpenoids from trans-geranylgeranyl pyrophosphate. J Biol Chem 244:3200–3209
Shen-Miller J, West CA (1982) Ent-kaurene biosynthesis in extracts ofHelianthus annuus L. seedlings. Plant Physiol 69:637–641
Turnbull CGN, Crozier A, Schwenen L, Graebe JE (1985) Conversion of [14C]gibberellin A12-aldehyde to C19- and C20-gibberellins in a cell-free system from immature seed ofPhaseolus coccineus L. Planta 165:108–113
Upper CD, West CA (1967) Biosynthesis of gibberellins II. Enzymatic cyclization of geranylgeranyl pyrophosphate to kaurene. J Biol Chem 242:3285–3292
Zeevaart JAD (1986) Characterization of three single gene dwarf mutants of tomato. J Cell Biochem S10B:33
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bensen, R.J., Zeevaart, J.A.D. Comparison ofEnt-kaurene synthetase A and B activities in cell-free extracts from young tomato fruits of wild-type andgib-1, gib-2, andgib-3 tomato plants. J Plant Growth Regul 9, 237–242 (1990). https://doi.org/10.1007/BF02041969
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF02041969