Action of Kinetin on Cotyledons of Fenugreek

Isolated cotyledons of fenugreek (Trigonella foenum graecum L.), which respond rapidly and specifically to the application of cytokinins with stimulated expansion, have been used to study the primary action of kinetin. Gross chemical analysis showed that ribonucleic acid increased within 24 hours in response to kinetin application. 8-Azaguanine inhibited both kinetin-induced expansion and RNA synthesis; 5-fluorodeoxyuridine inhibited only the RNA synthesis.Cotyledons produced nitrate reductase activity in response to 20 mm nitrate only in the presence of either light or kinetin and especially in the presence of both. Abscisic acid and inhibitors of RNA and protein synthesis depressed this response. Inhibitors affecting chloroplast development and function did not reduce the response in the presence of light and kinetin.In vitro incorporation of (14)C-l-leucine and (14)C-l-phenyl-alanine into protein by various recombinations of microsomal and 160,000g supernatant fractions varied according to the pretreatment which the cotyledons had received before the preparation of the fractions. Stimulatory effects were mainly associated with the microsomal fractions.The formation of leucine-, valine-, and tyrosine-tRNA complexes by high speed supernatant fractions from differently pretreated cotyledons was also compared. The sharp stimulation of the process by adding tRNA was found to be independent of the kind of preincubation that the cotyledons used for the tRNA extraction had received.It is concluded that the evidence is not in favor of kinetin correcting specific tRNA deficiencies. Kinetin removes a limitation that prevents the synthesis of RNA and genome expression.