Several forms of microbodies have been characterized on the basis of their biochemical functions. We have investigated cucumber cotyledons which house two different microbody forms during their development. In these cells, a shift from organelles with the enzymes of beta-oxidation and glyoxylate cycle to peroxisomes with the enzymes of the photosynthetic C2-cycle can be induced by light. The transition state and the time course of changes was studied at different levels of gene expression during the first 2 days of illumination, by quantifying the rate of de novo protein synthesis in cotyledons and by measuring the mRNA activities in vitro. Synthesis and turnover of particular proteins were determined during the transition stage by immunoprecipitation of malate synthase, isocitrate lyase, catalase, multifunctional protein, and thiolase, and quantified by fluorography. From the mRNA activities and the rate of protein synthesis, gene expression for enzymes of the glyoxylate cycle and beta-oxidation started to decrease 24-36 h after onset of continuous light. At that time the rate of synthesis of glycolate oxidase, a leaf peroxisomal marker, is already maximal. By pulse-chase experiments 0-48 h after the onset of light the speed and intensity of protein turnover were measured. Rates of proteolytic degradation of individual enzymes indicated that the different enzymes were not lost simultaneously or all at once. This excludes a destruction of the whole organelle by the lytic compartment.

Biochemie, Fachbereich Chemie, Universität Marburg, Hans-Meerwein-Straße, D-3550 Marburg, Bundesrepublik Deutschland Z. Naturforsch. 45c, Isocitrate Lyase c DNA, Malate Synthase c DNA, Glyoxysome, Leaf Peroxisome, Transition of Organelles Glyoxysomes are being replaced by leaf-type peroxisom es during the greening of dark-grown cucumber cotyledons. Light functions in this process as negative modulator of the gene expression of glyoxylate cycle enzymes but as positive regulator for the activation of glycollate oxidase formation. The differential gene expression was investigated at the level of m RNA amounts using c DNA probes hybridizing with malate synthase m RNA, isocitrate lyase m RNA, and glycollate oxidase m RNA. Hybrid ization probes were obtained from a c DNA library complementary to the germinationspecific m RNA s of cucumber cotyledons. The process of replacem ent of glyoxysomal proteins by leaf peroxisom al proteins was reversed to a certain extend when greened cotyledons were brought back in the dark. Hybridization on Northern b lots provided evidences that in greened cotyledons the amount of malate synthase m RNA and isocitratelyase m RNA starts to increase up on dark treatment.