[Effect of metalaxyl on the synthesis of RNA, DNA and protein in Phytophthora nicotianae]. / Wirkung von Metalaxyl auf die Synthese von RNA, DNA und Protein in Phytophthora nicotianae.

Metalaxyl is used to control diseases caused by fungi of the order of the Perenosporales. We investigated the action of this fungicid eon nucleic acid and protein synthesis in liquid cultures of Phytophthora nicotianae. The uptake of 32P, 3H-uridine, 3H-thymidine and 14C-leucine as precursors of nuclei acid and protein synthesis by the mycelium was not inhibited by metalaxyl. RNA synthesis as indicated by 3H-uridine incorporation was strongly inhibited (about 80%) by 0.5 micrograms/ml of metalaxyl. The inhibition was visible already few minutes after addition of the toxicant. Since the inhibition of incorporation of 3H-thymidine into DNA and of 14C-leucine into protein became significant 2-3 hours later, we conclude that metalaxyl primarily interfers with RNA synthesis. Synthesis of ribosomal RNA is more affected (more than 90%) than that of tRNA (about 55%) and poly(A)-containing RNA. Since in the presence of actinomycin, in contrast to metalaxyl, protein synthesis is inhibited immediately as a consequence of complete inhibition of RNA synthesis and of the short life-time of mRNA, it is also evident that mRNA synthesis is less strongly inhibited, at least during the early period of metalaxyl action. The molecular mechanism of metalaxyl inhibition of the transcription process remains open. The fungicide did not inhibit the activity of a partially purified RNA polymerase isolated from the fungus. On the other hand, the RNA synthesis (14C-UTP-incorporation) by a cell homogenate and by isolated nuclear fractions was inhibited significantly. Possibilities of the molecular action of metalaxyl are discussed. The RNA synthesis of some plant systems (cell cultures of Lycopersicon peruvianum, isolated nuclei from the same cell cultures, purified RNA polymerase from Spinacia oleracea chloroplasts) was not inhibited by metalaxyl, not even at high concentrations.

Gene expression in cytokinin-and light-mediated plastogenesis of Cucurbita cotyledons: ribulose-1,5-bisphosphate carboxylase/oxygenase.

The effects of a cytokinin (N(6)-benzyladenine, BA) and light on plastogenesis have been studied in detached Cucurbita cotyledons using the key enzyme of photosynthetic CO2 fixation, ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCase), as an example of a coordinated program of plastid and nucleo-cytoplasmic gene expression. Treatment of etiolated cotyledons with either BA in darkness or in light or light alone results in a marked and correlated stimulation of enzyme activity, quantity and biosynthesis (in-vivo [(14)C]leucine incorporation into immunoprecipitated enzyme protein), indicating an increase of de-novo synthesis under the influence of the two effectors. Cell-free translation of non-polyadenylated (poly(A)(-))RNA in an Escherichia coli system and total RNA in a wheat-germ system likewise demonstrate a light and hormone-dependent increase in the amounts of translatable mRNAs for the large (LS) and small subunits (SS) of RuBPCase (among other polypeptides). Hybridisation of poly(A)(-)RNA with a nick-translated LS gene of spinach RuBPCase reveals also two-to three-fold BA-or light-induced enhancement of LS mRNA content. Indications for stimulation of SS mRNA transcription are derived from inhibitor experiments with cordycepin. We conclude that the observed stimulation of de-novo RuBPCase synthesis by cytokinin and by light can be referred to the level of mRNA transcription, but it remains open whether this is a primary action. Furthermore, our results indicate that the coaction of the two exogenous factors is additive at different steps of RuBPCase formation, indicating independent actions in the causal chain between effector-signal transduction and RuBPCase gene expression.

Synthesis of ribosomal RNA in ribosome-deficient plastids of the mutant "albostrians" of Hordeum vulgare L.

The nuclear gene-induced plastome mutant "albostrians" of Hordeum vulgare L. is characterized by a plastid ribosome-deficiency. This ribosome deficiency could be caused by the lack of or a defect in chloroplast RNA polymerase. However in our investigations we found an activity of chloroplast RNA polymerase in wild-type and mutant leaves of "albostrians" barley by (1) electron microscopic autoradio-graphy after in vivo labelling of RNA, (2) determination of RNA polymerase activity in isolated plastids, and (3) characterization of the newly synthesized RNA by electrophoresis in polyacrylamide gels. The genes of 23S and 16S rRNA are transcribed in mutant plastids and the RNAs are processed. From these results we conclude that the enzymes involved in transcription and processing of chloroplast rRNA are synthesized on cytoplasmic ribosomes and that the plastid ribosome deficiency in albostrians barley is probably not caused by a defective chloroplast DNA dependent RNA polymerase or by a mutation in the genes of 23S and 16S chloroplast rRNA.