No abstract

SummaryThe angiosperm family Asteraceae is characterized by composite inflorescences, which are highly organized structures consisting of different types of flowers. In order to approach the control of floral organ differentiation in Asteraceae at molecular level, we are studying regulation of flavonoid biosynthesis in Gerbera hybrida. Dihydroflavonol-4-reductase (dfr) expression is regulated according to anthocyanin pigmentation patterns in all tested gerbera varieties at several anatomical levels. We have isolated a promoter for one of the dfr genes, Pgdfr2. Gerbera plants transgenic for a Pgdfr2-uidA construct reveal that the activity of the Pgdfr2 promoter from one variety follows the pigmentation in other varieties which have different color patterns. It is thus evident that the observed complex regulation of dfr expression occurs in trans. In order to identify the trans-acting regulators, we isolated a cDNA (gmyc1) homologous to the previously characterized genes encoding bHLH-type regulators of the anthocyanin pathway in plants. The expression of gmyc1 in different varieties suggests that it has a major role in regulating dfr activity in corolla and carpel, but not in pappus and stamen. Specifically in gerbera, the identical patterns of gmyc1 and dfr expression in corolla tissue suggest that GMYC1 also regulates dfr expression in a region and flower type specific manner. Our studies show that in gerbera GMYC1-dfr interaction is part of several developmental processes characteristic for Asteraceae (such as specification of flower types across the composite inflorescence), whereas in other processes (such as differentiation of sepal as pappus) other regulators control dfr expression to determine the spatial specificity.

Abstract. Fluorescein isothiocyanate-conjugated dextran (FITC-dextran) is internalized by endocytosis into the lysosome-like vacuoles of Saccharomyces cerevisiae (Makarow, M., 1985, EMBO (Eur. Mol. Biol. Organ.) J. 4:1861-1866). Here we show that under energy depletion conditions FITC-dextran accumulated in a cytoplasmic compartment, from which it could be chased to the vacuole when the energy block was removed. The internal pH of the intermediate compartment under energy depletion was determined by fluorometry to be 5.8. The pH could be raised by the lysosomotropic agent ammonium chloride, the protonophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone (CCCP) and the ATPase inhibitors dicyclohexylcarbodiimide (DCCD) and sodium vanadate. The pH of the vacuole was found to be 6.5. It was raised by ammonium chloride, CCCP, and DCCD, but not with sodium vanadate. Efrapeptin had no effect on the internal pH of either compartment. By dissecting the endocytic pathway, two portions of the route leading to the vacuole could be studied separately. The internalization of FITC-dextran from the extracellular fluid to the intermediate compartment followed linear kinetics, was independent of energy, and occurred at temperatures of between 15 ° and 37°C. Transfer of the marker from the intermediate compartment to the vacuole required energy, took place at temperatures between 19 ° and 37°C, and had a half-time of 7 min at 37°C. Transport of the marker from the exterior of the cell to the vacuole did not require acidic pH values in the intermediate compartment or the vacuole. We suggest that the cytoplasmic compartment revealed by FITC-dextran, under energy depletion, represents the equivalent of the endosomes of mammalian cells.

No abstract