Balanced growth and morphogenesis of Histoplasma capsulatum in a defined synthetic medium

Abstract

We have characterized balanced growth of yeast and hyphal cells of Histoplasma capsulatum and cells during transition from yeast to hyphae in a newly developed synthetic medium, R3B3. Homogeneous populations of yeast at 37 degrees C and hyphae at 25 degrees C grew in this medium with generation times of 10h and 19h, respectively. The growth rates were exponential, as demonstrated by the kinetics of net increase in dry weight. Identical rates of net increase were observed for ribonucleic acid (RNA) and protein, indicating conditions which approached steady state growth. In addition, this defined medium facilitated incorporation of radioactive precursors into RNA and protein and thus will allow for future detailed studies of macromolecular synthesis. When the incubation temperature of growing yeast cells was switched from 37 degrees C to 25 degrees C, the growth rate decreased as indicated in the generation time (19 h). Although the kinetic rates of RNA and protein synthesis also decreased, these rates were slightly faster relative to that observed for the increase in dry weight. During the later stage(s) of the yeast to hyphae transition a marked increase and subsequent decrease was observed for both RNA and protein. After this period of time, the synthesis of RNA and protein proceeded at the steady state rates usually observed in hyphal cultures.