A Shake Culture Method for Pythiaceae Applicable to Rapid, Small-Scale Assay of Vegetative Physiology

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Abstract
A method is described for growing uniform mycelial suspensions of Pythium ultimum and Phytophthora megasperma var. sojae in a defined medium in shake culture. Dry wt doubling times of these fungi were 5-6 h and about 12 h, respectively. Incorporation of radioactive leucine, adenine, glycerol and acetate into proteins, nucleic acids and lipids by 3 ml cultures were measured easily with 10 min pulse periods. These synthetic activities were blocked in a predictable fashion if the cell samples were pretreated for 10 min with a variety of inhibitors known to impair specific cytoplasmic and mitochondrial functions. The cultures were maintained easily, and after about 12 h from the seeding of fresh cultures they were ready for use. The rapid growth and the ease and reproducibility of sampling make possible short-term manipulations of small amounts of cell material as well as the rapid production of large amounts of uniform, actively growing mycelium.

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