Composition of culture medium influences zoosporogenesis and differentiation of Aphanomyces cochlioides

Abstract

A modified medium was used to culture mycelium and produce a large quantity of zoospores of Aphanomyces cochlioides, a principal pathogen of damping-off disease of sugar beet and root rot disease of spinach. The semisolid medium consisted of 17 g corn meal agar (CMA) added with 4 g of yeast extract (YE) per liter of 50 mM phosphate buffer (pH 6.8–7.0). This medium supported the production of ca. 106 zoospores ml−1 in 6-day-old cultures, approximately 11-fold higher than the commonly used CMA (17 g CMA per liter of water, pH 6.0 ± 0.2). Although morphological characters of the zoospores produced from the hyphae grown on CMA and CMA + YE were almost similar, they contrasted their developmental strategy after encystment induced by mechanical agitation. Cystospores originating from the zoospores on CMA regenerated zoospores (>80%), while those from CMA + YE germinated (ca. 80%) and produced hyphae. Furthermore, 4–10% of the germinated cystospores on CMA + YE had double germ tubes. The soluble protein profiles of zoospores produced on CMA and on CMA + YE demonstrated that several proteins were either different or expressed differently. Our results suggest that the culture medium directly influences zoosporogenesis in A. cochlioides hyphae and the developmental strategy of the produced zoospores.