Fluorescence microscopy to study colonization of conidia and hyphae of Cochliobolus Sativus by soil microorganisms

Epi-fluorescence microscopy facilitated observations of the colonization of hyphae and conidia of C. sativus and other plant pathogenic fungi in soil. Of six fluorochromes examined, acridine orange and europium chelate effectively differentiated cells of bacteria and actinomycetes from the melanized fungal structures. Under u.v. radiation, the colonizing organisms fluoresced intensively while the pigmented conidia appeared dark. Periodic observations of hyphal fragments and conidia of C. sativus incubated in nonsterile soil revealed rapid colonization of their surface primarily by bacteria. Whereas up to 80% of the hyphal fragments of C. sativus were lysed after 32 days' incubation in soil, the conidia remained intact. However, disorganization of protoplasts and disappearance of conidial septa were evident after the fourth week. After 2, 3 and 5 weeks incubation of the plant pathogens in soil, no significant difference was noticed between microbial colonization of conidia on 8.0 μm pore size “Nucle-pore” membranes and those placed directly on the soil surface.