Theory and application of growth delay analysis of colony formation for evaluation of injured population of the stressed fungal conidia.

ABSTRACT

A new concept of injured population assessment is proposed, in which the size of the injured population in stressed mold spores is evaluated by analyzing the colony formation process on a solid agar medium. In this method, a small paper disc containing mold spores is placed on a subculture agar plate, and the linear increase in the radius of the colony formed by development from the spore is measured over time. Then, the principle of the previously reported growth delay analysis (GDA) method originally using a liquid medium is applied to obtain the integrated viable ratio (IV) of the stressed population from the delay time relative to the growth of the unstressed population. On the other hand, the viable ratio (V) to the initial value as the colony count obtained with the stressed culture is obtained; the difference between the logarithms of V and IV is determined as the log number of the injured population. Applying this analysis method to heated spores of Cladosporium sphaerospermum, we determined the size of the injured population that occurred. This method was considered to be effective as a new method for quantifying injured populations using a solid medium.