A simple rapid photometric estimation of the growth response of immobilized cells to fusaric acid and gamma radiation

Abstract

A simple and rapid photometric method was developed in order to evaluate the growth responses of cells to various factors in vitro. Cells of Asparagus officinalis L. were mixed with autoclaved agar at 40°C and 90 μl drops were dispensed into several Petri dishes. After the drops solidified, they were covered with a liquid growth medium and incubated on a gyratory shaker. Growth was measured every 2 or 3 days by two methods. In the first, the agar drop was placed under a stereomicroscope with substage illumination and the light passing through the embedded cells was measured by a light-meter. Growth, expressed by the increase in cell density, was inversely proportional to the intensity of transmitted light. In the second method, the agar drop was melted in a microwave oven and the packed cell volume was measured. The correlation between the two methods showed that the photometric method can be used to assess growth response of immobilized cells, during the first two weeks of culture. This method was used to evaluate growth responses to the toxin fusaric acid and to gamma radiation. The photometric method requires a small amount of inoculum, standard microscopic equipment and can be used to determine the effect of various factors on the growth of intact plant cells in vitro without disruption.