A New Technique for Photobiont Culturing and Manipulation

Abstract

Comparisons of whole-lichen physiology to the respective photobionts have often been unclear due to inherent differences in isolated photobiont culturing techniques. The use of 13-mm-diameter cellulose-acetate discs allows photobiont cultures access to nutrient agar medium, while improving ease of manipulation and distinct separation from the agar. Adequate culture growth for experimentation is reached in approximately three weeks, a time comparable to standard nutrient agar and liquid cultures. These discs are then available for use in a variety of manipulative techniques. Chlorophyll determination of an entire algal disc culture is obtainable because the discs readily dissolve in dimethylsulphoxide (DMSO), with no interference in the 400–700 nm range. Photosynthesis and respiration may be measured with standard gas exchange equipment. Photobiont discs allow for fumigation in the gas phase with no increase in external ⊂pH reported to occur during gaseous fumigations in liquid media. The disc system is also useful for fluorescence studies. Trebouxia erici cultures exhibited a CO⊂2 gas exchange on a gram dry weight basis similar to whole lichen systems. The ease with which photobionts can be cultured and manipulated using this system allows for expanded experimentation and comparisons.