Acid phosphatase mutants in Chlamydomonas: isolation and characterization by biochemical, electrophoretic and genetic analysis

Abstract

In order to isolate acid phosphatase mutants in the green alga Chlamydomonas reinhardi, a staining method for detecting the enzyme activity in colonies has been developed. The occurrence of more than one acid phosphatase brought about some difficulty in the selection of mutants. We have, however, found an original method of selection based on the differential heat sensitivity of the enzymes. After treatment of the wild-type strain with N-methyl-N'-nitro-N-nitrosoguanidine, two types of mutants were recovered, then analyzed by biochemical and electrophoretic methods. In the first class of mutants (P(1), P(2), P(3),...) a heat-stable acid phosphatase bound to cellular debris of the crude extract was missing. The mutant P(a), representing the second class of mutations, was lacking a soluble heat-sensitive enzyme. These mutations were genetically different and exhibited mendelian inheritance.