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AN IMPROVED METHOD FOR OBTAINING AXENIC CLONES OF PLANKTONIC BLUE‐GREEN ALGAE 1 , 2
Author(s) -
Carmichael Wayne W.,
Gorham Paul R.
Publication year - 1974
Publication title -
journal of phycology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.85
H-Index - 127
eISSN - 1529-8817
pISSN - 0022-3646
DOI - 10.1111/j.1529-8817.1974.tb02706.x
Subject(s) - axenic , biology , algae , agar , bacteria , agar plate , microbiology and biotechnology , botany , cyanobacteria , food science , genetics
SUMMARY Axenic clones from 5 isolates of Anabaena flosaquae , 1 isolate of Microcystis acruginosa , and 1 isolate of Aphanizomenon flos‐aquae were obtained by a combination of steps that provided a 1000‐fold reduction in the bacteria‐algae ratio and permitted bacteria‐free filaments or cells to be isolated and grown from agar pour plates. The first step consisted of the addition of phenol to a dark‐treated culture to selectively reduce the numbers of actively growing bacteria while leaving the resting algal cells viable. The next steps involved washing the treated algal suspension on a Millipore filter pad or membrane followed by plating in washed agar containing buffered mineral medium plus vitamins and soil extract. The final steps consisted of incubating the agar pour plates, coring bacteria‐free filaments or cells, culturing the agar cores in a buffered mineral medium, and rigorously testing the resulting cultures for bacteriological contamination. Between 50 and 90% of the cores grew, and of these about 50% were judged axenic. The method, with appropriate adaptations, should be suitable for obtaining axenic clones of other freshwater and marine algae.