N. Kangatharalingam et Jc. Priscu, ISOLATION AND VERIFICATION OF ANATOXIN-A PRODUCING CLONES OF ANABAENA-FLOS-AQUAE (LYNGB) DE BREB FROM A EUTROPHIC LAKE, FEMS microbiology, ecology, 12(2), 1993, pp. 127-130
We present a technique to isolate and confirm anatoxin-a producing clo
nes (single trichome-isolates) of Anabaena flos-aquae (Lyngb.) de Breb
. from blooms of this cyanobacterium. A single trichome is isolated fr
om a field sample and grown in ASM medium. Single trichomes are then i
solated from this culture and grown in ASM medium to produce single cl
one cultures. Mouse bioassay, and thin-layer chromatography (TLC) usin
g purified anatoxin-a as reference is then used to confirm the anatoxi
n-a producing clones. Using this methodology, Anabaena flos-aquae samp
les collected during July 1991 from Hebgen Lake, Montana, were found t
o contain only 8.7% anatoxin-a producing clones. This minor proportion
of anatoxin-a producing clones apparently accounts for the anatoxin-a
produced by the entire population of A. flos-aquae. Our technique is
simple and reproducible. A selected clone of A. flos-aquae that produc
es anatoxin-a and one that does not produce anatoxin-a were deposited
in the UTEX culture collection, University of Texas at Austin.