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Plant and Cell Physiology, 1986, Vol. 27, No. 7 1255-1261
© 1986

Article

Isolation of Hair Cells from Azolla filiculoides var. japonica Leaves

Eiji Uheda

Radiation Center of Osaka Prefecture Shinke-cho, Sakai, Osaka 593, Japan

A procedure has been developed to isolate hair cells from Anabaena containing packets of the aquatic fern Azolla. Unbroken algal packets, isolated by enzyme treatment and flotation on 12.5% Percoll solution, contained hair cells, Anabaena filaments and the envelope membrane. When the packet suspension was gently pipetted, hair cells attached to the envelope membrane became detached from the Anabaena filaments. Hair cells were separated by flotation on 34% Percoll solution and further purified by sifting through nylon mesh. Both branched and unbranched cells were isolated and about 70% remained unstained by 0.1% trypan blue. Electron microscopic observation showed that these cells were protoplasts enclosed by a thin envelope and had well preserved internal structures. The activities of ammonia-assimilating enzymes in the hair cells were much higher than those in Azolla leaves, while the activities in the endophytes were repressed to very low levels. These results suggest that hair cells play an important role in the assimilation of the nitrogen which the endophytes fix and release into the cavity.

(Received March 24, 1986; Accepted June 28, 1986)
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E. Uheda, K. Maejima, and N. Shiomi
Localization of Glutamine Synthetase Isoforms in Hair Cells of Azolla Leaves
Plant Cell Physiol., August 15, 2004; 45(8): 1087 - 1092.
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