Enzymatic Isolation of Protoplasts from Fern Protonemal Cells Stainable with a Fluorescent Brightener

概要

論文の詳細を見る
Protoplasts were successfully isolated for the first time from the filamentous protonemal cells of ferns after the cells were cultured in contact with both air and medium. Sterilized spores of Adiantum capillus-veneris and Pteris vittata were inoculated on a piece of nylon mesh (40- μm mesh) placed on a mat of polyester fibers which was soaked in liquid culture medium, and the spores were illuminated from above with continuous red light. Protonemal cells, exposed to the air during this procedure, could be stained with Calcofluor White, a dye that binds to cell walls. Protoplasts were easily isolated from these protonemal cells by digestion of the cell wall with cellulase and pectinase. A total of 0.8-1.9 × 10^4 and 0.6-2.0 × 10^4 protoplasts were obtained from protonemata that originated from 10 mg of dry spores of Adiantum and of Pteris, respectively. Viability, as judged by staining with fluorescein diacetate was more than 90% for both species. Staining with 4'6-diamidino-2-phenylindole (DAPI) revealed that about half of the protoplasts of both species contained a nucleus.
日本植物生理学会の論文