Experiments on protoplast fusion between triticum and secale
1991
Montezuma-de-Carbalho, J.M. | Tome, M.C. (University of Coimbra, Coimbra (Portugal). Instituto Botanico, Center for Plant Physiology and Cytology)
A first method suitable for the isolation of wheat protoplasts is described. Isolation of rye leaf protoplasts was also achieved. Embryos were detached and primordial leaves cut from surface-sterilized seeds of Triticum durum Desf. cultivar Celta and Secale cereale L., respectively. Digestion of the tissues was carried out with an enzyme mixture composed of cellulase RS (1 percent) and macerozyme R-10 (0.2 percent) for 14-16 h. These protoplasts were about 90 percent viable (as determined by fluorescein diacetate staining). It was not possible to distinguish between wheat and rye protoplasts by using nuclear staining with 5 percent modified carbol fuchsin. Protoplast fusion with polyethylene glycol was accomplished, but at a very low rate
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