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Isolation and regeneration of protoplasts from leaf explants of Rhyncholaelia digbyana
African Journal of Biotechnology
A protocol for the isolation and regeneration of protoplasts from leaf explant of Rhyncholaelia digbyana is presented. The protoplasts were isolated using hemicellulase enzymes at 1.5, 2.25, and 3% (w/v), pectinase at 0.5 and 0.75% (w/v) and cellulase at 1 and 2% (w/v). Protoplast counting was carried out with a Neubauer camera and an optical microscope at 40X, and viability was determined with Evans blue dye at 0.025% (w/v). The protoplasts were cultivated following the standard plate method,doi:10.5897/ajb2018.16540 fatcat:jvoq2s2z3za55o6l2pilwnsfpa