INFLUENCES OF EXPLANT TYPE AND ENZYME INCUBATION ON ISOLATED PROTOPLAST DENSITY AND VIABILITY IN TWO GARLIC CULTIVARS

Abstract

The present study reports on optimizing protoplast isolation and fusion in two garlic cultivars ‘Balady’ and ‘Seds 40’. Protoplast density and viability were investigated in four different explants (etiolated and green parts of the pseudostem and lower and upper parts of the leaves) under enzyme incubation for 1, 2, 3 and 4 h. Among different explants, used for protoplast isolation in Balady cultivar, the upper and lower parts of the leaves produced the highest number of total protoplasts (70 and 66 pps/0.1 ml) at 4 and 3 h enzyme incubation, respectively. However, the etiolated part of pseudostem produced the highest number of viable protoplast in which 52.5 pps/0.1 ml were obtained at 3 h enzyme incubation. For protoplast isolation in Seds 40 cultivar, the highest number of total protoplasts (125 and 107.5 pps/0.1 ml) as well as viable protoplasts (105 and 107.5 pps/0.1 ml) was obtained from the etiolated and the green parts of pseudostem, respectively. The cultivar ‘Seds 40’ yielded higher total and viable protoplasts than Balady cultivar. Isolated protoplasts of ‘Seds 40’ and ‘Balady’ were fused successfully at a protoplast density of 1 × 105 using either physical and/or electrical method. Optimization of the source of plant material as well as protoplast isolation conditions for garlic is a crucial step towards a successful protoplast fusion and subsequent colony formation.