ELECTROFUSION OF PROTOPLASTS OF 2 SPECIES OF PORPHYRA (RHODOPHYTA)

Protoplasts of Porphyra linearis were fused with those of P. miniata using an electrofusion technique. Newly isolated protoplasts derived from these species were cleaned and resuspended in a solution containing 0.8 M mannitol and 1 mM CaCl2. Fusion was achieved by applying a pulsed DC current of 130 V (BTX Optimizer: field strength 2.71 kV cm(-1), 103.2 V peak) following the alignment of protoplasts with an AC current of 6 V (field strength 0.29 kV cm(-1), 11.2 V peak). The fused protoplasts from the two species were then transferred to D-ll medium and micro-cultured at 7-10 degrees C, 12 : ($) over bar 1 ($) over bar 2, L : ($) over bar D with a photon flux density of 16 mu mol m(-2) s(-1) After 4 weeks some of the putative hybrids developed plantlets of a few cells.