PRODUCTION OF NEW SPECIES OF EDIBLE MUSHROOMS BY PROTOPLASTS FUSION METHOD .3. PROTOPLASTS FUSION AND ANALYSIS OF THE FUSANT BETWEEN PLEUROTUS-SAJOR-CAJU AND MYCOLEPTODONOIDES-AITCHISONII

Inter heterogeneric cell fusion using mononuclear auxotrophic mutants were performed between Pleurotus sajor-caju (Fr.) Singer and Mycolepto donoides aitchisonii (Berk.) Maas G. The optimum conditions of fusion in the individual electric fusion method, in a pearl chain electric fu sion method, and in a polyethyleneglycol method were investigated, and fusion regeneration rates were calculated. The individual electric fu sion (giant protoplast fusion) method had its largest value of 19.2% u nder conditions where a parallel electrode chamber, having 2 mm width was used to set a pulse rate of four in an AC (alternating current) el ectric field of 1 MHz and 30 V, and a DC (direct current) electric fie ld of 50 V and 20 mu sec. A 0.05 M glycine-NaOH buffer solution (pH 6. 5) containing 0.3 M of sorbitol and 0.05 M of CaCl2 was used between e lectrodes. With the testing of five fusants, the main results were as Follows: 1) A clamp-connection was observed in five strains, and the b inucleation due to fusion was proven. 2) A zone line was observed betw een both parental strains in all five strains.3) Following isozyme ana lysis, the same band as that of both parent strains was confirmed in t he bands of esterase, malate dehydrogenase and alcohol dehydrogenase o f the fusion strains. 4) The fusion strains were rated by 81% of the e xaminers, to have the aroma of M. aitchisonii.