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Abstract: Mono- or homokaryotic isolates were isolated from different strains of Agaricus bisporus by using isozymes as genetic markers. The mycelium cultures of the incompatible combination in the mating crosses were used to prepare protoplasts by using 1 5% lywallzyme and the maximum number of protoplasts was 4 8 x 107/ml The regeneration was carried out on the compost extract media (CEM) with a 3-5% regeneration frequency. The protoplasts of the different strains were fused in the fusogen of 30% PEG 4000 containing CaCL2 and 0.6M sucrose at pH 85 and were regenerated on the CEM. The fusants expressing a normal mycelium growth rate were selected for isozyme electrophoresis to find the heterokaryotic fusants and the fusion frequency was 105 to 106 Some fusants re-established the fertility and expressed some characteristics of the parent strains.
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