Protoplast Preparation Conditions of Rhizoctonia cerealis and Pathogenicity of Regenerated Strains

Rhizoctonia cerealis strain HD-6was used for studying the protoplast preparation and regeneration,and the pathogenicity of wild-type and regenerated strains was compared.Four lytic enzymes,lywallzyme,driselase,snailase and cellulose,were used for the protoplast preparation independently.The effects of enzymolysis temperature,enzymolysis time and solution pH on protoplast preparation were investigated according to the single factor tests.Among the four enzymes,the driselase had the best efficiency for protoplast preparation and the protoplast quantity per gram of mycelium reached 2.6×108.The best conditions for the protoplast preparation were that the mycelium of R.cerealis was digested at 24℃in driselase solution(pH6.0)for 3 h.The prepared protoplasts could produce mycelium,which had the similar pathogenicity to that of the wild-type strain.