title: Factors affecting protoplast formation by Rhizoctonia solani.
abstract: Novozym 234 was the most frequently used enzyme for production of Rhizoctonia solani protoplasts. Since manufacture of this enzyme was discontinued in the late 1990s, a new procedure was developed by testing lytic enzymes from Sigma and by examining factors affecting protoplast formation. The combination of 20 mg/mL Driselase and 10mg/mL lysing enzyme was effective in releasing protoplasts from R. solani. The optimal condition for enzyme treatment of mycelium was incubation at 37 degrees C for 15 min followed by 34 degrees C for 105 min. The amount of protoplasts produced was positively correlated with growth rate and negatively correlated with mycelial density. Under favorable conditions, R. solani mycelia released 1.68 x 10(6) protoplasts/mL that is comparable with that produced with Novozym 234. Among various media tested, the best solid medium for protoplast regeneration was 1% V-8 juice agar, while the best liquid medium was 10% potato dextrose broth.
authors.affiliation: Department of Plant Pathology, National Chung Hsing University, Taichung, Taiwan.
authors.forename: Tung-Hsen
authors.initials: TH
authors.lastname: Liu
authors.forename: Mei-Ju
authors.initials: MJ
authors.lastname: Lin
authors.forename: Wen-Hsiung
authors.initials: WH
authors.lastname: Ko
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D002480:Cellulase / Q000378:metabolism; D004798:Enzymes / Q000378:metabolism; D005656:Fungal Proteins / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D025282:Mycelium / Q000378:metabolism; D011523:Protoplasts / Q000378:metabolism*; D012232:Rhizoctonia / Q000166:cytology / Q000502:physiology
chemical_list: D004798:Enzymes; D005656:Fungal Proteins; C045154:novozym 234; D006026:Glycoside Hydrolases; C035212:driselase; D002480:Cellulase