Combination of driselase and lysing enzyme in one molar potassium chloride is effective for the production of protoplasts from germinated conidia of Fusarium verticillioides.
Various cell wall degrading enzymes and the protoplasting media were evaluated for the production of protoplast in Fusarium verticillioides. Among the various enzymes tested, driselase at 12.5 mg/ml in 1 M KCl protoplasting medium produced the maximum number of protoplast. Next to driselase, lysing enzyme at 10 mg/ml in 1.2 M MgSO4 protoplasting medium was found to be the second best enzyme for the production of protoplast. More interestingly, the combined use of driselase @ 12.5 mg/ml and lysing enzyme @ 10 mg/ml in 1 M KCl exhibited the additive effect on protoplast formation. Germinated conidia of F. verticillioides are the most susceptible fungal material for protoplast production. The use of sucrose at 1.2 M in the regeneration medium supported the maximum regeneration of protoplast. From the present study, we recommend driselase (12.5 mg/ml) and lysing enzyme (10 mg/ml) in 1 M KCl protoplasting medium and germinated conidia of F. verticillioides for the maximum production of protoplasts and 1.2 M sucrose is the best osmoticum for the regeneration of protoplasts.
Department of Plant Pathology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Killikulam, Vallanadu, Tamil Nadu 628 252, India. Electronic address: rvrmoorthy@yahoo.com.
Vellaisamy
V
Ramamoorthy
Department of Plant Pathology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Killikulam, Vallanadu, Tamil Nadu 628 252, India.
Lavanya
L
Govindaraj
Department of Plant Pathology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Killikulam, Vallanadu, Tamil Nadu 628 252, India.
Madhumitha
M
Dhanasekaran
Department of Plant Pathology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Killikulam, Vallanadu, Tamil Nadu 628 252, India.
Sharmilee
S
Vetrivel
Department of Plant Pathology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Killikulam, Vallanadu, Tamil Nadu 628 252, India.
Krish K
KK
Kumar
Department of Plant Pathology, Agricultural College and Research Institute, Tamil Nadu Agricultural University, Killikulam, Vallanadu, Tamil Nadu 628 252, India.
Edward
E
Ebenezar
D002473:Cell Wall / Q000201:enzymology; D003470:Culture Media; D005656:Fungal Proteins / Q000378:metabolism*; D005670:Fusarium / Q000648:ultrastructure*; D006026:Glycoside Hydrolases / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology*; D011523:Protoplasts / Q000648:ultrastructure; D013172:Spores, Fungal / Q000502:physiology* / Q000648:ultrastructure*
D003470:Culture Media; D005656:Fungal Proteins; D011189:Potassium Chloride; D006026:Glycoside Hydrolases; C035212:driselase
Cell wall lytic enzyme; Osmotic stabilizers; Protoplast