Analysis of the effect of effective factors in protoplast preparation of Fusarium oxysporum for using in protoplast fusion

Protoplast fusion as a transgenic system is an effective technique improving microorganism performances by making genetic changes. Today, exploitation of this technique has beenbeneficial in biocontrol of weeds. For filamentous fungi such as Fusarium species, protoplast preparation is the first step in the common genetic transformation process. For this objective, the Fusarium oxysporum isolates were isolated from Bromrape (Orobanche sp.) and were identified by morphological and molecular characteristics. Thereafter, the effects of various factors including medium and incubation time in spore preparation were evaluated. Furthermore, the effect of some factors on protoplast preparation and longevity including enzyme concentrations, type and concentration of osmotic stabilizers and lytic incubation times were also assessed. The results of this study demonstrated that Spezieller Nährstoffarmer Agarwas the best medium to prepare spores and the best incubation time for spore germination was 16 hours. In addition, concentrations of 15mM for Glucanex and20mM for Driselase and 5 hours incubation time were the best lytic enzyme complex and incubation time period, respectively. The KCl at the concentration of 0.6M was the best osmotic stabilizer in protoplast preparation and longevity.