不同因素对金针菇原生质体制备和再生的影响

Effects of Different Factors on the Protoplast Preparation and Reproduction in Flammulina Velutipes

比较了不同浓度裂解酶及组合、渗透压稳定剂、酶解时间等因素对金针菇原生质体得率的影响以及不同渗透压稳定剂、培养基、接种方法等因素对金针菇原生质体再生的影响。试验结果表明:固体培养10d的金针菇菌丝,以0.5mol/L KCl作渗透压稳定剂,加入1%纤维素酶和1%溶菌酶在25℃下酶解1.5h,分离原生质体效果最佳,原生质体产量可达27.8×105个/ml以上;以0.5mol/L KCl作渗透压稳定剂,在25℃条件下,金针菇原生质体采用直接涂布法接种在RCM培养基上培养,再生率最高为0.5%。

The effects of factors, such as lyric enzyme, enzyme concentration, osmotic stabilizers, duration of hyphae digested with lytic enzyme have been studied on the formation of protoplasts in F. Velutipes. Maximum yield of protoplasts could be obtained with 10d - mycelia by using 1% cellulase and 1% lysozyme stabilized with 0.5mol/L KCl and digesting for 1.5h at 25℃ .The output was above 27.8 × 10^5/ml . Meanwhile the different factors, osmotic pressure stabilizer,medium,inoculation method and temperature, for protoplast reproduction of F. Velutipes were studled, the optimum condition of protoplast reproduction was obtained by spreading inoculation on RCM agar with 0.5mol/L KC1 at 25℃ ,The output was 0.5%.