荨麻青霉(Penicillium urticae)原生质体的制备与再生

Preparation and Regeneration of Protoplasts of Penicillium urticae

对荨麻青霉原生质体的高效制备以及再生方法进行了探索.结果显示荨麻青霉较为适合的原生质体制备及再生条件的组合:菌龄为28℃恒温下培养20～22h,DTT预处理0.5h,以7mg/mL的纤维素酶,7mg/mL蜗牛酶,5mg/mL溶菌酶为混合酶,0.6mol/L的NaCl作为渗透压稳定剂,28℃恒温酶解3.5h,PDA高渗双层培养基再生.该组合制备原生质体形成量达到1.59×107～1.89×107/mL,其再生率达到16.24%～19.25%.

The highly effective protoplast preparation and regeneration method of Penicillium urticae Bainier had been explored. The optimum osmotic pressure stabilizer was 0. 6 mol/L NaCl,and the optimum age of the mycelia was 20-22 hours cultivated in the temperature of 28 ℃. P. urticae Bainier hypha pretreated with DTT 0. 5 hour, hydrolysed 3.5 hours by enzyme mixture consisting of 7 mg/mL cellulase, 7 mg/mL snailase, and 5 mg/mL lysonzyme in the temperature at 28 ℃, and separated on the double-decked high infiltrate PDA culture medium plate. The results showed that the protoplast yield reached 1.59 ×10^7- 1.89×10^7/mL, and the regeneration rate of protoplast reached 16.24 % - 19.25 %, under aforementioned conditions.