苹果腐烂病菌原生质体制备与再生条件优化

摘要

大量高质量且能再生的原生质体是真菌遗传转化、基因修饰的重要保证。本试验选用苹果腐烂病菌强致病力菌株SDAU11-175，从酶种类、菌丝年龄、酶解时间、渗透压稳定剂及再生培养基五个方面对原生质体制备及再生条件进行了优化。结果显示：获得苹果腐烂病菌原生质体最大产量的条件是菌丝年龄54 h、3％崩溃酶和3％裂解酶组合、0．7 mol/L NaCl作为渗透压稳定剂、28℃酶解3 h，所得原生质体在YPS培养基上再生效果最好，再生率可达42．21％。%Protoplasts with high quality and high regeneration rate are essential in genetic transformation and gene modification of fungi .Using highly pathogenic Valsa mali var.mali strain SDAU11-175 as research object , the protoplast preparation and regeneration conditions were optimized from five aspects of enzyme types, mycelia age, enzymolysis time, osmotic stabilizer and regeneration medium .The results showed that the conditions to obtain the maximum protoplast output were culturing mycelia for 54 hours and treating them with 3%driselase and 3% lysing enzymes for 3 hours at 28℃ using 0 .7 mol/L NaCl as osmotic stabilizer . The protoplasts regenerated best on YPS medium with the regeneration rate as 42 .21%.