丝状真菌AL18的原生质体制备和再生条件的优化  

Optimization of Preparation and Regeneration Condition of Protoplast from Filamentous Fungi AL18

作　　者：卢明锋[1] 张月杰[1] 

机构地区：[1]山东理工大学生命科学学院,山东淄博255091

出　　处：《生物技术》2012年第3期86-90,共5页Biotechnology

基　　金：国家重点基础研究计划项目("绿色化学农药先导结构及作用靶标的发现与研究";2003CB114400);云南省自然科学基金项目("新型光敏抗肿瘤药物先导结构的发现";2005C0078M)资助

摘　　要：目的:为了建立起产苝醌类光敏剂丝状真菌AL18原生质体制备和再生体系。方法:采用单因素实验法研究了预处理方式、渗透压稳定剂和酶解条件对原生质体制备率和再生率的影响。结果:原生质体制备及再生的最佳条件是用0.3%的β-巯基乙醇预处理15 min,酶解液以0.6 mol/L的MgSO4·7H2O作为渗透压稳定剂,0.02 mol/L的磷酸盐缓冲液pH值为5.8,纤维素酶:蜗牛酶=2:3,酶的总浓度为15mg/mL,36℃酶解2h;以0.6mol/L的蔗糖作为再生培养基的渗透压稳定剂。结论:原生质体的制备率和再生率可分别达到1.42×107/mL和3.2%。Objective:In order to establish preparation and regeneration system of protoplast from filamentous fungi AL18 producing perylenequinones photosensitizer.Method:Pretreatment,osmotic pressure stabilizer and enzymes digesting conditions were investigated by single factor design method.Result:The optimal condition is pretreatment mycelia with 0.3%β-mercaptoethanol for 15min,using 0.6 mol/L MgSO4·7H2O as osmotic pressure stabilizer,cellulose:snailase = 2:3 enzymes mixture which total concentration is 15mg/mL,enzymes digesting at 36℃ for 2 hours,applying 0.6 mol/L sucrose for osmotic pressure stabilizer in regeneration medium.Conclusion:The yield and regenerating rate of protoplast were achieved 1.42×106/mL,3.2% respectively.

关 键 词：苝醌类 光敏剂 原生质体 制备 再生 

分 类 号：Q933[生物学—微生物学]