桉树焦枯病菌原生质体制备条件的优化  

Optimization on preparation condition of protoplast from Calonectria pseudoreteaudii

作　　者：叶小真[1,2] 郭文硕[1] 冯丽贞[1] 金亚杰[1] 杨新新[2] 刘宏毅[1] 连鑫坤 李慧敏[1] 

机构地区：[1]福建农林大学林学院 [2]福建农林大学金山学院,福建福州350002

出　　处：《福建农林大学学报（自然科学版）》2016年第2期141-145,共5页Journal of Fujian Agriculture and Forestry University:Natural Science Edition

基　　金：福建省教育厅项目(JA13372);福建省财政厅社会公益项目(K8112004A);福建省财政厅项目(Scz10039)

摘　　要：以桉树焦枯病强致病菌株Calonectria pseudoreteaudii YA51为材料,对其原生质体生成条件进行优化.结果表明,收集1 g(湿重)培养24 h的菌丝,以0.8 mol·L^(-1)Na Cl为渗透压稳定剂,置于10 m L含20 mg·m L^(-1)崩溃酶和30 mg·m L^(-1)裂解酶的混合酶液中,在30℃、100 r·min^(-1)条件下裂解4 h,原生质体生成量可达3.72×107个·m L^(-1),在SR培养基上的再生率为32.33%.To effectively remove cell wall without breaking osmotic pressure equilibrium,protoplast preparation conditions of virulent strain Calonectria pseudoreteaudii YA51 were studied. The results showed that 10 m L enzyme mixture which included 20 mg · m L^-1driselase and 30 mg·m L^-1lysing enzyme had the best effect on cell wall degradation of C. pseudoreteaudii YA51. By using 0. 8 mol·L^-1Na Cl as buffer,1 g mycelium cultured in PDB for 24 hours was hydrolyzed in 10 m L enzymes mixture at 30 ℃ for 4 hours at 100 r·min^-1. The protoplast yield was to 3. 72 × 107cell·m L^-1. Morever,32. 33% protoplasts could regenerate in SR medium.

关 键 词：桉树焦枯病菌 原生质体 生成量 条件优化 

分 类 号：S763.15[农业科学—森林保护学]