| 苹果腐烂病菌原生质体制备与再生条件优化 |
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| 引用本文： | 陈亮,伦莹莹,孙庚午,赵迎彤,何邦令,王洪凯,刘会香.苹果腐烂病菌原生质体制备与再生条件优化[J].山东农业科学,2014(8):109-112. |
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| 作者姓名： | 陈亮 伦莹莹 孙庚午 赵迎彤 何邦令 王洪凯 刘会香 |
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| 作者单位： | 山东农业大学植物保护学院;浙江大学生物技术研究所; |
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| 基金项目： | 国家科技部基础平台项目(2009FY210100);世界银行贷款山东生态造林项目(SEAP-JC-2);浙江省自然科学基金项目(Y3110028) |
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| 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%。
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| 关 键 词： | 苹果腐烂病菌 原生质体 制备 再生 |
Optimization of Protoplast Preparation and Regeneration Conditions of Valsa mali var.mali |
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| Institution: | Chen Liang, Lun Yingying, Sun Gengwu, Zhao Yingtong, He Bangling, Wang Hongkai, Liu Huixiang ( 1. College of Plant Protection, Shandong Agricultural University, Taian 271018, China ; 2. Biotechnology Institute, Zhejiang University, Hangzhou 310058, China) |
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| Abstract: | 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%. |
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| Keywords: | Valsa mali var mali Protoplast Preparation Regeneration |
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