| 马铃薯疮痂病新致病种Streptomyces galilaeus CPS-2转化体系的建立 |
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| 引用本文： | 陈俊杰,赵伟全,于秀梅,刘大群.马铃薯疮痂病新致病种Streptomyces galilaeus CPS-2转化体系的建立[J].河南农业科学,2010(8). |
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| 作者姓名： | 陈俊杰 赵伟全 于秀梅 刘大群 |
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| 作者单位： | 1. 河北农业大学,植物保护学院,河北省农作物病虫害生物防治工程技术研究中心,河北,保定,071001
2. 河北农业大学,生命科学学院,河北,保定,071001 |
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| 基金项目： | 国家自然科学基金，国家863项目，现代农业产业技术体系建设专项资金资助项目 |
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| 摘 要： | 为建立马铃薯疮痂病新致病种Streptomyces galilaeus典型菌株CPS-2的转化体系,对该菌的原生质体制备和再生条件进行了筛选。结果表明,菌株CPS-2原生质体制备的最适条件为:TSB培养基一级培养24h,改良YEME培养基(含10%蔗糖和1.0%甘氨酸)二级培养60h,溶菌酶用量为3.0g/L,37℃酶解90 min,原生质体形成量可达4.4×109个/mL;在R2YE培养基上再生率可达22.71%。采用含25%PEG1000的Tbuffer介导转化质粒pIJ702,24h后覆盖含有硫链丝菌素的营养软琼脂,4d后可筛选到转化子,每微克质粒DNA可获得102~103个转化子。
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| 关 键 词： | 马铃薯疮痂病菌 原生质体 再生 转化 |
Construction the Protoplast Transformation System of Potato Common Scab Pathogen Streptomyces galilaeus CPS-2 |
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| CHEN Jun-jie,ZHAO Wei-quan,YU Xiu-mei,LIU Da-qun.Construction the Protoplast Transformation System of Potato Common Scab Pathogen Streptomyces galilaeus CPS-2[J].Journal of Henan Agricultural Sciences,2010(8). |
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| Authors: | CHEN Jun-jie ZHAO Wei-quan YU Xiu-mei LIU Da-qun |
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| Abstract: | The conditions of formation and regeneration of protoplast were screened in order to construct the protoplast transformation system of potato scab pathogen Streptomyces galilaeus(strain) CPS-2.The result showed that the optimum conditions for protoplast formation was as follows: shaking culture in TSB for 24h,transferring to YEME medium(containing 10% glucose and 1.0% glycine) and continuning shaking culture for 60h.The number of protoplasts per mL could up to 4.4×109 when the lysozyme concentration was 3.0g/L and incubated at 37℃ for 90min.The regeneration rate was 22.71% on R2YE medium.Plasmid pIJ702 was used to validated the protoplasts of strain CPS-2.Using T buffer containing 25% PEG 1000,102-103 transformants per μg plasmid DNA were obtained after four days. |
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| Keywords: | Potato scab pathogen Protoplast Regeneration Transformation |
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