赤霉菌原生质体外源基因高效转化方法研究  

A high efficiency method for genetic transformation of Gibberella fujikuroi by electroporation

作　　者：朱廷恒[1] 易佳炜 王渭霞[2] 李睿[3] 汪琨[1] 崔志峰[1] 

机构地区：[1]浙江工业大学生物与环境工程学院,浙江杭州310032 [2]中国水稻研究所,浙江杭州310006 [3]武汉工业学院生物与制药工程学院,湖北武汉430023

出　　处：《浙江工业大学学报》2013年第5期482-485,共4页Journal of Zhejiang University of Technology

基　　金：国家自然科学基金资助项目(31272002)

摘　　要：赤霉素(gibberellic acid,GA)是一种非常重要的植物生长调节剂,具有促进植物种子萌发和生长等功效.生产上用的赤霉素主要通过水稻赤霉病菌藤仓赤霉(Gibberella fujikuroi)工业发酵而来.赤霉菌工业生产菌株不产生孢子,不易进行以孢子为受体的外源基因转化.本研究建立了工业赤霉菌原生质体制备以及以原生质体为受体、利用电击高效转化外源基因的方法.结果发现崩溃酶(Dryslase)和溶壁酶(Lysing enzyme)以3:7的质量比混合时对赤霉菌原生质体的制备效果最好.电压为0.8～1.0kV的范围内原生质体的转化率最高,1μg DNA可以得到18个转化子.提供的方法不仅可以用于藤仓赤霉工业菌株的遗传改良,还可以用于水稻赤霉病菌分子生物学研究.Gibberellic acid （GA） is a very important plant hormone that can improve seed germination and plant growth. Industrially produced GA through fermentation with the fungus Gibberella fujikuroi is mainly used. As the industrial strain of this fungus does not produce spores, DNA transformation with spores as receptor was limited. A method for protoplast preparation and DNA transformation of G. fujikuroi industrial strain by electroporation was provided here. The result showed that the mixture of dryslase and lysing enzyme at an mass ratio of 3：7 had a good effect for protoplast preparation. Electroporation with the parameter value of the voltage between 0.8 and 1.0 kV got the highest transformation rate, 18 transformants per gg DNA. This method can be applied not only in genetic improvement of G. fujikuroi industrial strain,but also in study of molecular biology within this rice pathogen.

关 键 词：藤仓赤霉 原生质体 电转化 

分 类 号：Q933[生物学—微生物学]