摘要
报道一种经济高效的禾谷镰孢菌原生质体的转化方法。制备禾谷镰孢菌原生质体的最佳条件为每毫升2% Drislase和2% Snailase混合酶液中加入0.12g幼殖体,于30℃酶解1.5h。其转化效率约为40–50个转化子/μg片段DNA;转化子的PCR检测结果表明,潮霉素B基因已插入禾谷镰孢菌的基因组中。建立的禾谷镰孢菌原生质体转化系统具有经济和高效的特点,为禾谷镰孢菌基因功能的研究提供了必要的技术支持。
Protoplast transformation for Fusarium graminearum is an important technique for establishing mutation library and studying functional genome. In the current study, the optimum condition for preparing protoplasts was set up by using one milliliter enzyme mixture of 2% driselase and 2% snailase to digest the 0.12g germlings at 30℃ for 1.5 hours. PCR detection indicated that the transformation efficiency was 40-50 transformants per microgram of DNA and the fragment hph was inserted into the genome of E graminearum. The new system of protoplast transformation for F. graminearum was very economic and efficient. It provides an essential technology for establishing mutation library and studying the functional genome ofF. graminearum .
出处
《菌物学报》
CAS
CSCD
北大核心
2013年第5期891-898,共8页
Mycosystema
基金
Supported by National Natural Science Foundation of China(No.31272065,31171880 and 31201543)
National High-Technology Research and Development Program of China(No.2012AA101502)
the National Science and Technology Support Program(No.2012BAD19B01)
the Youth Science and Technology Innovation Fund of Nanjing Agricultural University(No.KJ2012005)
关键词
幼殖体
崩溃酶
蜗牛酶
germlings, driselase, snailase
