Establishment of the straightforward electro-transformation system for Phytophthora infestans and its comparison with the improved PEG/CaCl2 transformation
Establishment of the straightforward electro-transformation system for Phytophthora infestans and its comparison with the improved PEG/CaCl2 transformation
复制标题
致病疫霉直接电转化体系的建立及其与改良PEG/CaCl2转化体系的比较
DOI：
10.1016/j.mimet.2015.03.013
复制
发表时间：
2015-05-01
影响因子：
2.2
通讯作者：
Liu, Huirong
中科院分区：
文献类型：
--
作者：
Dong, Lei;Zhu, Xiangyuan;Liu, Huirong
致病疫霉是马铃薯上最具毁灭性的病原菌。致病疫霉的遗传转化技术是致病疫霉分子生物学研究的难点之一。本研究以马铃薯晚疫病菌（P. infestans）为研究对象，以绿色荧光蛋白表达载体为载体，建立了马铃薯晚疫病菌的直接电转化体系，并与改良的PEG/CaCl 2介导的原生质体转化体系进行了比较。结果表明，直接电转化法在致病疫霉中可行，每1.10 × 10（6）个游动孢子可获得32个阳性转化子。转化子/μ g载体DNA为1.08。直接电转化的转化效率比PEG/CaCl 2介导的原生质体转化的转化效率高约2倍（每约1.05 × 10（6）原生质体17个阳性转化子，每μ g载体DNA 0.58个转化子）。与改良PEG/CaCl 2转化法相比，直接电转化法更简单，从收集材料到获得抗性转化子所需的起始材料和操作时间更少。本研究为进一步开展马铃薯晚疫病菌的生物学研究奠定了基础。(C)2015 Elsevier B. V.版权所有。
Phytophthora infestans is the most devastating pathogen of potato. For the biology study of P. infestans at the molecular level, one of the difficulties is the technique for the genetic transformation. In this study, the straightforward electro-transformation system was established for P. infestans with a green fluorescent protein expression vector and compared with the improved PEG/CaCl2 mediated protoplast transformation system. The results showed that the straightforward electro-transformation could work in P. infestans and 32 positive transformants were obtained per about 1.10 x 10(6) zoospores. The transformants per mu g of vector DNA were 1.08. The transformation efficiency of the straightforward electro-transformation was approximately 2 times higher than that of the improved PEG/CaCl2 mediated protoplast transformation (17 positive transformants per about 1.05 x 10(6) protoplasts, 0.58 transformants per mu g of vector DNA) according to the reported procedures. Furthermore, compared with the improved PEG/CaCl2 transformation, the straightforward electroporation is simpler and requires less starting materials and operating time from collecting material to obtaining the resistant transformants. Our work will lay a foundation for the biology study of P. infestans in the future. (C) 2015 Elsevier B.V. All rights reserved.