title: Establishment of the straightforward electro-transformation system for Phytophthora infestans and its comparison with the improved PEG/CaCl₂ transformation.
abstract: 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 1.10×10(6) zoospores. The transformants per μg 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 1.05×10(6) protoplasts, 0.58 transformants per μg 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.
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Life Sciences, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot 010018, Inner Mongolia, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Dong
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Life Sciences, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot 010018, Inner Mongolia, China.
authors.forename: Xiangyuan
authors.initials: X
authors.lastname: Zhu
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Life Sciences, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot 010018, Inner Mongolia, China.
authors.forename: Haichen
authors.initials: H
authors.lastname: Cui
authors.affiliation: Department of Applied Molecular Biosciences, Graduate School of Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8601, Japan.
authors.forename: Makoto
authors.initials: M
authors.lastname: Ojika
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Life Sciences, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot 010018, Inner Mongolia, China.
authors.forename: Ruigang
authors.initials: R
authors.lastname: Wang
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Life Sciences, Inner Mongolia Agricultural University, 306 Zhaowuda Road, Hohhot 010018, Inner Mongolia, China. Electronic address: huirong_liu@imau.edu.cn.
authors.forename: Huirong
authors.initials: H
authors.lastname: Liu
mesh_terms: D018274:Electroporation / Q000379:methods*; D018014:Gene Transfer Techniques; D055750:Phytophthora infestans / Q000235:genetics*; D014170:Transformation, Genetic
keywords: Electroporation; Phytophthora; Protoplast; Transformation; Zoospore