title: A rapid, highly efficient and economical method of Agrobacterium-mediated in planta transient transformation in living onion epidermis.
abstract: Transient transformation is simpler, more efficient and economical in analyzing protein subcellular localization than stable transformation. Fluorescent fusion proteins were often used in transient transformation to follow the in vivo behavior of proteins. Onion epidermis, which has large, living and transparent cells in a monolayer, is suitable to visualize fluorescent fusion proteins. The often used transient transformation methods included particle bombardment, protoplast transfection and Agrobacterium-mediated transformation. Particle bombardment in onion epidermis was successfully established, however, it was expensive, biolistic equipment dependent and with low transformation efficiency. We developed a highly efficient in planta transient transformation method in onion epidermis by using a special agroinfiltration method, which could be fulfilled within 5 days from the pretreatment of onion bulb to the best time-point for analyzing gene expression. The transformation conditions were optimized to achieve 43.87% transformation efficiency in living onion epidermis. The developed method has advantages in cost, time-consuming, equipment dependency and transformation efficiency in contrast with those methods of particle bombardment in onion epidermal cells, protoplast transfection and Agrobacterium-mediated transient transformation in leaf epidermal cells of other plants. It will facilitate the analysis of protein subcellular localization on a large scale.
authors.affiliation: Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Kedong
authors.initials: K
authors.lastname: Xu
authors.affiliation: Department of Life Science, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Xiaohui
authors.initials: X
authors.lastname: Huang
authors.affiliation: Department of Life Science, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Manman
authors.initials: M
authors.lastname: Wu
authors.affiliation: Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China ; College of Life Science, Henan Agricultural University, Zhengzhou, People's Republic of China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Life Science, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Yunxia
authors.initials: Y
authors.lastname: Chang
authors.affiliation: Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Kun
authors.initials: K
authors.lastname: Liu
authors.affiliation: Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Ju
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Fuli
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Life Science, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Liming
authors.initials: L
authors.lastname: Yi
authors.affiliation: Department of Life Science, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Li
authors.affiliation: Department of Life Science, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Ruiyue
authors.initials: R
authors.lastname: Wang
authors.affiliation: Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Guangxuan
authors.initials: G
authors.lastname: Tan
authors.affiliation: Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China.
authors.forename: Chengwei
authors.initials: C
authors.lastname: Li
mesh_terms: D060054:Agrobacterium / Q000378:metabolism*; D017360:Arabidopsis / Q000382:microbiology; D019470:Biolistics; D005821:Genetic Techniques / Q000191:economics*; D019697:Onions / Q000235:genetics* / Q000382:microbiology*; D019441:Plant Epidermis / Q000166:cytology / Q000382:microbiology*; D030821:Plants, Genetically Modified; D015203:Reproducibility of Results; D013347:Subcellular Fractions / Q000378:metabolism; D013997:Time Factors; D014026:Nicotiana / Q000382:microbiology; D014170:Transformation, Genetic