Transformation of plant young proembryos by electroporation |
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Authors: | Shitao Li Shenghua Wang Hongyuan Yang |
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Institution: | (1) Research Center for Developmental Biology, Wuhan University, 430072 Wuhan, China |
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Abstract: | It is first reported that plant young proembryos expressed exogenous reporter genes by electroporation. Young proembryos with
8–32 cells and globular proembryos with 250–400 cells could be isolated by enzymatic maceration combined with microdissection.
After electroporation withGUS orGFP genes, the proembryos were cultured for 1–2 d in KM8p medium. At the field strength of electroporation 500–1500 V/cm, blue
reaction of GUS or green fluorescence of GFP could be observed in the proembryos. The highest transient expression frequency
of young proembryos (2.2%) was obtained at the field strength of 750 V/cm, whereas the highest frequency of globular proembryos
(5.9%) was obtained at the field strength of 1 250 V/cm. Taking the proportion of transformed cells in the whole cells of
proembryos as efficient transformation frequency, the efficient transformation frequency of the young proembryos was 7 times
that of the globular proembryos. |
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Keywords: | electroporation GUS GFP proembryo Nicotiana tabacum |
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