Okadaic acid and trifluoperazine enhance <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation in eastern white pine |
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Authors: | Wei Tang Jinxing Lin Ronald J Newton |
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Institution: | (1) Department of Biology, Howell Science Complex, East Carolina University, Greenville, NC 27858-4353, USA;(2) Key Laboratory of Photosynthesis and Molecular Environment Physiology, Institute of Botany, The Chinese Academy of Sciences, Beijing, 100093, P.R. China |
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Abstract: | Mature zygotic embryos of recalcitrant Christmas tree species eastern white pine (Pinus strobus L.) were used as explants for Agrobacterium tumefaciens strain GV3101-mediated transformation using the uidA (β-Glucuronidase) gene as a reporter. Influence of the time of sonication and the concentrations of protein phosphatase inhibitor
(okadaic acid) and kinase inhibitor (trifluoperazine) on Agrobacterium-mediated transformation have been evaluated. A high transformation frequency was obtained after embryos were sonicated for
45–50 s, or treated with 1.5–2.0 μM okadaic acid or treated with 100–200 μM trifluoperazine, respectively. Protein phosphatase
and kinase inhibitors enhance Agrobacterium-mediated transformation in eastern white pine. A 2–3.5-fold higher rate of hygromycin-resistant callus was obtained with
an addition of 2 μM okadaic acid or 150 μM trifluoperazine or sonicated embryos for 45 s. Stable integration of the uidA gene in the plant genome of eastern white pine was confirmed by polymerase chain reaction (PCR), Southern and northern blot
analyses. These results demonstrated that a stable and enhanced transformation system has been established in eastern white
pine and this system would provide an opportunity to transfer economically important genes into this Christmas tree species.
Communicated by W. H. Wu |
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Keywords: | Agrobacterium tumefaciens Christmas tree species β -Glucuronidase Transgenic plants |
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