Genetic Transformation Of Date Palm Via Microprojectile Bombardment

DATE PALM BIOTECHNOLOGY PROTOCOLS VOL I: TISSUE CULTURE APPLICATIONS(2017)

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摘要
Efficient protocols for date palm embryogenic callus and somatic embryo transformation with uidA gene are described in this chapter. The embryogenic callus transformation procedure is 1.6 mu m gold particle size coated with 2.5 mu g DNA (pAct1-D plasmid), 1100 psi helium pressure, 9 cm target distance, 26 inHg vacuum pressure, 3mmdistance between the rupture disk and macrocarrier, and osmotic pretreatment with 0.4 M mannitol followed by 60 min air desiccation. The somatic embryo transformation procedure is 0.6 mu m gold particle size coated with 2.5 mu g DNA (pAct1-D plasmid), 1350 psi helium pressure, 6 cm target distance, 28 inHg vacuum pressure, 3 mm distance between the rupture disk and macrocarrier, and osmotic pretreatment with 0.4 M mannitol followed by 60 min air desiccation. Protocols for analysis of the transgenic plantlets have also been described.
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关键词
Particle bombardment, Particle delivery, Transformation, Transgenic
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