Transgenic Populus tremula: a step-by-step protocol for its Agrobacterium-mediated transformation
作者： Tzvi Tzfira , Christian S. Jensen , Wangxia Wang , Amir Zuker , Basia Vinocur
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摘要: In recent years, Populus species have acquired an important place in basic and applied research of woody plants. The practical role world forestry their importance to as a woody-plant model led increasing interest tissue-culture molecular techniques, well the development transformation procedures for this genus. A simple technical procedure is described here step-by-step, first time, routine method transforming tremula using disarmed Agrobacterium tumefaciens hypervirulent strain. begins with inoculation stem explants bacterial suspension, followed by short period co-cultivation on highly regenerative medium. Transformed shoots are selected regeneration medium containing antibiotics presence inserted target genes checked rapid efficient PCR test. Selected transferred rooting medium, under same selection pressure, propagated via cuttings. plants can be hardened green-house within 4 months inoculation. has proven several gene constructs, Kan or Hyg, three different strains.
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