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Genetic transformation of golden pothos ( Epipremnum aureum ) mediated by Agrobacterium tumefaciens

机译:根癌农杆菌介导的金马铃薯(金黄色葡萄球菌)的遗传转化

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摘要

To establish a procedure for Agrobacterium tumefaciens-mediated transformation of golden pothos (Epipremnum aureum) plants, the effects of selection antibiotics and the preculture period of stem explants before A. tumefaciens infection were examined. Explants were co-cultivated with A. tumefaciens EHA105, harboring the plasmid pGWB2/cGUS, on a somatic embryo-inducing medium supplemented with acetosyringone. Resulting transgenic somatic embryos were screened on an antibiotic selection medium, and the transgenic pothos plants were regenerated on a germination medium. Hygromycin was the optimum selection antibiotic tested. The preculture period significantly affected the transformation efficiency, with explants precultured for one-day showing the best efficiency (5–30%). Both transformed hygromycin-resistant embryos and regenerated plants showed β-glucuronidase activity. Southern blot analysis confirmed transgene integration into the pothos genome. This reproducible transformation system for golden pothos may enable the molecular breeding of this very common indoor plant.
机译:为了建立根癌农杆菌介导的金马铃薯(Epipremnum aureum)植物转化的程序,研究了选择抗生素的作用和根癌农杆菌感染前茎外植体的预培养期。将外植体与携带质粒pGWB2 / cGUS的根癌农杆菌EHA105在补充乙酰丁香酮的体细胞胚诱导培养基上共培养。在抗生素选择培养基上筛选得到的转基因体细胞胚,并在萌发培养基上再生转基因马铃薯。潮霉素是测试的最佳选择抗生素。预培养期显着影响转化效率,预培养一天的外植体显示出最高的转化效率(5–30%)。转化的潮霉素抗性胚和再生植物均显示β-葡萄糖醛酸苷酶活性。 Southern印迹分析证实了转基因整合到pothos基因组中。这种用于金色pothos的可重现的转化系统可以使这种非常常见的室内植物进行分子育种。

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