Schmidt Janice F, Moore Maria D, Pelcher Lawrence E, Covello Patrick S
Plant Biotechnology Institute, 110 Gymnasium Place, S7N OW9 Saskatoon, SK, Canada.
Plant Cell Rep. 2007 Sep;26(9):1547-54. doi: 10.1007/s00299-007-0369-9. Epub 2007 May 22.
A highly efficient and convenient method for the Agrobacterium rhizogenes-dependent production of transformed roots of Saponaria vaccaria L. (Caryophyllaceae) is described. The parameters tested and optimized include S. vaccaria cultivar, explant type, Agrobacterium rhizogenes strain and culture conditions. For cotransformation using additional recombinant T-DNA-containing A. rhizogenes strains, use of neomycin phosphotransferase and enhanced green fluorescent protein genes as selectable markers were tested alone and in combination. Optimal results, yielding a minimum of one transformed root per explant, were obtained using the cultivar Pink Beauty, the A. rhizogenes strain LBA9402 and internode explants precultured on a phytohormone mixture. Selection of cotransformed roots by observation of enhanced green fluorescent protein fluorescence alone was highly effective and convenient.
本文描述了一种高效便捷的方法，用于发根农杆菌介导的肥皂草（石竹科）转化根的生产。所测试和优化的参数包括肥皂草品种、外植体类型、发根农杆菌菌株和培养条件。对于使用额外含重组T-DNA的发根农杆菌菌株进行共转化，单独及组合测试了使用新霉素磷酸转移酶和增强型绿色荧光蛋白基因作为选择标记。使用“粉红佳人”品种、发根农杆菌菌株LBA9402以及在植物激素混合物上预培养的节间外植体，获得了最佳结果，每个外植体至少产生一条转化根。仅通过观察增强型绿色荧光蛋白荧光来选择共转化根非常有效且便捷。
