An improved transformation system for Lombardy poplar (Populus nigra var. italica)
An improved transformation system for Lombardy poplar (Populus nigra var. italica)
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DOI：
10.1007/s10310-006-0203-1
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发表时间：
2006-06-01
影响因子：
1.5
通讯作者：
Shinohara, Kenji
中科院分区：
文献类型：
--
作者：
Nishiguchi, Mitsuru;Yoshida, Kazumasa;Shinohara, Kenji
本文报道了一种改良的伦巴第杨转化体系。意大利语)。构建了含有11个独特酶切位点和正常新霉素磷酸转移酶II(NPTII)基因的双元载体pBF2，用于转化伦巴第杨。将根癌农杆菌EHA105与含有NPTII基因和增强型绿色荧光蛋白基因的双元载体共培养后，直接再生了转基因不定芽。通过荧光显微镜、免疫印迹和Southern印迹分析，以及对卡那霉素和遗传素的抗性，证实了转化成功。这种转化体系比我们以前的方法再生转基因芽所需的时间更短。当外植体在含有二硫苏糖醇的培养基上培养时，转化率提高到约20%。
We report an improved transformation system for Lombardy poplar (Populus nigra var. italica). A new binary vector, pBF2, with 11 unique restriction enzyme sites and the normal neomycin phosphotransferase II (NPTII) gene was constructed for the transformation of Lombardy poplar. Genetically transformed adventitious shoots were directly regenerated after cocultivation of stem segments with Agrobacterium tumefaciens EHA105 that harbored a binary vector with genes for the NPTII and enhanced green fluorescent protein. Successful transformation was confirmed by fluorescence microscopy, immunoblotting and Southern blotting analyses, and resistance to kanamycin and geneticin. This transformation system requires less time than our previous method for the regeneration of transgenic shoots. When explants were incubated on a smedium containing dithiothreitol, the transformation frequency increased to approximately 20%.