TRANSFORMATION OF BRASSICA-NAPUS AND BRASSICA-OLERACEA USING AGROBACTERIUM-TUMEFACIENS AND THE EXPRESSION OF THE BAR AND NEO GENES IN THE TRANSGENIC PLANTS
TRANSFORMATION OF BRASSICA-NAPUS AND BRASSICA-OLERACEA USING AGROBACTERIUM-TUMEFACIENS AND THE EXPRESSION OF THE BAR AND NEO GENES IN THE TRANSGENIC PLANTS
复制标题
DOI：
10.1104/pp.91.2.694
复制
发表时间：
1989-10-01
期刊：
影响因子：
7.4
通讯作者：
TENNING, P
中科院分区：
文献类型：
--
作者：
DEBLOCK, M;DEBROUWER, D;TENNING, P
基于neoorbaras选择标记基因，建立了一种高效的、基因型无关的油菜和甘蓝转化方法。用含有嵌合neo和bargene的农杆菌菌株感染甘蓝型油菜和甘蓝型油菜的下胚轴外植体。AgNO 3的使用是在选择性条件下高效再生芽的先决条件。通过降低培养基的水势、降低组织培养容器中的相对湿度和降低细胞分裂素浓度来避免玻璃化。以这种方式，在9至12周内以30%的效率获得生根的转化芽。Southern印迹和S1-后代的遗传分析表明，转化体平均含有1至3个拷贝的嵌合基因。在独立转化体中观察到嵌合基因的广泛表达水平。高达25%的转化体没有表现出可检测的草丁膦乙酰转移酶或新霉素磷酸转移酶II酶活性，尽管Southern印迹证明这些植物确实被转化。
An efficient and largely genotype-independent transformation method forBrassica napusandBrassica oleraceawas established based onneoorbaras selectable marker genes. Hypocotyl explants ofBrassica napusandBrassica oleraceacultivars were infected withAgrobacteriumstrains containing chimericneoandbargenes. The use of AgNO3was a prerequisite for efficient shoot regeneration under selective conditions. Vitrification was avoided by decreasing the water potential of the medium, by decreasing the relative humidity in the tissue culture vessel, and by lowering the cytokinin concentration. In this way, rooted transformed shoots were obtained with a 30% efficiency in 9 to 12 weeks. Southern blottings and genetic analysis of S1-progeny showed that the transformants contained on average between one and three copies of the chimeric genes. A wide range of expression levels of the chimeric genes was observed among independent transformants. Up to 25% of the transformants showed no detectable phosphinotricin acetyltransferase or neomycin phosphotransferase II enzyme activities although Southern blottings demonstrated that these plants were indeed transformed.