Truve E, Kelve M, Aaspôllu A, Kuusksalu A, Seppänen P, Saarma M
Institute of Chemical Physics and Biophysics, Estonian Academy of Sciences, Tallinn, Estonia.
Arch Virol Suppl. 1994;9:41-50. doi: 10.1007/978-3-7091-9326-6_5.
We investigated the possibility of reconstructing the 2'-5' oligoadenylate (2-5A) pathway into the plant kingdom to achieve multiple virus resistance. Differently phosphorylated 2-5A trimers and tetramers inhibited TMV RNA translation in cell-free systems. In wheat germ extracts the most potent inhibitors were nonphosphorylated forms of 2-5A. Triphosphorylated forms of 2-5A were deposphorylated and hydrolysed in plant extracts. Since we could not detect homologous DNA to mammalian 2-5A synthetase cDNA in tobacco or potato, we cloned rat 2-5A synthetase cDNA and transformed it by the Agrobacterium-mediated mechanism into tobacco and potato. Transformed tobacco plants were resistant to PVS infection and propagation of PVX was reduced. In transgenic potatoes tolerance to PVX and, in one transgenic clone, also to PVY was observed.
我们研究了将2'-5'寡腺苷酸（2-5A）途径重建到植物界以实现多种病毒抗性的可能性。不同磷酸化的2-5A三聚体和四聚体在无细胞系统中抑制烟草花叶病毒（TMV）RNA翻译。在小麦胚芽提取物中，最有效的抑制剂是2-5A的非磷酸化形式。2-5A的三磷酸化形式在植物提取物中被去磷酸化并水解。由于我们在烟草或马铃薯中未检测到与哺乳动物2-5A合成酶cDNA同源的DNA，我们克隆了大鼠2-5A合成酶cDNA，并通过农杆菌介导的机制将其转化到烟草和马铃薯中。转化后的烟草植株对马铃薯纺锤块茎类病毒（PVS）感染具有抗性，并且马铃薯X病毒（PVX）的繁殖减少。在转基因马铃薯中观察到对PVX的耐受性，并且在一个转基因克隆中还观察到对马铃薯Y病毒（PVY）的耐受性。
