Sonoda S, Nishiguchi M
National Institute of Agrobiological Resources, Kan-nondai 2-1-2, Tsukuba, Ibaraki 305-8602, Japan.
Plant J. 2000 Jan;21(1):1-8. doi: 10.1046/j.1365-313x.2000.00645.x.
Using the grafting procedure, we examined the transmission of post-transcriptional gene silencing (PTGS) in Nicotiana benthamiana which had been transformed with the coat protein gene, including the 3' non-translated region of the sweet potato feathery mottle potyvirus. Transmission of PTGS from silenced lines to non-silenced ones was bidirectional, but occurred efficiently from root stocks to scions. The level of transgene methylation in non-silenced scions grafted onto silenced root stocks was not increased. When grafted scions which had become silenced were removed from silenced root stocks and regrafted onto non-silenced or vector-transformed root stocks, PTGS was maintained. However, their progeny did not show PTGS. Previously we reported that our transgenic lines had different target specificities of PTGS for RNA degradation: one line recognized only the 3' part of the transgene mRNA while others involved the whole transgene mRNA (Sonoda et al. 1999, Phytopathology, 89, 385-391). Using these lines, we showed that target specificity of PTGS induced in non-silenced scions after grafting was determined by that in silenced root stocks. However, unexpectedly, target specificity of PTGS induced in silenced scions was not changed [corrected] by grafting onto silenced root stocks showing different target specificity, indicating that the second PTGS from silenced root stocks was not superimposed to silenced scions.
我们采用嫁接方法,研究了转录后基因沉默(PTGS)在已用包含甘薯羽状斑驳马铃薯Y病毒3'非翻译区的外壳蛋白基因转化的本氏烟草中的传递情况。PTGS从沉默株系向非沉默株系的传递是双向的,但从砧木到接穗的传递效率更高。嫁接到沉默砧木上的非沉默接穗中转基因甲基化水平并未升高。当已沉默的嫁接接穗从沉默砧木上移除并重新嫁接到非沉默或载体转化的砧木上时,PTGS得以维持。然而,它们的后代并未表现出PTGS。此前我们报道过,我们的转基因株系对RNA降解的PTGS具有不同的靶标特异性:一个株系仅识别转基因mRNA的3'部分,而其他株系涉及整个转基因mRNA(Sonoda等人,1999年,《植物病理学》,89卷,385 - 391页)。利用这些株系,我们发现嫁接后在非沉默接穗中诱导的PTGS的靶标特异性由沉默砧木中的靶标特异性决定。然而,出乎意料的是,嫁接到显示不同靶标特异性的沉默砧木上后,沉默接穗中诱导的PTGS的靶标特异性并未改变[修正],这表明来自沉默砧木的第二次PTGS并未叠加到沉默接穗上。