中文摘要：病毒诱导的基因沉默（VIGS）已通过各种DNA或RNA病毒载体广泛应用于转录后植物功能基因组的研究。然而，病毒通过基因启动子DNA甲基化诱导的转录后基因沉默（VITGS）是通过几种植物RNA病毒载体实现的，这还未通过使用卫星DNA病毒载体实现。本研究探索将与中国番茄黄化曲叶病毒有关的病毒卫星DNA发展为VITGS载体。研究结果表明，病毒卫星DNA载体可作为一种有效的VITGS载体来研究植物基因组DNA甲基化。
外文摘要：Virus-induced gene silencing (VIGS) has been widely used for plant functional genomics study at the post-transcriptional level using various DNA or RNA viral vectors. However, while virus-induced transcriptional gene silencing (VITGS) via DNA methylation of gene promoter was achieved using several plant RNA viral vectors, it has not yet been done using a satellite DNA viral vector. In this study, a viral satellite DNA associated with tomato yellow leaf curl China virus (TYLCCNV), which has been modified as a VIGS vector in previous research, was developed as a VITGS vector. Firstly, the viral satellite DNA VIGS vector was further optimized to a more convenient p1.7A+ 2m beta vector with high silencing efficiency of the phytoene desaturase (PDS) gene in Nicotiana benthamiana plants. Secondly, the constructed VITGS vector (TYLCCNV:35S), which carried a portion of the cauliflower mosaic virus 35S promoter, could successfully induce heritable transcriptional gene silencing (TGS) of the green fluorescent protein (GFP) gene in the 35S-GFP transgenic N.benthamiana line 16c plants. Moreover, bisulfite sequencing results revealed higher methylated cytosine residues at CG, CHG and CHH sites of the 35S promoter sequence in TYLCCNV:35S-inoculated plants than in TYLCCNV-inoculated line 16c plants (control). Overall, these results demonstrated that the viral satellite DNA vector could be used as an effective VITGS vector to study DNA methylation in plant genomes.
外文关键词：Tomato yellow leaf curl China virus；Satellite DNA virus；DNA methylation；Virus-induced transcriptional gene silencing；Green fluorescent protein；Bisulfite sequencing
作者：Ju, Zheng；Wang, Lei；Cao, Dongyan；等
作者单位：中国农业大学
期刊名称：VIRUS RESEARCH
期刊影响因子：2.526
出版年份：2016
出版刊次：9
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