北京市农林科学院机构知识库

A viral satellite DNA vector-induced transcriptional gene silencing via DNA methylation of gene promoter in Nicotiana benthamiana

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文献类型：外文期刊

作者： Ju, Zheng ¹ ; Wang, Lei ¹ ; Cao, Dongyan ¹ ; Zuo, Jinhua ² ; Zhu, Hongliang ¹ ; Fu, Daqi ¹ ; Luo, Yunbo ¹ ; Zhu, Benzhong ¹ ;

作者机构： 1.China Agr Univ, Coll Food Sci & Nutr Engn, Beijing 100083, Peoples R China

2.Beijing Acad Agr & Forestry Sci, Natl Engn Res Ctr Vegetables, Beijing 100097, Peoples R China

关键词： Tomato yellow leaf curl China virus;Satellite DNA virus;DNA methylation;Virus-induced transcriptional gene silencing;Green fluorescent protein;Bisulfite sequencing

期刊名称：VIRUS RESEARCH （影响因子：3.303；五年影响因子：3.445 ）

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收录情况： SCI

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. (C) 2016 Elsevier B.V. All rights reserved.

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