TRV-GFP: a modified Tobacco rattle virus vector for efficient and visualizable analysis of gene function.
TRV-GFP: a modified Tobacco rattle virus vector for efficient and visualizable analysis of gene function.
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TRVGFP：一种改良的烟草脆裂病毒载体，用于高效、可视化的基因功能分析
DOI：
10.1093/jxb/ert381
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发表时间：
2014-01
影响因子：
6.9
通讯作者：
Ma N
中科院分区：
文献类型：
--
作者：
Tian J;Pei H;Zhang S;Chen J;Chen W;Yang R;Meng Y;You J;Gao J;Ma N
我们开发了一种易于追踪的TRV载体，TRV2-GFP，通过在外壳蛋白上标记一个GFP。通过GFP监测可以有效识别trv2 -GFP感染植株。TRV2-GFP可用于许多植物的功能基因组学研究，特别是对非茄科植物，如玫瑰病毒诱导的基因沉默（VIGS）是植物基因功能表征的有用工具。不幸的是，烟草响尾蛇病毒（TRV）对一些非茄科植物的感染效率相对较低，这些植物如玫瑰（Rosa sp.）具有重要的经济价值。为了生成易于追溯的TRV载体，我们将绿色荧光蛋白（GFP）基因标记在原TRV2载体的外膜蛋白基因3 '端，并在本烟、拟南芥、玫瑰、草莓（Fragaria ananassa）、菊花（Dendranthema grandflorum）等几种植物中测试了修饰后的TRV - GFP载体的沉默效率。结果表明，TRV - gfp在各被试植株上的侵染效率与原TRV载体侵染效率相当。利用荧光显微镜和手持式紫外灯可方便地监测改良后的TRV病毒的传播情况。利用TRV-GFP沉默玫瑰插枝和幼苗的内源植物烯去饱和酶（PDS）基因后，75% ~ 80%的植株出现典型的光漂白表型，经紫外灯鉴定为GFP阳性。此外，代表TRV病毒浓度的GFP蛋白丰度与PDS基因的水平呈负相关，表明GFP可以作为靶基因沉默程度的指标。本研究为预测植物基因沉默阳性提供了一种可视化、高效的工具，对植物，特别是非茄科植物的基因功能研究具有重要价值。
We developed an easy-traceable TRV vector, TRV2-GFP, by tagging a GFP to the coat protein. TRV2-GFP-infected plants could be identified efficiently by GFP monitoring. TRV2-GFP is useful for functional genomics in many plants, especially for non-Solanaceae plants, like rose Virus-induced gene silencing (VIGS) is a useful tool for functional characterization of genes in plants. Unfortunately, the efficiency of infection by Tobacco rattle virus (TRV) is relatively low for some non-Solanaceae plants, which are economically important, such as rose (Rosa sp.). Here, to generate an easy traceable TRV vector, a green fluorescent protein (GFP) gene was tagged to the 3’ terminus of the coat protein gene in the original TRV2 vector, and the silencing efficiency of the modified TRV–GFP vector was tested in several plants, including Nicotiana benthamiana, Arabidopsis thaliana, rose, strawberry (Fragaria ananassa), and chrysanthemum (Dendranthema grandiflorum). The results showed that the efficiency of infection by TRV–GFP was equal to that of the original TRV vector in each tested plant. Spread of the modified TRV virus was easy to monitor by using fluorescent microscopy and a hand-held UV lamp. When TRV–GFP was used to silence the endogenous phytoene desaturase (PDS) gene in rose cuttings and seedlings, the typical photobleached phenotype was observed in 75–80% plants which were identified as GFP positive by UV lamp. In addition, the abundance of GFP protein, which represented the concentration of TRV virus, was proved to correlate negatively with the level of the PDS gene, suggesting that GFP could be used as an indicator of the degree of silencing of a target gene. Taken together, this work provides a visualizable and efficient tool to predict positive gene silencing plants, which is valuable for research into gene function in plants, especially for non-Solanaceae plants.
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影响因子：
4.4
作者：
Kim J;Park JH;Lim CJ;Lim JY;Ryu JY;Lee BW;Choi JP;Kim WB;Lee HY;Choi Y;Kim D;Hur CG;Kim S;Noh YS;Shin C;Kwon SY
通讯作者：
Kwon SY
影响因子：
5.1
作者：
Liu, Y;Nakayama, N;Dinesh-Kumar, SP
通讯作者：
Dinesh-Kumar, SP
影响因子：
7.2
作者：
Ratcliff, F;Martin-Hernandez, AM;Baulcombe, DC
通讯作者：
Baulcombe, DC
影响因子：
7.4
作者：
Burch-Smith, Tessa M.;Schiff, Michael;Dinesh-Kumar, S. P.
通讯作者：
Dinesh-Kumar, S. P.
影响因子：
4.4
作者：
Dubois A;Carrere S;Raymond O;Pouvreau B;Cottret L;Roccia A;Onesto JP;Sakr S;Atanassova R;Baudino S;Foucher F;Le Bris M;Gouzy J;Bendahmane M
通讯作者：
Bendahmane M