A Virus-Induced Assay for Functional Dissection and Analysis of Monocot and Dicot Flowering Time Genes
A Virus-Induced Assay for Functional Dissection and Analysis of Monocot and Dicot Flowering Time Genes
复制标题
用于单子叶植物和双子叶植物开花时间基因功能解剖和分析的病毒诱导测定
DOI：
10.1104/pp.17.00392
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发表时间：
2017-06-01
期刊：
影响因子：
7.4
通讯作者：
Hong, Yiguo
中科院分区：
文献类型：
--
作者：
Qin, Cheng;Chen, Weiwei;Hong, Yiguo
病毒诱导开花（VIF）是利用病毒载体表达开花位点T（FT）来诱导植物开花。这种方法最近因其在加速作物和木本果树育种中的实际应用而引起了广泛的兴趣。然而，深入了解VIF及其作为解剖成花蛋白的有力工具的潜力仍有待阐明。在这里，我们描述了马铃薯X病毒（PVX）为基础的VIF在短日照烟草品种马里兰州猛犸的机制和进一步的应用。异位交付拟南芥（拟南芥）AtFT PVX/AtFT没有诱导内源性FT直系同源物NtFT 4的表达，但是，它足以触发开花的马里兰州猛犸植物生长在无诱导长日照条件下。受感染的烟草植物没有出现系统性症状，并且基于PVX的VIF没有引起跨代开花。我们表明，PVX为基础的VIF是一个更快速的方法来检查的影响，单个氨基酸突变AtFT的花诱导比每个突变的转基因拟南芥系。我们还使用基于PVX的VIF来证明，在AtFT的N或C末端添加His或FLAG标签可以影响其成花活性，并且该系统可以应用于检测来自异源物种的FT基因的功能，包括番茄（Solanum lycopersicum）SFT和水稻（Oryza sativa）Hd 3a。因此，基于PVX的VIF代表了一个简单而有效的系统，以确定个别的氨基酸是必不可少的FT介导的成花诱导和测试的能力，单子叶和双子叶FT基因和FT融合蛋白诱导开花。
Virus-induced flowering (VIF) uses virus vectors to express Flowering Locus T (FT) to induce flowering in plants. This approach has recently attracted wide interest for its practical applications in accelerating breeding in crops and woody fruit trees. However, the insight into VIF and its potential as a powerful tool for dissecting florigenic proteins remained to be elucidated. Here, we describe the mechanism and further applications of Potato virus X (PVX)-based VIF in the short-day Nicotiana tabacum cultivar Maryland Mammoth. Ectopic delivery of Arabidopsis (Arabidopsis thaliana) AtFT by PVX/AtFT did not induce the expression of the endogenous FT ortholog NtFT4; however, it was sufficient to trigger flowering in Maryland Mammoth plants grown under noninductive long-day conditions. Infected tobacco plants developed no systemic symptoms, and the PVX-based VIF did not cause transgenerational flowering. We showed that the PVX-based VIF is a much more rapid method to examine the impacts of single amino acid mutations on AtFT for floral induction than making individual transgenic Arabidopsis lines for each mutation. We also used the PVX-based VIF to demonstrate that adding a His-or FLAG-tag to the N or C terminus of AtFT could affect its florigenic activity and that this system can be applied to assay the function of FT genes from heterologous species, including tomato (Solanum lycopersicum) SFT and rice (Oryza sativa) Hd3a. Thus, the PVX-based VIF represents a simple and efficient system to identify individual amino acids that are essential for FT-mediated floral induction and to test the ability of mono-and dicotyledonous FT genes and FT fusion proteins to induce flowering.