Virus-induced gene silencing for comparative functional studies in Gladiolus hybridus
Virus-induced gene silencing for comparative functional studies in Gladiolus hybridus
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DOI：
10.1007/s00299-013-1530-2
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发表时间：
2014-02
影响因子：
6.2
通讯作者：
Xiong-hui Zhong;Xueyuan Yuan;Ze Wu;M. A. Khan;Jin Chen;Xiao-xin Li;Benhe Gong;Yang Zhao;Jian W
中科院分区：
文献类型：
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作者：
Xiong-hui Zhong;Xueyuan Yuan;Ze Wu;M. A. Khan;Jin Chen;Xiao-xin Li;Benhe Gong;Yang Zhao;Jian W
由于缺乏一种高效稳定的遗传转化方法，唐菖蒲基因的功能分析一直是不切实际的。然而，病毒诱导的基因沉默(VIGS)在一些难以通过其他方法转化的植物中是有效的。尽管基于烟草摇铃病毒(TRV)的VIGS系统已经被开发并用于验证不同植物的基因功能，但还没有建立合适的TRV-VIGS方法来鉴定唐菖蒲。在这份报告中，我们描述了首次使用TRV-VIGS系统来沉默唐菖蒲的基因。利用GhPDS-VIGS载体对球茎和幼苗进行真空渗透处理，有效地下调了同源基因GhPDS的PHYTOENE去饱和酶基因的表达，并导致不同程度的光漂白。实时荧光定量RT-PCR检测TRV载体感染后GhPDS基因表达的降低。此外，利用pTRV2：CP-GFP载体对TRV感染进展进行了荧光可视化检测。综上所述，本文所描述的TRV介导的VIGS将是唐菖蒲基因功能分析的一种有效机制。
Functional analysis of genes in gladiolus has previously been impractical due to the lack of an efficient stable genetic transformation method. However, virus-induced gene silencing (VIGS) is effective in some plants which are difficult to transform through other methods. Although the Tobacco rattle virus (TRV)-based VIGS system has been developed and used for verifying gene functions in diverse plants, an appropriate TRV-VIGS approach for gladiolus has not been established yet. In this report we describe the first use of the TRV-VIGS system for gene silencing in gladiolus. Vacuum infiltration of cormels and young plants with the GhPDS-VIGS vector effectively down-regulated the PHYTOENE DESATURASE ortholog GhPDS gene and also resulted in various degrees of photobleaching in Gladiolus hybridus. The reduction in GhPDS expression was tested after TRV-based vector infection using real-time RT-PCR. In addition, the progress of TRV infection was detected by fluorescence visualization using a pTRV2: CP-GFP vector. In conclusion, the TRV-mediated VIGS described here will be an effective gene function analysis mechanism in gladiolus.