Chinese Wheat Mosaic Virus-Induced Gene Silencing in Monocots and Dicots at Low Temperature

文献类型： 外文期刊

第一作者： Yang, Jian

作者： Yang, Jian;Zhang, Tian-Ye;He, Long;Li, Juang;Chen, Xuan;Chen, Jian-Ping;Liao, Qian-Sheng;Zhang, Heng-Mu;Li, Jing;Yang, Jin;Zhang, Heng-Mu;Li, Jing;Yang, Jin;Li, Jin-Bang

作者机构：

关键词： RNA virus; gene silecning; Chinese wheat mosaic virus; vector; low temperature

期刊名称：FRONTIERS IN PLANT SCIENCE （ 影响因子：5.753； 五年影响因子：6.612 ）

ISSN： 1664-462X

年卷期： 2018 年 9 卷

页码：

收录情况： SCI

摘要： Virus-induced gene silencing (VIGS) is an important tool for functional genomics studies in plants. With this method, it is possible to target most endogenous genes and downregulate the messenger RNA (mRNA) in a sequence-specific manner. Chinese wheat mosaic virus (CWMV) has a bipartite, single-strand positive RNA genome, and can infect both wheat and Nicotiana benthamiana, and the optimal temperature for systemic infection in plants is 17 degrees C. To assess the potential of the virus as a vector for gene silencing at low temperature, a fragment of the N. benthamiana or wheat phytoene desaturase (PDS) gene was expressed from a modified CWMV RNA2 clone and the resulting photo bleaching in infected plants was used as a reporter for silencing. Downregulation of PDS mRNA was also measured by quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR). In experiments using fragments of PDS ranging from 500 to 1500 nucleotides, insert length influenced the stability and the efficiency of VIGS. The CWMV induced silencing system was also used to suppress miR165/166 and miR3134a through expression of miRNA target mimics. The relative expression levels of mature miR165/166 and miR3134a decreased whereas the transcript levels of their target genes increased. Interestingly, we also found the CWMV-induced silencing system was more efficient compare with the vector based on Barley stripe mosaic virus (BSMV) or Foxtail mosaic virus (FoMV) in wheat or the vector based on TRV in N. benthamiana at 17 degrees C. In summary, the CWMV vector is effective in silencing endogenous genes and miRNAs at 17 degrees C, thereby providing a powerful tool for gene function analysis in both N. benthamiana and wheat at low temperature.

分类号：

  • 相关文献
作者其他论文 更多>>

微信小程序