National Center for Soybean Improvement, National Key Laboratory for Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Genetic Improvement of Soybean, Ministry of Agriculture, Nanjing Agricultural University, Weigang 1, Nanjing, China.
National Center for Soybean Improvement, National Key Laboratory for Crop Genetics and Germplasm Enhancement, Key Laboratory of Biology and Genetic Improvement of Soybean, Ministry of Agriculture, Nanjing Agricultural University, Weigang 1, Nanjing, China.
Virus Res. 2020 May;281:197870. doi: 10.1016/j.virusres.2020.197870. Epub 2020 Jan 18.
Soybean mosaic virus (SMV)-disease is one of the most serious and widespread diseases in soybean (Glycine max). In the present study, a DnaJ protein in soybean designated GmCPIP (SMV coat protein-interacting protein) was screened by the QIS-Seq (quantitative interactor screening with next-generation sequencing) method, and the interaction between SMV CP and GmCPIP was confirmed by the yeast two-hybrid (Y2H) system and bimolecular fluorescence complementation (BiFC) assay. Subcellular localization analysis indicated that both proteins are localized in the cytoplasm, cytomembrane and nucleus. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis showed that infection with SMV-SC4 temporarily increased the transcription of GmCPIP. Virus-induced gene silencing (VIGS) down-regulated the GmCPIP gene by 82%, and the accumulation of SMV was decreased by 88.6% in GmCPIP-silenced plants inoculated with SMV-SC4. The interaction of GmCPIP with SMV CP seems to contribute to SMV infection in soybean.
大豆花叶病毒(SMV)病是大豆(Glycine max)上最严重和最广泛的病害之一。在本研究中,采用 QIS-Seq(基于下一代测序的定量相互作用筛选)方法筛选了大豆中的一种 DnaJ 蛋白,命名为 GmCPIP(SMV 外壳蛋白相互作用蛋白),并通过酵母双杂交(Y2H)系统和双分子荧光互补(BiFC)实验证实了 SMV CP 与 GmCPIP 之间的相互作用。亚细胞定位分析表明,这两种蛋白均定位于细胞质、胞质膜和细胞核。实时定量聚合酶链反应(qRT-PCR)分析显示,SMV-SC4 感染会暂时增加 GmCPIP 的转录。通过病毒诱导的基因沉默(VIGS)下调 GmCPIP 基因 82%,并在接种 SMV-SC4 的 GmCPIP 沉默植株中,SMV 的积累减少了 88.6%。GmCPIP 与 SMV CP 的相互作用似乎有助于 SMV 在大豆中的感染。