Liu Shuai, Yu Yang, Guo Ke, Zhang Qian, Jia Zhaodong, Alfredo Morales Rodriguez, Ma Peiyong, Xie Hao, Bian Xiaofeng
Institute of Food Crops, Provincial Key Laboratory of Agrobiology, Jiangsu Academy of Agricultural Sciences, Nanjing, China.
Center for Tropical Crop Research, Research Institute of Tropical Roots and Tuber Crops (INIVIT), Santo Domingo, Cuba.
Front Plant Sci. 2024 Jul 16;15:1421244. doi: 10.3389/fpls.2024.1421244. eCollection 2024.
Griffithsin (GRFT) is a highly effective, broad-spectrum, safe, and stable viral inhibitor used to suppress a variety of viruses. However, little information is available on whether GRFT can prevent plant viral diseases. In this study, we constructed a GRFT overexpression vector containing the sweetpotato storage cell signal peptide and generated exogenous GRFT overexpression lines through genetic transformation. The transgenic plants showed notable resistance to sweetpotato virus disease in the virus nursery. To verify the antiplant virus function of GRFT, transient expression in tobacco leaves showed that GRFT inhibited the sweetpotato leaf curl virus (SPLCV). The replication of SPLCV was entirely inhibited when the concentration of GRFT reached a certain level. The results of pulldown and BIFC assays showed that GRFT did not interact with the six components of SPLCV. In addition, the mutated GRFT without the binding ability of carbohydrate and anticoronavirus function, in which three aspartate residues at carbohydrate binding sites were all mutated to alanine, also inhibited SPLCV. Quantitative reverse-transcription PCR analyses showed that the tobacco antiviral-related genes , , , and were overexpressed after GRFT/GRFT injection. Furthermore, , , and were more highly expressed in the leaves injected with GRFT. The results suggest that sweetpotato is able to express GRFT exogenously as a bioreactor. Moreover, exogenous GRFT expression inhibits plant viruses by promoting the expression of plant antiviral genes.
格里菲斯菌素(GRFT)是一种高效、广谱、安全且稳定的病毒抑制剂,用于抑制多种病毒。然而,关于GRFT是否能预防植物病毒病的信息却很少。在本研究中,我们构建了一个含有甘薯贮藏细胞信号肽的GRFT过表达载体,并通过遗传转化产生了外源GRFT过表达株系。转基因植株在病毒苗圃中对甘薯病毒病表现出显著抗性。为了验证GRFT的抗植物病毒功能,在烟草叶片中的瞬时表达表明GRFT抑制了甘薯卷叶病毒(SPLCV)。当GRFT浓度达到一定水平时,SPLCV的复制被完全抑制。下拉实验和双分子荧光互补实验结果表明,GRFT与SPLCV的六个组分均无相互作用。此外,在碳水化合物结合位点的三个天冬氨酸残基均突变为丙氨酸的、不具有碳水化合物结合能力和抗冠状病毒功能的突变型GRFT也能抑制SPLCV。定量逆转录PCR分析表明,注射GRFT/GRFT后,烟草抗病毒相关基因、、、和均过表达。此外,、和在注射GRFT的叶片中表达量更高。结果表明,甘薯能够作为生物反应器外源表达GRFT。此外,外源GRFT表达通过促进植物抗病毒基因的表达来抑制植物病毒。