Zhang Yao, Niu Nazi, Li Shijia, Liu Yin, Xue Chaoling, Wang Huibin, Liu Mengjun, Zhao Jin
College of Life Science, Hebei Agricultural University, Baoding 071000, China.
Research Center of Chinese Jujube, Hebei Agricultural University, Baoding 071000, China.
Plants (Basel). 2023 May 26;12(11):2115. doi: 10.3390/plants12112115.
Virus-induced gene silencing (VIGS) is a fast and efficient method for assaying gene function in plants. At present, the VIGS system mediated by Tobacco rattle virus (TRV) has been successfully practiced in some species such as cotton and tomato. However, little research of VIGS systems has been reported in woody plants, nor in Chinese jujube. In this study, the TRV-VIGS system of jujube was firstly investigated. The jujube seedlings were grown in a greenhouse with a 16 h light/8 h dark cycle at 23 °C. After the cotyledon was fully unfolded, mixture containing pTRV1 and pTRV2-ZjCLA with OD = 1.5 was injected into cotyledon. After 15 days, the new leaves of jujube seedlings showed obvious photo-bleaching symptoms and significantly decreased expression of , indicating that the TRV-VIGS system had successfully functioned on jujube. Moreover, it found that two injections on jujube cotyledon could induce higher silencing efficiency than once injection. A similar silencing effect was then also verified in another gene, . These results indicate that the TRV-VIGS system in Chinese jujube has been successfully established and can be applied to evaluate gene function, providing a breakthrough in gene function verification methods.
病毒诱导的基因沉默(VIGS)是一种在植物中快速高效地检测基因功能的方法。目前,由烟草脆裂病毒(TRV)介导的VIGS系统已在棉花和番茄等一些物种中成功应用。然而,关于VIGS系统在木本植物中,尤其是在枣树中的研究报道较少。在本研究中,首次对枣树的TRV-VIGS系统进行了研究。枣树苗在温室中生长,温度为23°C,光照/黑暗周期为16小时/8小时。子叶完全展开后,将OD = 1.5的含有pTRV1和pTRV2-ZjCLA的混合物注射到子叶中。15天后,枣树苗的新叶出现明显的光漂白症状,且 的表达显著降低,表明TRV-VIGS系统在枣树上成功发挥了作用。此外,还发现对枣树子叶进行两次注射比一次注射能诱导更高的沉默效率。随后在另一个基因 中也验证了类似的沉默效果。这些结果表明,枣树的TRV-VIGS系统已成功建立,可用于评估基因功能,为基因功能验证方法提供了突破。