Establishment of a Virus-Induced Gene-Silencing (VIGS) System in Tea Plant and Its Use in the Functional Analysis of CsTCS1

• Published in: International journal of molecular sciences Vol. 24; no. 1; p. 392
• Main Authors: Li, Guodong, Li, Yan, Yao, Xinzhuan, Lu, Litang
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI 26-12-2022; MDPI AG
• Subjects: CsPOR1; CsTCS1; tea plant; tobacco rattle virus (TRV); virus-induced gene silencing (VIGS); CsPOR1; virus-induced gene silencing (VIGS); CsTCS1; tobacco rattle virus (TRV); tea plant
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms24010392

Tea (
[L.] O. Kuntze) is an important global economic crop and is considered to enhance health. However, the functions of many genes in tea plants are unknown. Virus-induced gene silencing (VIGS) mediated by tobacco rattle virus (TRV) is an effective tool for the analysis of gene functions, although this method has rarely been reported in tea plants. In this study, we established an effective VIGS-mediated gene knockout technology to understand the functional identification of large-scale genomic sequences in tea plants. The results showed that the VIGS system was verified by detecting the virus and using a real-time quantitative reverse transcription PCR (qRT-PCR) analysis. The reporter gene
(protochlorophyllide oxidoreductase) was silenced using the vacuum infiltration method, and typical photobleaching and albino symptoms were observed in newly sprouted leaves at the whole plant level of tea after infection for 12 d and 25 d. After optimization, the VIGS system was successfully used to silence the tea plant
(caffeine synthase) gene. The results showed that the relative caffeine content was reduced 6.26-fold compared with the control, and the level of expression of
decreased by approximately 3.12-fold in plants in which
was silenced. These results demonstrate that VIGS can be quickly and efficiently used to analyze the function of genes in tea plants. The successful establishment of VIGS could eliminate the need for tissue culture by providing an effective method to study gene function in tea plants and accelerate the process of functional genome research in tea.