Tobacco rattle virus-inducedPHYTOENE DESATURASE(PDS)andMg-chelatase H subunit(ChlH) gene silencing inSolanum pseudocapsicumL.

Abstract

Virus-induced gene silencing (VIGS) is an attractive tool for determining gene function in plants. The present study constitutes the first application of VIGS in S. pseudocapsicum, which has great ornamental and pharmaceutical value, using tobacco rattle virus (TRV) vectors. Two marker genes, PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH), were used to test the VIGS system in S. pseudocapsicum. The photobleaching and yellow-leaf phenotypes of the silenced plants were shown to significantly correlate with the down-regulation of endogenous SpPDS and SpChlH, respectively (P ≤ 0.05). Moreover, the parameters potentially affecting the efficiency of VIGS in S. pseudocapsicum, including the Agrobacterium strain and the inoculation method (leaf syringe-infiltration, sprout vacuum-infiltration and seed vacuum-infiltration), were compared. The optimized VIGS parameters were the leaf syringe-infiltration method, the Agrobacterium strain GV3101 and the growth of agro-inoculated plants at 25°. With these parameters, the silencing efficiency of SpPDS and SpChlH could reach approximately 50% in S. pseudocapsicum. Additionally, the suitability of various reference genes was screened by RT-qPCR using three candidate genes, and the results demonstrated that glyceraldehyde 3-phosphate dehydrogenase (GAPDH) can serve as a suitable reference for assessing the gene expression levels of VIGS systems in S. pseudocapsicum. The proven application of VIGS in S. pseudocapsicum and the characterization of a suitable reference gene in the present work will expedite the functional characterization of novel genes in S. pseudocapsicum.

Virus-induced gene silencing (VIGS) in cape gooseberry (Physalis peruviana L., Solanaceae)

“…From the 20 inoculated plants with the pTRV2:PpPDS vector, 10 plants showed photo-bleaching leaves indicating a rate efficiency of 50 %, which is consistent with other reports [40,42,43] (Fig. 4).…”

An efficient virus-induced gene silencing (VIGS) system for functional genomics in Brassicas using a cabbage leaf curl virus (CaLCuV)-based vector

“…According to our results, it did not affect our judgment to silencing phenotype. We found that described above silencing spots also appeared in other VIGS experiments, S. pseudocapsicum, cotton and pepper sliencing by TRV, N. benthamiana sliencing by CaLCuV, and so on [22,[35][36]65].…”

“…Tuttle et al discovered that the ambient temperature needed to be lower than 21 °C to obtain a better silencing phenotype and that TRV-mediated VIGS was almost completely inhibited when the ambient temperature was higher than 28 °C [35]. Xu et al (2018) tested silencing efficiency by TRV under different growth temperatures (18 °C, 25 °C and 30 °C), and the speed of the response and the efficiency of infection were optimal at 25 °C [36]. In our study, the 22 °C is the most suitable temperature for PCVA/PVCB in cabbage, suggesting that appropriate low temperature can improve silence efficiency.…”

“…Moreover, the silencing efficiency of VIGS is influenced by many parameters, including vector targeting fragment (insertion direction and the size of the insert sequence) [18,32], infection pattern [33][34], culture environment [35][36] and plant growth stage [37]. Therefore, more research and exploration are needed for more efficient VIGS systems.…”

Host-Induced Gene Silencing of an Adenylate Kinase Gene Involved in Fungal Energy Metabolism Improves Plant Resistance to Verticillium dahliae

“…Recently, HIGS has also proved to be a promising strategy for silencing a foreign gene in a host to investigate numerous plant-pathogen systems [61,62], including Vd in various hosts [33,34,63]. In the present study, positive plants at 10 days after infiltration showed distinct photobleaching, suggesting the massive presence of dsRNA and its active role in gene silencing [64][65][66]. In our study, seedlings that were inoculated with Vd developed varying degrees of wilting from 10 to 12 dpi.…”