caa a22 4500 467895406 CHVBK 20180406152802.0 cr unu---uuuuu 170328e20060901xx s 000 0 eng 10.1007/s11103-006-9016-z doi (NATIONALLICENCE)springer-10.1007/s11103-006-9016-z Development of a Virus-Induced Gene-Silencing System for Functional Analysis of the RPS2 -Dependent Resistance Signalling Pathways in Arabidopsis [Elektronische Daten] [Xin-Zhong Cai, Qiu-Fang Xu, Chang-Chun Wang, Zhong Zheng] Virus-induced gene silencing (VIGS) offers a rapid and high throughput technique platform for the analysis of gene function in plants. Although routinely used in some Solanaceous species, VIGS system has not been well established in Arabidopsis thaliana (L.) Heynh. We have recently reported some factors that potentially influence tobacco rattle virus (TRV)-mediated VIGS of phytoene desaturase (PDS) and actin gene expression in Arabidopsis. In this study, we have further established that the Agrobacterium strain used for agro-inoculation significantly affects the VIGS efficiency. Strain GV3101 was highly effective; C58C1 and LBA4404 were invalid, while EHA105 was plant growth stage-dependent for TRV-induced gene silencing. Furthermore, the VIGS procedure optimised for the PDS gene was applied for the functional analysis of the disease resistance gene RPS2-mediated resistance pathway. Silencing of RPS2 led to loss of resistance to the otherwise avirulence strain of Pseudomonas syringae pv. tomato DC3000 carrying the avirulence gene AvrRpt2. Silencing of RIN4, a RPS2 repressor gene, gave rise to conversion of compatible interaction to incompatible. Silencing of NDR1, RAR1 and HSP90, known to be required for the RPS2-mediated resistance, resulted in loss of the resistance, while silencing of EDS1 and SGT1b, which are not required for the RPS2-mediated resistance, caused no change of the resistance. These results indicate that the optimised procedure for the TRV-based VIGS is a potentially powerful tool for dissecting the signal transduction pathways of disease resistance in Arabidopsis. Springer Science+Business Media B.V., 2006 Arabidopsis thaliana nationallicence Disease resistance nationallicence RPS2 nationallicence Signal transduction nationallicence Tobacco rattle virus (TRV) nationallicence Virus-induced gene silencing (VIGS) nationallicence Agro-inoculation : Agrobacterium inoculation nationallicence cfu : colony-forming units nationallicence dpa : days post agro-inoculation nationallicence dpi : days post inoculation nationallicence EST : expressed sequence tag nationallicence OD : optical density nationallicence PDS : phytoene desaturase gene nationallicence Pst : Pseudomonas syringae pv. tomato nationallicence TRV : tobacco rattle virus nationallicence VIGS : virus-induced gene silencing nationallicence Cai Xin-Zhong Institute of Biotechnology, and Department of Plant Protection, Zhejiang University, 268 Kai Xuan Road, 310029, Hangzhou, P.R. China aut Xu Qiu-Fang Institute of Biotechnology, and Department of Plant Protection, Zhejiang University, 268 Kai Xuan Road, 310029, Hangzhou, P.R. China aut Wang Chang-Chun Institute of Biotechnology, and Department of Plant Protection, Zhejiang University, 268 Kai Xuan Road, 310029, Hangzhou, P.R. China aut Zheng Zhong Institute of Biotechnology, and Department of Plant Protection, Zhejiang University, 268 Kai Xuan Road, 310029, Hangzhou, P.R. China aut Plant Molecular Biology Kluwer Academic Publishers 62/1-2(2006-09-01), 223-232 0167-4412 62:1-2<223 2006 62 11103 https://doi.org/10.1007/s11103-006-9016-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11103-006-9016-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Cai Xin-Zhong Institute of Biotechnology, and Department of Plant Protection, Zhejiang University, 268 Kai Xuan Road, 310029, Hangzhou, P.R. China aut NATIONALLICENCE 700 1- Xu Qiu-Fang Institute of Biotechnology, and Department of Plant Protection, Zhejiang University, 268 Kai Xuan Road, 310029, Hangzhou, P.R. China aut NATIONALLICENCE 700 1- Wang Chang-Chun Institute of Biotechnology, and Department of Plant Protection, Zhejiang University, 268 Kai Xuan Road, 310029, Hangzhou, P.R. China aut NATIONALLICENCE 700 1- Zheng Zhong Institute of Biotechnology, and Department of Plant Protection, Zhejiang University, 268 Kai Xuan Road, 310029, Hangzhou, P.R. China aut NATIONALLICENCE 773 0- Plant Molecular Biology Kluwer Academic Publishers 62/1-2(2006-09-01), 223-232 0167-4412 62:1-2<223 2006 62 11103 Metadata rights reserved Springer special CC-BY-NC licence nationallicence SWISSBIB 462662934 BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer