High Level Expression of a Virus Resistance Gene, RCY1, Confers Extreme Resistance to Cucumber mosaic virus in Arabidopsis thaliana

Abstract

A coiled coil-nucleotide binding site-leucine rich repeat-type resistance gene, RCY1, confers resistance to a yellow strain of Cucumber mosaic virus, CMV(Y), in Arabidopsis thaliana ecotype C24. Resistance to CMV(Y) in C24 is accompanied by a hypersensitive response (HR) that is characterized by the development of necrotic local lesions at the primary infection sites. To further study the HR and resistance to CMV(Y) in ecotype Col-0, which is susceptible to CMV(Y), Col-0 were transformed with RCY1. Systemic spread of CMV(Y) was completely suppressed in RCY1-transformed Col-0 (Col::pRCY1 lines 2 to 6), whereas virulent strain CMV(B2) spread and multiplied systemically in these transgenic lines similar to that in wild-type Col-0. Interestingly, the resistant phenotype of Col::pRCY1 varied among the lines. In lines 3 and 6, in which levels of RCY1 transcript were similar to that in wild-type C24, the HR and resistance to CMV(Y) was induced. Line 4, which expresses moderately elevated levels of RCY1 transcript, exhibited moderately enhanced resistance compared with that in C24 or line 3. In contrast, lines 2 and 5, which highly overexpress the RCY1 gene, did not exhibit either visible lesions or a micro-HR on the inoculated leaves. Moreover, virus coat protein was not detected in either inoculated or noninoculated upper leaves of these two lines, suggesting that extreme resistance (ER) to CMV(Y) was induced by high levels of expression of RCY1. Furthermore, in transgenic lines expressing hemagglutinin (HA) epitope-tagged RCY1 (Col::pRCY1-HA), high levels of accumulation of RCY1-HA protein were also correlated with the ER phenotype. Global gene expression analysis in line 2, which highly overexpresses RCY1, indicated that expression of several defense-related genes were constitutively elevated compared with wild-type Col-0. Despite this, line 2 did not have enhanced resistance to other avirulent and virulent pathogens. Take together, constitutive accumulation of high levels of RCY1 protein appears to regulate the strength of RCY1-conferred resistance in a gene-for-gene manner and implies that ER and HR-associated resistance differ only in the strength of resistance.