Abstract

Ubiquitination is essential for ubiquitin/proteasome-mediated protein degradation in plant development and defense. Here, we identified a novel E3 ubiquitin ligase RING1 gene, CaRING1, from pepper (Capsicum annuum). In pepper, CaRING1 expression is induced by avirulent Xanthomonas campestris pv vesicatoria infection. CaRING1 contains an amino-terminal transmembrane domain and a carboxyl-terminal RING domain. In addition, it displays in vitro E3 ubiquitin ligase activity, and the RING domain is essential for E3 ubiquitin ligase activity in CaRING1. CaRING1 also localizes to the plasma membrane. In pepper plants, virus-induced gene silencing of CaRING1 confers enhanced susceptibility to avirulent X. campestris pv vesicatoria infection, which is accompanied by compromised hypersensitive cell death, reduced expression of PATHOGENESIS-RELATED1, and lowered salicylic acid levels in leaves. Transient expression of CaRING1 in pepper leaves induces cell death and the defense response that requires the E3 ubiquitin ligase activity of CaRING1. By contrast, overexpression of CaRING1 in Arabidopsis (Arabidopsis thaliana) confers enhanced resistance to hemibiotrophic Pseudomonas syringae pv tomato and biotrophic Hyaloperonospora arabidopsidis infections. Taken together, these results suggest that CaRING1 is involved in the induction of cell death and the regulation of ubiquitination during the defense response to microbial pathogens.

Highlights

  • Ubiquitination is essential for ubiquitin/proteasome-mediated protein degradation in plant development and defense

  • The RING domain belongs to the conserved C3H2C3-type RING-H2 group, which are essential for E3 ubiquitin ligase activity in the ubiquitin/26S proteasome system

  • We have found that the pepper E3 ubiquitin ligase RING1 (CaRING1) gene is required for disease resistance and for the hypersensitive response (HR) cell death against Xanthomonas campestris pv vesicatoria (Xcv) infection

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Summary

Introduction

Ubiquitination is essential for ubiquitin/proteasome-mediated protein degradation in plant development and defense. Overexpression of CaRING1 in Arabidopsis (Arabidopsis thaliana) confers enhanced resistance to hemibiotrophic Pseudomonas syringae pv tomato and biotrophic Hyaloperonospora arabidopsidis infections Taken together, these results suggest that CaRING1 is involved in the induction of cell death and the regulation of ubiquitination during the defense response to microbial pathogens. In Arabidopsis cell cultures and seedlings, the general bacterial flagellin peptide elicitor (flg22), which stimulates basal defense responses, can induce a 2.5-fold change in the expression levels of over 250 genes (Felix et al, 1999) Among these up-regulated genes were genes encoding 10 putative RING finger E3 ligases, including RING-H2 FINGER A3b (RHA3b), RHA1b, RING MEMBRANE-ANCHOR1, and ATL6 (Navarro et al, 2004). The Arabidopsis E3 ubiquitin ligases PLANT U-BOX PROTEIN22 (PUB22), PUB23, and PUB24 play roles in the negative regulation of pathogen-associated molecular patterns-triggered immunity (Trujillo et al, 2008)

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