Kim Nak Hyun, Lee Dong Hyuk, Choi Du Seok, Hwang Byung Kook
Department of Biology, The University of North Carolina at Chapel Hill, NC 27599-3280, USA.
Department of Plant Pathology, University of California Davis, Davis, CA 95616-8751, USA.
Plant Sci. 2015 Dec;241:307-15. doi: 10.1016/j.plantsci.2015.07.003. Epub 2015 Jul 29.
Carbohydrate-binding proteins, commonly referred to as lectins or agglutinins, function in defense responses to microbial pathogens. Pepper (Capsicum annuum) GNA-related lectin and PAN-domain protein gene CaGLP1 was isolated and functionally characterized from pepper leaves infected with Xanthomonas campestris pv. vesicatoria (Xcv). CaGLP1 contained an amine-terminus prokaryotic membrane lipoprotein lipid attachment site, a Galanthus nivalis agglutinin (GNA)-related lectin domain responsible for the recognition of high-mannose N-glycans, and a carboxyl-terminus PAN/apple domain. RNA gel blot and immunoblot analyses determined that CaGLP1 was strongly induced in pepper by compatible and incompatible Xcv infection. CaGLP1 protein localized primarily to the plasma membrane and exhibited mannose-binding specificity. CaGLP1-silenced pepper plants were more susceptible to compatible or incompatible Xcv infection compared with that of non-silenced control plants. CaGLP1 silencing in pepper leaves did not accumulate H2O2 and induce cell death during incompatible Xcv infection. Defense-related CaDEF1 (defensin) gene expression was significantly reduced in CaGLP1-silenced pepper plants. CaGLP1-overexpression in Arabidopsis thaliana enhanced resistance to Pseudomonas syringae pv. tomato. Defense-related AtPDF1.2 expression was elevated in CaGLP1-overexpression lines. Together, these results suggest that CaGLP1 is required for plant cell death and defense responses through the reactive oxygen species burst and downstream defense-related gene expression in response to bacterial pathogen challenge.
碳水化合物结合蛋白,通常被称为凝集素或凝集素,在对微生物病原体的防御反应中发挥作用。从感染野油菜黄单胞菌辣椒斑点病致病型(Xcv)的辣椒叶片中分离出辣椒(辣椒属)GNA相关凝集素和PAN结构域蛋白基因CaGLP1,并对其进行了功能表征。CaGLP1包含一个氨基末端原核细胞膜脂蛋白脂质附着位点、一个负责识别高甘露糖N聚糖的雪花莲凝集素(GNA)相关凝集素结构域,以及一个羧基末端PAN/苹果结构域。RNA凝胶印迹和免疫印迹分析确定,在辣椒中,CaGLP1在与Xcv的亲和性和非亲和性感染中均被强烈诱导。CaGLP1蛋白主要定位于质膜,并表现出甘露糖结合特异性。与未沉默的对照植株相比,CaGLP1沉默的辣椒植株对亲和性或非亲和性Xcv感染更敏感。在非亲和性Xcv感染期间,辣椒叶片中CaGLP1的沉默不会积累H2O2和诱导细胞死亡。在CaGLP1沉默的辣椒植株中,防御相关的CaDEF1(防御素)基因表达显著降低。在拟南芥中过表达CaGLP1可增强对丁香假单胞菌番茄致病型的抗性。在CaGLP1过表达株系中,防御相关的AtPDF1.2表达升高。总之,这些结果表明,CaGLP1通过活性氧爆发和下游防御相关基因表达参与植物细胞死亡和防御反应,以应对细菌病原体的挑战。