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拟南芥抗西瓜花叶病毒基因rwm1的克隆揭示了天然病毒抗性基因的新功能。

Cloning of the Arabidopsis rwm1 gene for resistance to Watermelon mosaic virus points to a new function for natural virus resistance genes.

作者信息

Ouibrahim Laurence, Mazier Marianne, Estevan Joan, Pagny Gaëlle, Decroocq Véronique, Desbiez Cécile, Moretti André, Gallois Jean-Luc, Caranta Carole

机构信息

Genetics and Breeding of Fruits and Vegetables, INRA-UR1052, Dom. St Maurice, CS 60094, F-84143, Montfavet Cedex, France.

出版信息

Plant J. 2014 Sep;79(5):705-16. doi: 10.1111/tpj.12586. Epub 2014 Jul 25.

Abstract

Arabidopsis thaliana represents a valuable and efficient model to understand mechanisms underlying plant susceptibility to viral diseases. Here, we describe the identification and molecular cloning of a new gene responsible for recessive resistance to several isolates of Watermelon mosaic virus (WMV, genus Potyvirus) in the Arabidopsis Cvi-0 accession. rwm1 acts at an early stage of infection by impairing viral accumulation in initially infected leaf tissues. Map-based cloning delimited rwm1 on chromosome 1 in a 114-kb region containing 30 annotated genes. Positional and functional candidate gene analysis suggested that rwm1 encodes cPGK2 (At1g56190), an evolutionary conserved nucleus-encoded chloroplast phosphoglycerate kinase with a key role in cell metabolism. Comparative sequence analysis indicates that a single amino acid substitution (S78G) in the N-terminal domain of cPGK2 is involved in rwm1-mediated resistance. This mutation may have functional consequences because it targets a highly conserved residue, affects a putative phosphorylation site and occurs within a predicted nuclear localization signal. Transgenic complementation in Arabidopsis together with virus-induced gene silencing in Nicotiana benthamiana confirmed that cPGK2 corresponds to rwm1 and that the protein is required for efficient WMV infection. This work uncovers new insight into natural plant resistance mechanisms that may provide interesting opportunities for the genetic control of plant virus diseases.

摘要

拟南芥是理解植物对病毒病易感性潜在机制的一种有价值且高效的模式植物。在此,我们描述了在拟南芥Cvi-0生态型中负责对几种西瓜花叶病毒(WMV,马铃薯Y病毒属)分离株隐性抗性的一个新基因的鉴定和分子克隆。rwm1通过损害最初感染叶组织中的病毒积累在感染早期起作用。基于图谱的克隆将rwm1定位在1号染色体上一个114 kb的区域内,该区域包含30个注释基因。定位和功能候选基因分析表明,rwm1编码cPGK2(At1g56190),这是一种进化保守的核编码叶绿体磷酸甘油酸激酶,在细胞代谢中起关键作用。比较序列分析表明,cPGK2 N端结构域中的单个氨基酸替换(S78G)与rwm1介导的抗性有关。这种突变可能具有功能后果,因为它针对一个高度保守的残基,影响一个假定的磷酸化位点,并且发生在一个预测的核定位信号内。拟南芥中的转基因互补以及本氏烟草中的病毒诱导基因沉默证实,cPGK2对应于rwm1,并且该蛋白是WMV有效感染所必需的。这项工作揭示了对天然植物抗性机制的新见解,这可能为植物病毒病的遗传控制提供有趣的机会。

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