First, second, and fourth authors: School of Life Science and Biotechnology, Dalian University of Technology, Dalian, 116024, China; and third author: School of Computer Science and Technology, Dalian University of Technology, Dalian 116024, China.
Phytopathology. 2018 Aug;108(8):980-987. doi: 10.1094/PHYTO-12-17-0389-R. Epub 2018 Jun 14.
The nucleotide binding sites-leucine-rich repeat (NBS-LRR) genes are key regulatory components of plant to pathogens. Phytophthora infestans-inducible coding sequence encoding an NBS-LRR (SpNBS-LRR) protein in tomato (Solanum pimpinellifolium L3708) was cloned and characterized based on our RNA-Seq data and tomato genome. After sequence analysis, SpNBS-LRR was identified as a hydrophilic protein with no transmembrane topological structure and no signal peptide. SpNBS-LRR had a close genetic relationship to RPS2 of Arabidopsis thaliana by phylogenetic analysis. In addition, SpNBS-LRR gene was mainly expressed in root, with low expression observed in leaf and stem. To further investigate the role of SpNBS-LRR in tomato-P. infestans interaction, SpNBS-LRR was introduced in susceptible tomatoes and three transgenic lines with higher expression level of SpNBS-LRR were selected. These transgenic tomato plants that overexpressed SpNBS-LRR displayed greater resistance than wild-type tomato plants after infection with P. infestans, as shown by decreased disease index, lesion diameters, number of necrotic cells, P. infestans abundance, and higher expression levels of the defense-related genes. This information provides insight into SpNBS-LRR involved in the resistance of tomato to P. infestans infection and candidate for breeding to enhance biotic stress-resistance in tomato.
核苷酸结合位点-富含亮氨酸重复(NBS-LRR)基因是植物与病原体相互作用的关键调节因子。基于我们的 RNA-Seq 数据和番茄基因组,克隆并鉴定了番茄(Solanum pimpinellifolium L3708)中与疫霉菌诱导相关的 NBS-LRR(SpNBS-LRR)编码序列。序列分析表明,SpNBS-LRR 是一种亲水蛋白,无跨膜拓扑结构,无信号肽。系统进化分析表明,SpNBS-LRR 与拟南芥 RPS2 具有密切的遗传关系。此外,SpNBS-LRR 基因在根中主要表达,在叶和茎中表达水平较低。为了进一步研究 SpNBS-LRR 在番茄与疫霉菌互作中的作用,将 SpNBS-LRR 导入感病番茄中,并选择 SpNBS-LRR 表达水平较高的三个转基因株系。与野生型番茄植株相比,过表达 SpNBS-LRR 的转基因番茄植株在感染疫霉菌后表现出更强的抗性,表现为病情指数降低、病斑直径减小、坏死细胞数量减少、疫霉菌丰度降低以及防御相关基因表达水平升高。这些结果为 SpNBS-LRR 参与番茄对疫霉菌侵染的抗性提供了新的认识,并为培育提高番茄生物胁迫抗性的品种提供了候选基因。