Analysis of structure and differential expression of Pseudomonas syringae 5-like (RPS5-like) genes in pathogen-infected Vitis flexuosa

摘要

Nucleotide binding site-leucine-rich repeat (NBS-LRR) genes are the largest gene resistance family in plants. Resistance to Pseudomonas syringae 5 (RPS5) is a member of the NBS-LRR family. RPS5 and its homologs are important disease resistance genes in plants including Arabidopsis. This study identified nine loci of RPS5-like genes in Vitis flexuosa that showed differential expression from transcriptome analysis by next generation sequencing (NGS) of V. flexuosa infected with Elsinoe ampelina based on structural analyses: VfRPS5-like1059 (Vitis flexuosa resistance to Pseudomonas syringae 5-like 1059), VfRPS5-like1833, VfRPS5-like4135, VfRPS5-like4833, VfRPS5-like6172, VfRPS5-like13564, VfRPS5-like20585, VfRPS5-like55532, and VfRPS5-like62178. These genes, 2236-4762 bp in length, exhibited the structure of a coiled coil-nucleotide binding site-leucine-rich repeat (CC-NBS-LRR) resistance gene. The predicted amino acid sequences of all nine genes contained typical NBS domains. Chromosomal localization revealed that the nine genes were located in six chromosomes of the V. flexuosa pseudo-genome. Of the nine loci, only VfRPS5-like55532 showed upregulated expression against all tested pathogens at all tested time points, except 24 h postinoculation (hpi) for E. ampelina. All tested genes also showed differential expression against major grapevine fungal and bacterial pathogens. The results presented herein will also serve as a useful reference dataset for functional analysis and utilization in molecular breeding of resistant grapevines.