A woody plant degrading pathogen effector targets host glucose signaling protein VvRHIP1 to promote virulence

Lasiodiplodia theobromae is one of the causal agents of Grapevine trunk diseases, which becomes a tremendous threat to the grapevine production worldwide. Plant pathogens secrete diverse effectors to suppress host immune responses or regulate the host metabolism to promote diseases. Our results suggest that L. theobromae LtCre1 targets host VvRHIP1 to manipulate the sugar signaling pathway, through disrupting the association of VvRHIP1 and VvRGS1 complex. Overall design: Total RNA was isolated from N. benthamiana plant tissue using a TransZol Plant RNA Kit (TransGen Biotech, Beijing, China). The wild type seedings were used as control and samples grown under normal condition was collected for two biological replicates. The cDNA libraries were generated using NEBNext? Ultra? RNA Library Prep Kit for (Illumina, USA) according to manufacturer's recommendations. Then the libraries were sequenced using an Illumina Hiseq platform (Illumina, USA) and 150 bp paired-end reads were produced. Clean reads were filtered by removing reads containing ploy-N, reads containing adapter, and low-quality reads from the raw data. The filtered clean reads were aligned to the N. benthamiana reference genome using Hisat2 v2.0.4 with default parameters. The expression level for each transcript was normalized and estimated using the fragments per kilobases per million reads (FPKM) method (Trapnell et al., 2010). The differential expression analysis was determined using the DESeq R package (1.18.0) with default parameters (P-value <0.05; log2 (fold change) >2 or log2 (fold change) < -2). Gene Ontology (GO) terms for all differentially expressed genes was identified by the GOseq R package. The KEGG enrichment analysis of differential expression genes was performed using KOBAS software with default parameters.