Integrated Transcriptomic and Metabolomic Analysis Reveals GHABF3-Mediated ABA Signaling Governs Enhanced...

Verticillium wilt (VW) is a soil-borne fungal plant disease. Gossypium hirsutum varieties with the widest planting area are highly susceptible to VW pathogens, because their narrow genetic background of germplasm resources causes difficulties in cultivating VW-resistant varieties through intraspecific breeding. Therefore, G. barbadense cultivars, harboring a natural VW resistance, become ideal donor materials to cultivate high-yield and multi-resistance chromosome segment substitution lines (CSSLs) through hybridization and backcrossing with G. hirsutum receptor and recurrent parent. In order to investigate the molecular mechanism of cotton response to VW infection, a BC₅F_3:5 CSSL MBI9626 and its parents, CCRI36 (G. hirsutum) and Hai1 (G. barbadense), were chosen to perform transcriptome and metabolome sequencing on their root samples at 0, 7, and 15 days after inoculation (DAI) of V. dahliae V991. In total, 36,564 differentially expressed genes (DEGs) and 102 differentially accumulated metabolites (DAMs) were separately identified from 12 pairwise comparison groups among the 27 samples. Of those, 125 common DEGs were found to participate in the biological processes of oxylipin metabolism, jasmonic acid (JA) biosynthesis/metabolism, and response to wounding in Gene Ontology (GO) enrichment analyses, while most of the DAMs were significantly enriched in tyrosine, purine, and phenylalanine metabolism pathways in enrichment analyses of Kyoto Encyclopedia of Genes and Genomes (KEGG). Having performed a conjoint KEGG analysis of all the DEGs and DAMs, we found two commonly enriched pathways, namely plant hormone signal transduction and flavonoid biosynthesis, which were consistent with the enrichment annotations of the significant model in weighted gene co-expression network analysis on the 2091 DEGs identified by an intersection of the genes in 40 previous QTLs and the total DEGs of this RNA-seq data. Among the ABA signaling pathway, the gene GH_D12G0236 (GHABF3) was selected to be used to perform virus-induced gene silencing (VIGS) verification in CCRI36 and MBI9626, and GHABF3-silenced plants showed a more serious wilting phenotype, an increased disease index (DI), and higher accumulation of fungal biomass compared to their empty-vector plants. These results provide a high-efficiency strategy for screening vital genes affecting cotton VW resistance, and lay a solid foundation for further cotton molecular breeding.