Genome-wide identification of glutamate receptor genes in adzuki bean and the roles of these genes in light and rust fungal response.

Plant glutamate receptor proteins (GLRs) are involved in plant development, biotic stress, and light-signal transduction. Vigna angularis is a traditional crop with important economic value in China, and the identification of functional genes can facilitate the breeding of stress resistant varieties. Here, we identified the members of the GLR gene family in the adzuki bean genome and investigated gene expression under light and rust fungal (Uromyces vignae) stimuli. Sixteen GLR genes were identified in V. angularis (VaGLRs), and these genes clustered in a single clade (clade III) with two groups. Evolutionary analysis showed that three VaGLRs result from tandem duplications and four result from whole genome/segmental duplications. To understand the regulation of expression of VaGLRs, cis-acting elements were analyzed in the promoter regions of the VaGLRs including cis-acting elements associated with light and stress responsiveness. Expression analysis by qRT-PCR revealed transcripts of eight and 10 VaGLRs in response to light stimuli and rust infection, respectively. For light responsiveness, the expression levels of XP_017430569.1 and XP_017425299.1 were higher under light condition than in darkness, while the expression levels of XP_017406996.1, XP_017425763.1, and XP_017423557.1 gradually recovered during dark treatment. Additionally, the relative expression levels of XP_017413816.1, XP_017436268.1, and XP_017425299.1 were significantly elevated during U. vignae infection in a resistant cultivar compared to the expression levels in a susceptible cultivar. XP_017425299.1 expression was induced both by light and rust infection, suggesting this gene may link light and disease resistance signaling pathways. Our results provide insight into how the VaGLRs contribute to adzuki bean response to light stimulus and pathogen attack. These identified VaGLRs also provide important reference to improve adzuki bean germplasm resources.

A natriuretic peptide molecule from Vigna angularis, VaEG45, confers rust resistance by inhibiting fungal development.

KEY MESSAGE: Novel function and mechanism of a PNP molecule VaEG45 from adzuki bean involved in plant immunity. Plant natriuretic peptides (PNPs) can affect a broad spectrum of physiological responses in plants acting as peptidic signaling molecules. However, PNPs may play additional roles in plant immunity. Our previous transcriptome data of adzuki bean (Vigna angularis) in response to Uromyces vignae infection revealed association of PNP-encoding gene VaEG45 with U. vignae resistance. To determine the function of VaEG45 in disease resistance, we cloned the 589 bp nucleotide sequence of VaEG45 containing 2 introns, encoding a putative 13.68 kDa protein that is 131 amino acids in length. We analyzed expression in different resistant cultivars of V. angularis and found significant induction of VaEG45 expression after U. vignae infection. Transient expression of VaEG45 improved tobacco resistance against Botrytis cinerea. We next analyzed the mechanism by which VaEG45 protects plants from fungal infection by determination of the biological activity of the prokaryotic expressed VaEG45. The results showed that the fusion protein VaEG45 can significantly inhibit urediospores germination of U. vignae, mycelial growth, and the infection of tobacco by B. cinerea. Further analysis revealed that VaEG45 exhibits ß-1, 3-glucanase activity. These findings uncover the function of a novel PNP molecule VaEG45 and provide new evidence about the mechanism of PNPs in plant immunity.

Histological and molecular responses of Vigna angularis to Uromyces vignae infection.

BACKGROUND: To advance the understanding of adzuki bean (Vigna angularis) resistance to infection with the rust-causing fungus Uromyces vignae (Uv), we comprehensively analyzed histological events and the transcriptome of Uv-infected adzuki bean. RESULTS: Compared with the susceptible cv. Baoqinghong (BQH), the resistant cv. QH1 showed inhibition of uredospore germination and substomatal vesicle development, intense autofluorescence of cells around the infection site, and cell wall deposit formation in response to Uv infection. In cv. QH1, gene set enrichment analysis (GSEA) showed enrichment of chitin catabolic processes and responses to biotic stimuli at 24 h post-inoculation (hpi) and cell wall modification and structural constituent of cytoskeleton at 48 hpi. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated enrichment of WRKY transcription factors (TFs), the calcium binding protein cml, and hydroquinone glucosyltransferase at both 24 and 48 hpi. In total, 1992 and 557 differentially expressed genes (DEGs) were identified at 24 and 48 hpi, respectively. Cell surface pattern-recognition receptors (PRRs), WRKY TFs, defense-associated pathogenesis-related (PR) proteins, and lignin and antimicrobial phenolic compound biosynthesis were significantly induced. Finally, we detected the chitinase (CHI) and phenylalanine ammonia-lyase (PAL) activity were higher in QH1 and increased much earlier than in BQH. CONCLUSION: In cv. QH1, cell-surface PRRs rapidly recognize Uv invasion and activate the corresponding TFs to increase the transcription of defense-related genes and corresponding enzymatic activities to prevent fungal development and spread in host tissues.