A single-nucleotide mutation in GLUTAMATE RECEPTOR-LIKE protein gene confers resistance to Fusarium wilt in Gossypium hirsutum

Fusarium wilt (FW) disease of cotton, caused by the fungus Fusarium oxysporum f. sp. vasinfectum (Fov), causes severe losses in cotton production worldwide. Though significant advancements have been made in development of FW‐resistant Upland cotton (Gossypium hirsutum) in resistance screening programs, the precise resistance genes and the corresponding molecular mechanisms for resistance to Fov remain unclear. Herein it is reported that Fov7, a gene unlike canonical plant disease‐resistance (R) genes, putatively encoding a GLUTAMATE RECEPTOR‐LIKE (GLR) protein, confers resistance to Fov race 7 in Upland cotton. A single nucleotide polymorphism (SNP) (C/A) in GhGLR4.8, resulting in an amino acid change (L/I), is associated with Fov resistance. A PCR‐based DNA marker (GhGLR4.8SNP(A/C)) is developed and shown to cosegregate with the Fov resistance. CRISPR/Cas9‐mediated knockout of Fov7 results in cotton lines extremely susceptible to Fov race 7 with a loss of the ability to induce calcium influx in response to total secreted proteins (SEPs) of Fov. Furthermore, coinfiltration of SEPs with GhGLR4.8A results in a hypersensitive response. This first report of a GLR‐encoding gene that functions as an R gene provides a new insight into plant–pathogen interactions and a new handle to develop cotton cultivars with resistance to Fov race 7

Verticillium alfalfae and V. dahliae, Agents of Verticillium Wilt Diseases

Verticillium dahliae and V. alfalfae (formerly Verticillium albo-atrum) are two important agricultural pathogens that affect many crops around the world and cause a distinct type of vascular wilt, which are known as Verticillium wilts. Several V. alfalfae and V. dahliae genomes have been sequenced, and are among the smaller genomes from filamentous ascomycetes. The number of predicted protein-encoding genes is similar to the saprobe Neurospora crassa. Perhaps reflective of their particular hemibiotrophic life styles, some gene families are expanded in the V. alfalfae and V. dahliae genomes. These include the gene families encoding glycoside hydrolases GH88, necrosis and ethylene-inducing-like proteins (NLPs), LysM effectors, proteins with chitin-recognition motifs, and cutinases. But the number of predicted secreted proteins was less than half that of the related Colletotrichum species, the agents of anthracnose diseases. V. dahliae strains generally contain lineage-specific regions (LS regions), which may play an important role in virulence and pathogenicity. Examples for horizontal transfer into Verticillium ancestors include the virulence factor Ave1, a glucan glucosyltransferase, and potentially some of the retrotransposons. The V. alfalfae and V. dahliae genomes have already had significant impacts on various aspects of basic and applied Verticillium research

GhWRKY41 forms a positive feedback regulation loop and increases cotton defence response against Verticillium dahliae by regulating phenylpropanoid metabolism

Despite the established significance of WRKY proteins and phenylpropanoid metabolism in plant immunity, how WRKY proteins modulate aspects of the phenylpropanoid pathway remains undetermined. To understand better the role of WRKY proteins in plant defence, we identified a cotton (Gossypium hirsutum) protein, GhWRKY41, that is, universally and rapidly induced in three disease-resistant cotton cultivars following inoculation with the plant pathogenic fungus, Verticillium dahliae. We show that overexpression of GhWRKY41 in transgenic cotton and Arabidopsis enhances resistance to V. dahliae, while knock-down increases cotton more susceptibility to the fungus. GhWRKY41 physically interacts with itself and directly activates its own transcription. A genome-wide chromatin immunoprecipitation and high-throughput sequencing (ChIP-seq), in combination with RNA sequencing (RNA-seq) analyses, revealed that 43.1% of GhWRKY41-binding genes were up-regulated in cotton upon inoculation with V. dahliae, including several phenylpropanoid metabolism master switches, receptor kinases, and disease resistance-related proteins. We also show that GhWRKY41 homodimer directly activates the expression of GhC4H and Gh4CL, thereby modulating the accumulation of lignin and flavonoids. This finding expands our understanding of WRKY-WRKY protein interactions and provides important insights into the regulation of the phenylpropanoid pathway in plant immune responses by a WRKY protein

Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis.

Ustilago maydis is a ubiquitous pathogen of maize and a well-established model organism for the study of plant-microbe interactions. This basidiomycete fungus does not use aggressive virulence strategies to kill its host. U. maydis belongs to the group of biotrophic parasites (the smuts) that depend on living tissue for proliferation and development. Here we report the genome sequence for a member of this economically important group of biotrophic fungi. The 20.5-million-base U. maydis genome assembly contains 6,902 predicted protein-encoding genes and lacks pathogenicity signatures found in the genomes of aggressive pathogenic fungi, for example a battery of cell-wall-degrading enzymes. However, we detected unexpected genomic features responsible for the pathogenicity of this organism. Specifically, we found 12 clusters of genes encoding small secreted proteins with unknown function. A significant fraction of these genes exists in small gene families. Expression analysis showed that most of the genes contained in these clusters are regulated together and induced in infected tissue. Deletion of individual clusters altered the virulence of U. maydis in five cases, ranging from a complete lack of symptoms to hypervirulence. Despite years of research into the mechanism of pathogenicity in U. maydis, no 'true' virulence factors had been previously identified. Thus, the discovery of the secreted protein gene clusters and the functional demonstration of their decisive role in the infection process illuminate previously unknown mechanisms of pathogenicity operating in biotrophic fungi. Genomic analysis is, similarly, likely to open up new avenues for the discovery of virulence determinants in other pathogens. ©2006 Nature Publishing Group