The Ustilago maydis–maize pathosystem has emerged as the current model for plant pathogenic basidiomycetes and as one of the few models for a true biotrophic interaction that persists throughout fungal development inside the host plant. This is based on the highly advanced genetic system for both the pathogen and its host, the ability to propagate U. maydis in axenic culture, and its unique capacity to induce prominent disease symptoms (tumors) on all aerial parts of maize within less than a week. The corn smut pathogen, though economically not threatening, will continue to serve as a model for related obligate biotrophic fungi such as the rusts, but also for closely related smut species that induce symptoms only in the flower organs of their hosts. In this review we describe the most prominent features of the U. maydis–maize pathosystem as well as genes and pathways most relevant to disease. We highlight recent developments that place this system at the forefront of understanding the function of secreted effectors in eukaryotic pathogens and describe the expected spin-offs for closely related species exploiting comparative genomics approaches.

The corn smut Ustilago maydis establishes a biotrophic interaction with its host plant maize. This interaction requires efficient suppression of plant immune responses, which is attributed to secreted effector proteins. Previously we identified Pep1 (Protein essential during penetration-1) as a secreted effector with an essential role for U. maydis virulence. pep1 deletion mutants induce strong defense responses leading to an early block in pathogenic development of the fungus. Using cytological and functional assays we show that Pep1 functions as an inhibitor of plant peroxidases. At sites of Δpep1 mutant penetrations, H₂O₂ strongly accumulated in the cell walls, coinciding with a transcriptional induction of the secreted maize peroxidase POX12. Pep1 protein effectively inhibited the peroxidase driven oxidative burst and thereby suppresses the early immune responses of maize. Moreover, Pep1 directly inhibits peroxidases in vitro in a concentration-dependent manner. Using fluorescence complementation assays, we observed a direct interaction of Pep1 and the maize peroxidase POX12 in vivo. Functional relevance of this interaction was demonstrated by partial complementation of the Δpep1 mutant defect by virus induced gene silencing of maize POX12. We conclude that Pep1 acts as a potent suppressor of early plant defenses by inhibition of peroxidase activity. Thus, it represents a novel strategy for establishing a biotrophic interaction. The maize pathogen U. maydis establishes a biotrophic interaction with its host plant and causes the formation of plant tumors. The U. maydis infection is initiated by a direct penetration of the plant epidermis and relies on living plant tissue. Therefore, suppression of the host immune system is essential for successful infection. Previously we identified the secreted effector Pep1, which is essential for U. maydis pathogenicity. pep1 deletion mutants are blocked by host defense responses immediately upon penetration. In the present study we identified the molecular function of Pep1 and explain its crucial role for fungal virulence. We found that Pep1 inhibits the plant oxidative burst, which is characterized by the accumulation of reactive oxygen species (ROS) such as hydrogen peroxide. A conserved component of the plant ROS generating system are peroxidases. We could show that Pep1 directly inhibits plant peroxidases. One specific maize peroxidase (POX12), which was strongly induced by infection of the pep1 deletion, directly interacts with POX12 in vivo. Moreover, POX12 silenced plants are penetrated by the pep1 deletion mutant, indicating functional relevance of the Pep1-POX12 interaction. Together, these findings show that Pep1 directly interferes with the ROS-generating system of the host plant to suppress immune responses.

Fungi are among the dominant causal agents of plant diseases. To colonize plants and cause disease, pathogenic fungi use diverse strategies. Some fungi kill their hosts and feed on dead material (necrotrophs), while others colonize the living tissue (biotrophs). For successful invasion of plant organs, pathogenic development is tightly regulated and specialized infection structures are formed. To further colonize hosts and establish disease, fungal pathogens deploy a plethora of virulence factors. Depending on the infection strategy, virulence factors perform different functions. While basically all pathogens interfere with primary plant defense, necrotrophs secrete toxins to kill plant tissue. In contrast, biotrophs utilize effector molecules to suppress plant cell death and manipulate plant metabolism in favor of the pathogen. This article provides an overview of plant pathogenic fungal species and the strategies they use to cause disease.

The basidiomycete Ustilago maydis causes smut disease in maize. Colonization of the host plant is initiated by direct penetration of cuticle and cell wall of maize epidermis cells. The invading hyphae are surrounded by the plant plasma membrane and proliferate within the plant tissue. We identified a novel secreted protein, termed Pep1, that is essential for penetration. Disruption mutants of pep1 are not affected in saprophytic growth and develop normal infection structures. However, Δpep1 mutants arrest during penetration of the epidermal cell and elicit a strong plant defense response. Using Affymetrix maize arrays, we identified 116 plant genes which are differentially regulated in Δpep1 compared to wild type infections. Most of these genes are related to plant defense. By in vivo immunolocalization, live-cell imaging and plasmolysis approaches, we detected Pep1 in the apoplastic space as well as its accumulation at sites of cell-to-cell passages. Site-directed mutagenesis identified two of the four cysteine residues in Pep1 as essential for function, suggesting that the formation of disulfide bridges is crucial for proper protein folding. The barley covered smut fungus Ustilago hordei contains an ortholog of pep1 which is needed for penetration of barley and which is able to complement the U. maydis Δpep1 mutant. Based on these results, we conclude that Pep1 has a conserved function essential for establishing compatibility that is not restricted to the U. maydis / maize interaction. For many fungi that infect plants, successful invasion is coupled to a series of differentiation steps that are necessary to breach the plant cuticle. Such fungi form specialized infection structures which allow direct penetration of the plant cuticle. The smut fungus Ustilago maydis establishes a biotrophic interaction with its host plant maize in which the infected host cells stay alive. During biotrophic growth, the intracellularly growing hyphae are encased by the host plasma membrane. We show here that a small effector protein, which is secreted by fungal hyphae during penetration, is absolutely essential for fungal entry into plant cells. When this effector is absent, hyphal cells penetrate the plant cell wall and invagination of the plant plasma membrane is observed, but any further fungal development is arrested. This arrest coincides with the induction of massive plant defense responses. Thus, this effector, which is conserved in related fungal species, plays an essential role in suppression of plant defense responses and is critical for establishing compatibility. This is the first example where a single effector protein assumes such a crucial role for infection-related development in a plant pathogenic fungus.

From Blight to Powdery Mildew: Pathogenic effects of microbes on plants have widespread consequences. Witness, for example, the cultural upheavals driven by potato blight in the 1800s. A variety of microbial pathogens continue to afflict crop plants today, driving both loss of yield and incurring the increased costs of control mechanisms. Now, four reports analyze microbial genomes in order to understand better how plant pathogens function (see the Perspective by Dodds ). Raffaele et al. (p. 1540 ) describe how the genome of the potato blight pathogen accommodates transfer to different hosts. Spanu et al. (p. 1543 ) analyze what it takes to be an obligate biotroph in barley powdery mildew, and Baxter et al. (p. 1549 ) ask a similar question for a natural pathogen of Arabidopsis . Schirawski et al. (p. 1546 ) compared genomes of maize pathogens to identify virulence determinants. Better knowledge of what in a genome makes a pathogen efficient and deadly is likely to be useful for improving agricultural crop management and breeding.

Summary: The fungal pathogen Ustilago maydis establishes a biotrophic relationship with its host plant maize (Zea mays). Hallmarks of the disease are large plant tumours in which fungal proliferation occurs. Previous studies suggested that classical defence pathways are not activated. Confocal microscopy, global expression profiling and metabolic profiling now shows that U. maydis is recognized early and triggers defence responses. Many of these early response genes are downregulated at later time points, whereas several genes associated with suppression of cell death are induced. The interplay between fungus and host involves changes in hormone signalling, induction of antioxidant and secondary metabolism, as well as the prevention of source leaf establishment. Our data provide novel insights into the complexity of a biotrophic interaction.