A secreted Ustilago maydis effector promotes virulence by targeting anthocyanin biosynthesis in maize

Abstract

The biotrophic fungus Ustilago maydis causes smut disease in maize with characteristic tumor formation and anthocyanin induction. Here, we show that anthocyanin biosynthesis is induced by the virulence promoting secreted effector protein Tin2. Tin2 protein functions inside plant cells where it interacts with maize protein kinase ZmTTK1. Tin2 masks a ubiquitin–proteasome degradation motif in ZmTTK1, thus stabilizing the active kinase. Active ZmTTK1 controls activation of genes in the anthocyanin biosynthesis pathway. Without Tin2, enhanced lignin biosynthesis is observed in infected tissue and vascular bundles show strong lignification. This is presumably limiting access of fungal hyphae to nutrients needed for massive proliferation. Consistent with this assertion, we observe that maize brown midrib mutants affected in lignin biosynthesis are hypersensitive to U. maydis infection. We speculate that Tin2 rewires metabolites into the anthocyanin pathway to lower their availability for other defense responses.The production of agricultural crop plants is severely hindered by bacteria, viruses, and fungi that have developed their own strategies to colonize these plants and obtain nutrients from them. Some pathogens kill the plants they colonize, but ‘biotrophic pathogens’ employ sophisticated strategies to manipulate the host plant without killing it.During the past decade it has been recognized that the interactions between plants and biotrophic pathogens are largely governed by effector proteins—which are typically secreted by the pathogen after it makes contact with the host. These effector proteins can either stay in the space between the plant cells and pathogen cells, or actually enter inside the plant cells.The fungus Ustilago maydis is one such biotrophic pathogen that colonizes maize plants and causes a disease called corn smut. Hallmarks of this infection are the formation of large plant tumors and the production of a red pigment, called anthocyanin, in infected plant tissues.Now, Tanaka et al. reveal that an effector called Tin2, which is secreted by the corn smut fungus, causes the production of this anthocyanin pigment. Tin2 moves inside plant cells, where it blocks the breakdown of a protein-modifying enzyme that is necessary to ‘switch on’ the production of anthocyanin. When a mutant fungus that lacks Tin2 infects a maize plant, no anthocyanin is induced and the pathogen fails to reach the vascular tissue, where it would normally get most of its nutrients. Tanaka et al. revealed that, in these infections, this vascular tissue was strongly reinforced with a compound, called lignin, suggesting that the plant fortifies these cell walls to block access by the mutant fungus.Since the building blocks for making lignin are also required for making anthocyanins, Tanaka et al. suggest a model whereby Tin2 compromises the ability of plants to protect themselves by diverting resources away from making lignin. In line with this speculation, the corn smut fungus was shown to cause stronger disease symptoms in maize plants with mutations that prevent them from producing lignin.The Tin2 effector of the corn smut fungus appears to target a critical protein in maize that can shift the balance of the plant’s metabolic pathways in favor of the pathogen. Further, since anthocyanin production is also observed after infections of plants with other microbes, these findings may have uncovered a microbial strategy to enhance virulence that is also employed by other plant pathogens.