The <i>Epichloë</i> <i>festucae</i> Antifungal Protein <i>Efe</i>-AfpA Is Also a Possible Effector Protein Required for the Interaction of the Fungus with its Host Grass <i>Festuca rubra</i> subsp. <i>rubra</i>

• Main Authors: Ruying Wang, Simin Luo, Bruce B. Clarke, Faith C. Belanger
• Format: Article
• Language: English
• Published: MDPI AG 2021-01-01
• Series: Microorganisms
• Subjects: antifungal protein; endophyte; strong creeping red fescue; symbiosis
• Online Access: https://www.mdpi.com/2076-2607/9/1/140

Strong creeping red fescue (<i>Festuca rubra </i>subsp. <i>rubra</i>) is a commercially important low-maintenance turfgrass and is often naturally infected with the fungal endophyte <i>Epichloë</i> <i>festucae</i>. Epichloë spp. are endophytes of several cool-season grass species, often conferring insect resistance to the grass hosts due to the production of toxic alkaloids. In addition to insect resistance, a unique feature of the strong creeping red fescue/E. festucae symbiosis is the endophyte-mediated disease resistance to the fungal pathogen Clarireedia jacksonii, the causal agent of dollar spot disease. Such disease resistance is not a general feature of other grass/<i> Epichloë</i> interactions. E. festucae isolates infecting red fescue have an antifungal protein gene Efe-afpA, whereas most other <i>Epichloë</i> spp. do not have a similar gene. The uniqueness of this gene suggests it may, therefore, be a component of the unique disease resistance seen in endophyte-infected red fescue. Here, we report the generation of CRISPR-Cas9 Efe-afpA gene knockouts with the goal of determining if absence of the protein in endophyte-infected Festuca rubra leads to disease susceptibility. However, it was not possible to infect plants with the knockout isolates, although infection was possible with the wild type E. festucae and with complemented isolates. This raises the interesting possibility that, in addition to having antifungal activity, the protein is required for the symbiotic interaction. The antifungal protein is a small secreted protein with high expression in planta relative to its expression in culture, all characteristics consistent with effector proteins. If Efe-AfpA is an effector protein it must be specific to certain interactions, since most <i>Epichloë</i> spp. do not have such a gene in their genomes.