The genus Phyllachora contains numerous obligate fungal parasites that produce raised, melanized structures called stromata on their plant hosts. Most members of this genus are not of significant economic concern, with the exception of P. maydis, the causal agent of tar spot of maize (Zea mays). Tar spot of maize has emerged as a major threat to maize production throughout the Americas and continues to spread throughout North America. To date, species designations for Phyllachora have been based on host associations and morphology, and the origin and diversity of the pathogen that causes tar spot is unknown. We assessed the sequence diversity of 186 single stroma isolates collected from 16 hosts representing 15 countries by amplification of the ITS and LSU gene regions. Samples included both herbarium and contemporary strains that covered a temporal range from 1905-2019. These 186 isolates were grouped into 5 distinct species with strong bootstrap support. We found three closely related, but genetically distinct groups of Phyllachora are capable of infecting maize in the United States, we refer to these as the P. maydis species complex. Based on herbarium species, we hypothesize that these three groups in the P. maydis species complex originated from Central America, Mexico and the Caribbean. Although two of these groups were only found on maize, the third and largest group contained contemporary strains found on maize and other grass hosts, as well as herbarium specimens from maize and other grasses that include 10 species of Phyllachora. The herbarium specimens were identified based on morphology and host association, but our data indicates there may be significant synonymy in the Phyllachora genus and additional work on species delineation and host specificity should be considered.
