Identification of Neopestalotiopsis spp. from Strawberry Leaf, Fruit, and Crown Tissues in North Carolina

North Carolina is a leading fresh-market strawberry producer in the southeastern United States, with increasing cultivation driven by consumer demand. In recent years, Neopestalotiopsis-associated diseases have emerged as a major threat to strawberry production, yet limited information is available on their distribution, species diversity, and pathogenic variation in the region. This study investigated the occurrence and characterization of <i>Neopestalotiopsis</i> species associated with strawberry crown rot samples submitted to the North Carolina State University Plant Disease and Insect Clinic during 2023-2024. Crowns of diseased plants collected from 27 counties, representing 17 cultivars, were cultured. <i>Neopestalotiopsis</i> was the predominant genus (<i>n</i> = 114), represented by <i>N. hispanica</i> (<i>n</i> = 67), species from the <i>N. rosae</i> complex (<i>n</i> = 44), <i>N. clavispora</i> (<i>n</i> = 1), <i>N. scalabiensis</i> (<i>n</i> = 1), and <i>N. longiappendiculata</i> (<i>n</i> = 1). Greenhouse pathogenicity assays confirmed that the tested <i>Neopestalotiopsis</i> isolates were able to cause disease on the strawberry cultivar 'Fresca'. These findings provide the first comprehensive overview of <i>Neopestalotiopsis</i> species associated with strawberry crown rot in North Carolina and highlight their genetic and pathogenic diversity, contributing to improved understanding and management of this emerging disease.