First Report of Melon Severe Mosaic Orthotospovirus infecting cucurbits in the United States.

Cucurbits (family Cucurbitaceae) includes globally important fruit and vegetable crops. Virus diseases pose a serious threat to cucurbits, limiting crop quality and yield (Regina et al. 2021). In fall 2023, leaf and fruit samples from two squash plants with chlorotic mosaic symptoms and fruit distortion from Monroe and Pope counties in Arkansas were received for diagnosis at the University of Arkansas Division of Agriculture Plant Clinic. Based on symptoms, samples were assessed for melon severe mosaic orthotospovirus (MeSMV) using the ImmunoStrip® developed for detection of the virus (Agdia® Inc., Elkhart, Indiana). The presence of MeSMV was also confirmed by RT-PCR using the Agdia Tospovirus group PCR primers. An amplicon was sequenced and showed 91% sequence identity to the MESMV type isolate (NC_033834, VE440-A). To further verify the results, nucleic acids from a squash sample from Pope County were extracted as described by Poudel et al. (2013), DNase treated, and sequenced on an Oxford Nanopore MinION as described by Liefting et al. (2021). A total of 25,914 raw reads were analyzed using VirFind (Ho and Tzanetakis 2014), which identified 112 reads mapping to the three segments of MeSMV. Primers for all three RNAs were developed and amplified 638, 650, and 1153 nt of the S, M, and L segments of the virus respectively. The amplicons were sequenced bidirectionally and show 89-93% identity to the type isolate from Mexico (GenBank accessions PP301332-4). MeSMV has only been identified in Mexico and can cause significant losses to honeydew melon, zucchini, and cucumber (Ciuffo et al. 2009). Thus, this is the first report of MeSMV outside Mexico. Given the severity of the symptoms observed in cucurbit crops, the virus poses a potential threat to the cucurbit industry in the United States. Growers should be aware of this virus and take the necessary precautions to prevent its spread in the field.