First Report of Beech Leaf Disease, Caused by Litylenchus crenatae mccannii, on American Beech (Fagus grandifolia) in Virginia.

Beech leaf disease (BLD) was first reported in 2012 in Lake County, Ohio on American beech trees (Fagus grandifolia Ehrh.). Since then, it spread across the Northeastern United States and has been reported from Ohio, Pennsylvania, New York, New Jersey, Connecticut, Rhode Island, Maine, West Virginia, and Ontario, Canada (Carta et al. 2020; Mara and LaMondia 2020, Reid et al. 2020). The symptoms of BLD are characterized by dark interveinal banding of leaves appearing soon after spring flush that become chlorotic and necrotic through autumn, resulting in canopy thinning in advanced stages, followed in some young trees by death. Litylenchus crenatae mccannii has similar morphological characteristics with Litylenchus crenatae (Kanzaki et al. 2019) reported on Fagus crenata from Japan. However that beech species has not shown BLD symptoms or yielded any L. crenatae mccannii in North America. There are several morphological differences between the two. The North American subspecies have shorter post-uterine sac, narrower body width in mature females, shorter tail in immature females, longer tail in mature females, and longer stylet in males when compared to the Japanese subspecies (Carta et al. 2020). BLD symptoms were found on American beech trees in Prince William Forest Park, Prince William County, Virginia in June, 2021. The affected leaves contained females, males, and juveniles with morphometrics consistent with L. crenatae mccannii (Carta et al. 2020). The crude genomic DNA from a live single Litylenchus was prepared with freeze-thaw lysis (Carta and Li, 2019). The ITS PCR were performed by using the procedures and primer set, ITS-CL-F2 and 28S-CL-R described in the previous study (Carta and Li, 2020). The visualization, the cleanup and the direct DNA sequencing of the PCR products were performed by using the procedures described in the previous studies (Carta and Li, 2018 and 2019). Sequences were submitted to GenBank as accessions MZ611855 and MZ611856. This represents the first report of BLD in Virginia. It is also approximately 300 miles south of the 2020 detection of BLD from New Cumberland, WV, and represents the southernmost detection of the disease and nematode in North America. The author(s) declare no conflict of interest. References Carta, L.K., Li, S. 2018. Improved 18S small subunit rDNA primers for problematic nematode amplification. Journal of Nematology. 50, 533-542. Carta, L.K., Li, S. 2019. PCR amplification of a long rDNA segment with one primer pair in agriculturally important nematodes. Journal of Nematology. 51, e2019-26. Carta, L.K., Li, S. 2020. Improvement of long segment ribosomal PCR amplification for the molecular taxonomic identification of Litylenchus crenatae mccannii in beech trees with beech leaf disease. Journal of Nematology. 52, e2020-016. Kanzaki, N., Ichihara, Y., Aikawa, T., Ekino, T., Masuya, H. 2019. Litylenchus crenatae n. sp. (Tylenchomorpha: Anguinidae), a leaf gall nematode parasitising Fagus crenata Blume Nematology 21 (1), 5-22. http://www.brill.com/nematology doi: 10.1163/15685411-00003190 Marra, R.E., LaMondia, J. 2020. First report of beech leaf disease, caused by the foliar nematode, Litylenchus crenatae mccannii, on American beech (Fagus grandifolia) in Connecticut. Plant Disease (early view). https://doi.org/10.1094/PDIS-02-20-0442-PDN Reed, S. E., Greifenhagen, S., Yu, Q., Hoke A., Burke D. J., Carta L. K., Handoo Z.A., Kantor, M.R., Koch, J. 2020. Foliar nematode, Litylenchus crenatae ssp. mccannii, population dynamics in leaves and buds of beech leaf disease-affected trees in Canada and the US. Forest Pathology 50 (3), e12599.