First report of Erysiphe cruciferarum causing powdery mildew of Alliaria petiolata in Maryland.

Alliaria petiolata (Bieb.) Cavara & Grande (garlic mustard) is a biennial crucifer native to Europe and invasive in North America, where it outcompetes native plants in deciduous forests. In July 2021, powdery mildew was observed on A. petiolata in Frederick County, Maryland. Signs of the disease included white, tomentose mycelium producing abundant conidia (Fig S1). A majority of plants were affected, and severity ranged from the presence of small, discrete infections to complete colonization of leaves, stems, and ripening seed pods. Conidia from field collected leaves were transferred to disease-free A. petiolata for maintenance in a growth chamber at 20°C and 80% RH with a 12 hr photoperiod. Fungal morphology was recorded 30 days after this transfer. Appressoria were irregularly lobed, and conidiophores were straight and composed of 2-3 cells. Cylindrical to oblong conidia were produced singly in pseudochains of 2-6 (x = 3), measured 39-64 by 18-29 (x = 52 by 24) μm, had a length to width ratio greater than two, and germinated at the ends. Fibrosin bodies were absent from conidia, and chasmothecia were not observed in the field or on inoculated plant material. Based on anamorphic characteristics, the pathogen was placed in the genus Erysiphe (Boesewinkel 1980). Species level identity was determined using DNA sequences. Conidia and mycelia were scraped from leaves and used for genomic DNA extraction with the Quick-DNA Fungal/Bacterial Miniprep Kit (Zymo Research, Irvine, CA). A portion of the internal transcribed spacer region of rDNA was amplified using the primers ITS5/ITS4 (White et al. 1990). Purified amplicons (PCR & DNA Cleanup Kit, New England BioLabs Inc., Ipswich, MA) were sequenced at Eurofins Genomics (Louisville, KY). The resulting sequence was compared to those in NCBI GenBank using the blastn algorithm (Altschul et al. 1990). The newly generated sequence (GenBank: OK157430) was identical (599/599 bp) to samples of E. cruciferarum from the United Kingdom (GenBank: KY660931.1, KY660879.1, KY660752.1). Because E. cruciferarum sensu lato is heterogeneous (Pastircakova et al. 2016), additional sequence comparisons were made to the E. cruciferarum sensu stricto holotype (589/599 bp, GenBank: KU672364) and a vouchered E. cruciferarum s. lat. (596/599 bp, GenBank: LC009943). This supports identification of the pathogen as E. cruciferarum s. lat. and suggests the taxonomy of isolates from A. petiolata should be reassessed following any revision to E. cruciferarum. A modified Koch's postulates procedure was followed to confirm pathogenicity. Leaves colonized by E. cruciferarum were briefly pressed against the leaves of three disease-free plants grown from seed in a greenhouse. After 14 days, inoculated plants showed signs of powdery mildew similar to those observed in the field, and a control treatment using pathogen-free leaves resulted in no disease. This inoculation experiment was performed twice, and the identity of the pathogen was reconfirmed based on morphology. This is the first report of powdery mildew on A. petiolata in Maryland. Erysiphe cruciferarum s. lat. is widely distributed on other hosts and has been found on A. petiolata throughout Europe and in Ohio and Indiana (Farr and Rossman 2021; Blossey et al. 2001; Enright and Cipollini 2007; Ciola and Cipollini 2011). This pathogen has been proposed as a biological control agent (Cipollini and Enright 2009; Cipollini et al. 2020), and the presence of disease in Maryland suggests the local population of A. petiolata is susceptible to E. cruciferarum and the environment there is favorable to disease development. References: Altschul, S. F., et al. 1990. J. Mol. Biol. 215:403. Blossey, B., et al. 2001. Nat. Areas J. 21:357. Boesewinkel, H. J. 1980. Bot. Rev. 46:167. Ciola, V., and Cipollini, D. 2011. Am. Midl. Nat. 166:40-52. Cipollini, D., and Enright, S. M. 2009. Invasive Plant Sci. Manag. 2:253. Cipollini, D., et al. 2020. Biol. Invasions. 22:1657-1668. Enright, S. M., and Cipollini, D. 2007. Am. J. Bot. 94:1813. Farr, D. F., and Rossman, A. Y. 2021. Fungal Databases, Syst. Mycol. Microbial. Lab., ARS, USDA. https://nt.ars-grin.gov/fungaldatabases/ Pastircakova, K., et al. 2016. Mycol. Prog. 15:36 White, T. J., et al. 1990. Page 315 in: PCR Protocols. A Guide to Methods and Applications, Academic Press, San Diego. Keywords: Alliaria petiolata, Erysiphe cruciferarum, garlic mustard, powdery mildew, invasive plant, biocontrolFunding and Disclaimer: The author(s) declare no conflict of interest. This work was supported by USDA-ARS Appropriated Project Number 8044-22000-047-000-D. Mention of trade names or commercial products in this report is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. USDA is an equal opportunity lender, provider, and employer.