First report of a rust fungus (Puccinia sp.) infecting lemongrass in Minnesota.

In July 2021 and July - Oct. 2022, in a community garden near Mankato, Minnesota, rust disease was observed on lemongrass (Cymbopogon citriatus). In 2022, all 20 plants in the garden plot were infected. Lemongrass is used in some Asian cuisines and for tea or medicine. It is not hardy in Minnesota but is grown in gardens and outdoors in small-scale production. Uredinia were cinnamon-brown on the abaxial surface of leaf blades. Pustules were small (0.2 - 0.5 x .01 - .05 mm) and numerous, causing large necrotic lesions and leaf dieback (Fig. 1A). Severity ranged from 5 - 50% leaf loss. Urediniospores were finely echinulate, slightly ovular (22-25 x 20-23 μm), thick-walled (2.5-4 μm), with 3-4 roughly equatorial, sometimes scattered germ pores (Fig 1B; 1C). Clavate paraphyses were abundant. Other spore types were not observed. The pycnidia of a mycoparasitic fungus were present within the uredinia. The specimen was submitted to the Arthur Fungarium at Purdue University (PUR N24011). Primers ITS1rustF10d (Barnes and Szabo, 2007) and ITSRu1 (Rioux et al., 2015) were used to generate amplicons for the rust fungus, and ITS4 and ITSF+ (White, 1990) for the mycoparasite. Amplicons were sequenced on an Oxford Nanopore MinIon with R9 flow cells following manufacturer instructions. Reads (PRJNA802078) were filtered for quality (> Q13) and length (> 200 bp), mapped to reference sequences, aligned, and separated based on similarity. Consensus sequences were generated for the amplicons of the rust fungus and of two other fungi. BLASTn searches of the ITS sequences, OM442990 and OM442991, identified an Alternaria sp. (99.8% match (597/598) with MT548677) and Sphaerellopsis filum (syn. Darluca filum; 98.3% (529/538 bp) match with EF600974), a common rust mycoparasite. A BLASTn search of the rust fungal ITS sequence (OM442989) yielded 98.9% (549/555) and 98.6% (633/642) match with MT955206 and MT955207, respectively, both Puccinia cesatii on Bothriochloa ischaemum. The third closest match is P. cymbopogonis on C. citriatus (97.1% (595/613) with KY764115). Urediniospore morphology is consistent with that of P. cesatii (Cummins, 1971). Available evidence suggests the fungus is P. cesatii or a closely related species. Puccinia cesatii has been reported infecting Cymbopogon spp. (Stevenson et al., 1926; Dhar and Rekha, 1984), but lemongrass is not generally considered a host-possibly due to confusion of P. cesatii with P. cymbopogonis, a closely related rust pathogen of lemongrass that is morphologically very similar to P. cesatii. P. cymbopogonis has not been reported in the U.S. Rust diseases of lemongrass have been reported in three states: Hawaii (Gardner, 1985), California (Koike and Molinar, 1999), and Florida (Ploetz et al., 2014). In each case, the rust was identified as Puccinia nakanishikii. Urediniospores of P. nakanishikii are larger (26-36 μm long) (Cummins, 1971) and the ITS2 has no significant sequence similarity. P. cesatii is widespread in Eurasia, the southwest U.S., and Mexico (Cummins, 1971). Cummins lists three genera closely related to Cymbopogon as telial hosts of P. cesatii: Bothriochloa, Capillipedium, and Dicanthium. He lists nine rust fungi that infect Cymbopogon but does not list P. cesatii. Of these nine species, only P. cymbopogonis is morphologically similar. Further research is needed to investigate the potential impact of rust fungi on lemongrass production and to elucidate phylogenetic relationships of rust fungi infecting lemongrass.