First report of Puccinia recondita rust on native jewelweed (Impatiens capensis) in Pennsylvania.

Jewelweed (Impatiens spp., Balsaminaceae) is a common native annual plant within Pennsylvania wetland ecosystems, many of which are under threat from invasive non-native plants, and is an important wetland indicator plant (code FACW; facultative wetland). In May 2014, rust disease symptoms on native jewelweed (Impatiens capensis Meerb.) were observed within a small (0.1 ha) wet area in York County, southeastern Pennsylvania (39.9080648oN, -77.2472024oW). Rust symptoms were noted on most jewelweed plants within the wet area. Foliar symptoms included chlorosis and premature defoliation; infected stems were distorted. Infected leaves and stems contained orange, erumpent aecia with white fragmented peridia (Fig. 1). Symptomatic leaves and stem sections were collected from five infected plants within one small (5 X 5 m) plot in the center of the wet area and taken to the laboratory for microscopic observations and morphological measurements. Mean aecia diameter was 299.2 ± 55.0 µm (n = 60). Aeciospores were single-celled, orange, and generally globose (Fig. 2) with a mean diameter of 24.4 ± 1.4 µm, (n = 60). Disease symptomology and aecia morphology were consistent with Puccinia recondita Dietel & Holw. DNA extraction (from infected stem material), polymerase chain reactions, and DNA sequencing of the 28S region of the nuclear ribosomal DNA repeat was conducted following protocols in Aime (2006) and Aime et al. (2018). The sequence shares 99.34% identity (903 / 909 bp) with P. recondita (BPI 910319) collected in California (KY798399). A voucher specimen has been deposited in the Arthur Fungarium at Purdue University (PUR N24229) with corresponding 28S sequence (GenBank accession OR648406). P. recondita has been reported on native I. capensis in Indiana (Koslow and Clay 2010) and North Carolina (Grand 1985), but not in Pennsylvania to the best of our knowledge (Farr and Rossman 2022). If this rust disease becomes severe on native jewelweeds in Pennsylvania, it may adversely affect our ability to accurately classify native wetlands in the state. In addition, P. recondita is a heteroecious rust that is a major pathogen of grain crops of economic importance (i.e., wheat, barley, oats), which are grown in southeastern Pennsylvania near the infested area. Further research is warranted to understand if native, annual jewelweed can serve as a secondary or alternate host to cause rust disease in major cereal crops.

First Report of Puccinia lagenophorae on Senecio vulgaris in Pennsylvania.

Common groundsel (Senecio vulgaris L.), is an aster native to Eurasia and is now a common weed in gardens, roadsides and vacant lots worldwide. In 2001, Scholler and Toike were first to report that common groundsel was a host for the rust fungus Puccinia lagenophorae Cooke in North America (Scholler and Toike 2001). This report from California was followed by reports of P. lagenophorae infections on common groundsel in New York, Oklahoma, and Oregon (Little-field et al. 2005). In 2007, Bruckart et al. published the first report of this host-pathogen combi-nation in Canada (Bruckart et al. 2007). To our knowledge, there are no published reports of P. lagenophorae on common groundsel in Pennsylvania (Farr and Rossman 2022). In May 2022, symptomatic common groundsel plants were observed in Biglerville, Adams Co., southern Penn-sylvania (N 39.9268047, E 77.2473878). Host plants exhibited conspicuous aecia on deformed stems (Fig. 1). Disease symptomology and morphology were consistent with P. langenophorae (Scholler and Toike 2001). P. lagenophorae is an autoecious rust that forms aecia and telia, but only aecia are typically formed on Senecio spp.; telia were not noted during our observations. Aecia were orange, cup-shaped, bordered by fragmented recurved peridia (Fig. 1), and they had mean diameters 262.9 ± 20.9 X 175.2 ± 22.2 µm (n = 20). Aeciospores were orange, oval, with mean diameters of 16.5 ± 0.97 X 11.5 ± 1.08 µm (n = 20). Fungal DNA was extracted from symptomatic stems. Polymerase chain reaction and sequencing of the 28S region of the nuclear ribosomal DNA repeat were conducted with primers Rust2inv and LR6 following protocols in Aime (2006). The sequence shared 100% identity (909 / 909 bp) with 8 sequences of P. la-genphorae in GenBank, including one on Ozothamnus cordatus from Perth, Western Australia, Australia (KF690699), vouchered in the Queensland Plant Pathology Herbarium (BRIP 57770), Brisbane, Queensland, Australia. A voucher specimen has been preserved in the Arthur Fungari-um at Purdue University (PUR N24039) with corresponding 28S sequence (GenBank accession OP718536).

Morphology, Ultrastructure, and Molecular Phylogeny of Rozella multimorpha, a New Species in Cryptomycota.

Increasing numbers of sequences of basal fungi from environmental DNA studies are being deposited in public databases. Many of these sequences remain unclassified below the phylum level because sequence information from identified species is sparse. Lack of basic biological knowledge due to a dearth of identified species is extreme in Cryptomycota, a new phylum widespread in the environment and phylogenetically basal within the fungal lineage. Consequently, we are attempting to fill gaps in the knowledge of Rozella, the best-known genus in this lineage. Rozella is a genus of unwalled, holocarpic, endobiotic parasites of hosts including Chytridiomycota, Blastocladiomycota, Oomycota, Basidiomycota, and a green alga, with most species descriptions based on morphology and host specificity. We found a Rozella parasitizing a Pythium host that was a saprobe on spruce pollen bait placed with an aquatic sample. We characterized the parasite with light microscopy, TEM of its zoospores and sporangia, and its 18S/28S rDNA. Comparison with other Rozella species indicates that the new isolate differs morphologically, ultrastructurally, and genetically from Rozella species for which we have data. Features of the zoospore also differ from those of previously studied species. Herein we describe the Rozella as a new species, R. multimorpha.

Wood decay fungus Flavodon ambrosius (Basidiomycota: Polyporales) is widely farmed by two genera of ambrosia beetles.

The ambrosia fungus Flavodon ambrosius is the primary nutritional mutualist of ambrosia beetles Ambrosiodmus and Ambrosiophilus in North America. F. ambrosius is the only known ambrosial basidiomycete, unique in its efficient lignocellulose degradation. F. ambrosius is associated with both native American beetle species and species introduced from Asia. It remains unknown whether F. ambrosius is strictly a North American fungus, or whether it is also associated with these ambrosia beetle genera on other continents. We isolated fungi from the mycangia and galleries of ambrosia beetles Ambrosiodmus rubricollis, Ambrosiodmus minor, Ambrosiophilus atratus, and Ambrosiophilus subnepotulus in China, South Korea, and Vietnam. Phylogenetic analyses suggest that all Asian and North American isolates represent a single haplotype. These results confirm Flavodon ambrosius as the exclusive mutualistic fungus of multiple Ambrosiodmus and Ambrosiophilus beetle species around the world, making it the most widespread known ambrosia fungus species, both geographically and in terms of the number of beetle species. The Flavodon-beetle symbiosis appears to employ an unusually strict mechanism for maintaining fidelity, compared to the symbioses of the related Xyleborini beetles, which mostly vector more dynamic fungal communities.

New Fungus-Insect Symbiosis: Culturing, Molecular, and Histological Methods Determine Saprophytic Polyporales Mutualists of Ambrosiodmus Ambrosia Beetles.

Ambrosia symbiosis is an obligate, farming-like mutualism between wood-boring beetles and fungi. It evolved at least 11 times and includes many notorious invasive pests. All ambrosia beetles studied to date cultivate ascomycotan fungi: early colonizers of recently killed trees with poor wood digestion. Beetles in the widespread genus Ambrosiodmus, however, colonize decayed wood. We characterized the mycosymbionts of three Ambrosiodmus species using quantitative culturing, high-throughput metabarcoding, and histology. We determined the fungi to be within the Polyporales, closely related to Flavodon flavus. Culture-independent sequencing of Ambrosiodmus minor mycangia revealed a single operational taxonomic unit identical to the sequences from the cultured Flavodon. Histological sectioning confirmed that Ambrosiodmus possessed preoral mycangia containing dimitic hyphae similar to cultured F. cf. flavus. The Ambrosiodmus-Flavodon symbiosis is unique in several aspects: it is the first reported association between an ambrosia beetle and a basidiomycotan fungus; the mycosymbiont grows as hyphae in the mycangia, not as budding pseudo-mycelium; and the mycosymbiont is a white-rot saprophyte rather than an early colonizer: a previously undocumented wood borer niche. Few fungi are capable of turning rotten wood into complete animal nutrition. Several thousand beetle-fungus symbioses remain unstudied and promise unknown and unexpected mycological diversity and enzymatic innovations.