An In-Depth Evaluation of Powdery Mildew Hosts Reveals One of the World's Most Common and Widespread Groups of Fungal Plant Pathogens.

Powdery mildews are highly destructive fungal plant pathogens that have a significant economic impact on both agricultural and ecological systems worldwide. The intricate relationship between powdery mildews and their host plants has led to cospeciation. In this study, we conducted an extensive evaluation of powdery mildew hosts to provide an updated understanding of the host ranges and distributions of these fungi. The "United States National Fungus Collections Fungus-Host Dataset" is the primary source of information for our analyses. The analysis of the dataset demonstrated the worldwide prevalence of powdery mildews; the data contained over 72,000 reports of powdery mildews, representing ∼8.7% of all host-fungal records. We have updated the taxonomy and nomenclature of powdery mildews. In total, powdery mildews infect ∼10,125 host taxa belonging to 205 families of flowering plants, which accounts for 1,970 genera in 200 countries across six continents. Furthermore, we estimate that powdery mildews infect approximately 2.9% of described angiosperm species. Our study underscores the need for regular updates on powdery mildew host information due to the continuously evolving taxonomy and the discovery of new host taxa. Since 1986, we estimate an additional 1,866 host taxa, 353 genera, and 36 families have been reported. Additionally, the identification of powdery mildew hosts provides valuable insights into the coevolutionary dynamics between the fungi and their plant hosts. Overall, this updated list provides valuable insights into the taxonomy and geographic distribution of powdery mildew species, which builds upon the previous work of Amano in 1986. Discerning the geographic spread and host range of economically significant plant pathogens is vital for biosecurity measures and identifying the origins and expansion of potentially harmful pathogens.

Extensive intragenomic variation in the internal transcribed spacer region of fungi.

Fungi are among the most biodiverse organisms in the world. Accurate species identification is imperative for studies on fungal ecology and evolution. The internal transcribed spacer (ITS) rDNA region has been widely accepted as the universal barcode for fungi. However, several recent studies have uncovered intragenomic sequence variation within the ITS in multiple fungal species. Here, we mined the genome of 2414 fungal species to determine the prevalence of intragenomic variation and found that the genomes of 641 species, about one-quarter of the 2414 species examined, contained multiple ITS copies. Of those 641 species, 419 (∼65%) contained variation among copies revealing that intragenomic variation is common in fungi. We proceeded to show how these copies could result in the erroneous description of hundreds of fungal species and skew studies evaluating environmental DNA (eDNA) especially when making diversity estimates. Additionally, many genomes were found to be contaminated, especially those of unculturable fungi.

First report of Curvularia leaf spot on Zea mays caused by Curvularia lunata in North Carolina.

In late July of 2022, corn leaves with spots were observed in a field at North Carolina State University's Vernon James Research and Extension Center (35.873294º N, 76.658599º W; Plymouth NC). The affected leaves contained small (two to three millimeters in diameter) tan-colored circular to oval lesions with reddish-brown margins and chlorotic haloes. The disease severity approached 50% on affected leaves. Leaf tissue from the margins of the lesions was excised and surface sterilized by soaking in 10 % bleach solution for two minutes followed by rinsing in distilled water. Next, leaf tissue was transferred to alkaline water agar amended with streptomycin (100 mgliter-1) and the plates were incubated at 27º C in dark. After one week, conidia were observed. Conidia were curved, light brown in color and contained three transverse septa. The size of the conidia (n= 30) ranged from 21-25 µm (average 24 µm) long by 7 to 10 µm wide (average 9 µm). These characteristics were typical of Curvularia lunata (Munkvold and White 2016). DNA from pure cultures was extracted, internal transcribed space region (ITS) was amplified using the primers ITS4 and ITS5 (White et al. 1990). Sequences were obtained and deposited into GenBank (accession no. OP998306). BLAST search indicated that the sequence was 100 % identical to C. lunata from Louisiana (MG 971305.1, Garcia- Aroca et al. 2018). Pathogenicity was confirmed on field corn cultivar 'Yellow dent' by inoculating two pots of two-week-old greenhouse-grown corn plants (6 plants per pot) by spraying conidial suspension with a concentration of 6 x 104 conidia /ml. The control plants (n= 6) were sprayed with distilled water and the plants were covered with a plastic bag for 24 hours. The plastic bags were removed and plants were moved to the greenhouse at 23º C. Four days after inoculation small round tan-colored lesions were observed and no symptoms on control plants. The experiment was repeated once and the pathogen was recovered from the symptomatic tissue. The morphology of the recovered isolate was similar to the original isolate. This is the first report of C. lunata causing leaf spot on corn in North Carolina. Although this disease has not been demonstrated to cause yield loss it has been detected more frequently in corn production areas of U.S. and should continue to be monitored going forward.

Identification and Prevalence of Seedborne Botrytis spp. in Dry Pea, Lentil, and Chickpea in Montana.

Botrytis spp. cause gray mold and are significant pathogens of pulse crops (dry pea, lentil, and chickpea). Seedling infection can result in plant stunting and death. In this study, 100 Botrytis isolates were recovered from pulse crop seed samples that were submitted to the Regional Pulse Crop Diagnostic Laboratory at Montana State University. Nine Botrytis spp. were found to be associated with pulse seeds in Montana based on a combination of cultural characteristics; the amplification of partial sequences of the G3PDH, HSP60, and RPB2 genes; and phylogenetic analysis. Botrytis cinerea (n = 54) was the predominant species, followed by B. euroamericana (n = 22) and B. prunorum (n = 11). There were a few isolates of B. mali and five novel Botrytis spp. that included one cryptic species. To determine the pathogenicity and aggressiveness of the isolates, dry pea cultivar Lifter, lentil cultivar Richlea, and chickpea cultivar Sierra, detached leaves were inoculated using mycelial plugs. Lesion diameter produced by Botrytis isolates on three hosts differed (P < 0.05). Aggressiveness of B. cinerea was high in all three hosts and varied among the tested isolates. Spore inoculations were conducted on greenhouse-grown dry pea, lentil and chickpea plants using one sporulating isolate each of B. cinerea, B. prunorum, and Botrytis sp. 1. Results indicated that these isolates were pathogenic on the tested hosts. This study illustrates that many species of Botrytis are associated with pulse crop seed in Montana and can be aggressive on multiple crops, which may have implications for disease management.

Fusarium spp. Associated With Root Rot of Pulse Crops and Their Cross-Pathogenicity to Cereal Crops in Montana.

Root rot caused by Fusarium species is a major problem in the pulse growing regions of Montana. Fusarium isolates (n = 112) were obtained from seeds and roots of chickpea, dry pea, and lentil. Isolates were identified by comparing the sequences of the internal transcribed spacer region and the translation elongation factor 1-α in Fusarium-ID database. Fusarium avenaceum was the most abundant species (28%), followed by F. acuminatum (21%), F. poae (13%), F. oxysporum (8%), F. culmorum (6%), F. redolens (6%), F. sporotrichioides (6%), F. solani (4%), F. graminearum (2%), F. torulosum (2%), and F. tricinctum (0.9%). The aggressiveness of a subset of 50 isolates that represent various sources of isolation was tested on three pulse crops and two cereal crops. Nonparametric analysis of variance conducted on ranks of disease severity indicated that F. avenaceum and F. solani isolates were highly aggressive on pea and chickpea. In lentil, F. avenaceum and F. culmorum were highly aggressive. In barley, F. avenaceum, F. solani, F. culmorum, and F. graminearum were highly aggressive. In wheat, F. avenaceum, F. graminearum, and F. culmorum were highly aggressive. Two F. avenaceum isolates were highly aggressive across all the crops tested and found to be cross-pathogenic. One isolate of F. culmorum and an isolate of F. graminearum obtained from chickpea and lentil seed were highly aggressive on barley and wheat. The results indicate that multiple Fusarium spp. from seeds and roots can cause root rot on both pulse and cereal crops. Rotating these crops may still lead to an increase in inoculum levels, making crop rotation limited in efficacy as a disease management strategy.

Phylogeny and taxonomy of Podosphaera cerasi, sp. nov., and Podosphaera prunicola sensu lato.

Powdery mildew of Prunus spp. is a significant disease in most cherry growing regions of Washington, USA. Powdery mildews on Prunus virginiana and Pr. avium were previously assigned to Podosphaera clandestina s. lat. (= Po. oxyacanthae) or Po. prunicola. In this report, we confirm the presence of two distinct Podosphaera species on these hosts. Phylogenetic analyses of nuc rDNA sequences from the internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS) and 28S subunit confirmed the presence of two distinct species. A morphological comparison with type material of Po. prunicola and additional collections demonstrated that the powdery mildew on Pr. virginiana (including var. demissa and var. melanocarpa) is in fact Po. prunicola. The powdery mildew on Pr. avium is genetically, morphologically, and biologically distinct from Po. prunicola and is described here as the new species Po. cerasi. Cross-inoculation experiments confirmed that these two species are host specific. Podosphaera prunicola was unable to colonize Pr. avium, whereas Po. cerasi was unable to colonize Pr. virginiana. Morphological reexamination of numerous specimens identified as Po. prunicola on a broad range of Prunus species suggests that Po. prunicola is probably confined to species in Prunus subgen.Padus (= Prunus subgen. Cerasus sect. Laurocerasus, including sect. Padus), with Pr. virginiana as the principal host. Podosphaera cerasi occurs on hosts in Prunus subgen. Cerasus, and our work confirms a newly described species of powdery mildew on Pr. avium. This work also includes the first documented and genetically proven European record of Po. prunicola on Pr. serotina and its widespread occurrence in the United States.

Proteomic profiling of L-cysteine induced selenite resistance in Enterobacter sp. YSU.

BACKGROUND: Enterobacter sp. YSU is resistant to several different heavy metal salts, including selenite. A previous study using M-9 minimal medium showed that when the selenite concentration was 100,000 times higher than the sulfate concentration, selenite entered Escherichia coli cells using two pathways: a specific and a non-specific pathway. In the specific pathway, selenite entered the cells through a yet to be characterized channel dedicated for selenite. In the non-specific pathway, selenite entered the cells through a sulfate permease channel. Addition of L-cystine, an L-cysteine dimer, appeared to indirectly decrease selenite import into the cell through the non-specific pathway. However, it did not affect the level of selenite transport into the cell through the specific pathway. RESULTS: Growth curves using M-9 minimal medium containing 40 mM selenite and 1 mM sulfate showed that Enterobacter sp. YSU grew when L-cysteine was present but died when it was absent. Differential protein expression analysis by two dimensional gel electrophoresis showed that CysK was present in cultures containing selenite and lacking L-cysteine but absent in cultures containing both selenite and L-cysteine. Additional RT-PCR studies demonstrated that transcripts for the sulfate permease genes, cysA, cysT and cysW, were down-regulated in the presence of L-cysteine. CONCLUSION: L-cysteine appeared to confer selenite resistance upon Enterobacter sp. YSU by decreasing the level of selenite transport into the cell through the non-specific pathway.