BackgroundUbiquitination is a major post-translational protein modification that regulates essentially all cellular and physiological pathways in eukaryotes. The ubiquitination process typically involves three distinct classes of enzymes, ubiquitin-activating enzyme (E1), ubiquitin-conjugating enzyme (E2) and ubiquitin ligase (E3). To date, a comprehensive identification and analysis of core components comprising of the whole soybean (Glycine max) ubiquitin system (UBS) has not been reported.ResultsWe performed a systematic, genome-wide analysis of genes that encode core members of the soybean UBS in this study. A total of 1431 genes were identified with high confidence to encode putative soybean UBS components, including 4 genes encoding E1s, 71 genes that encode the E2s, and 1356 genes encoding the E3-related components. Among the E3-encoding genes, 760 encode RING-type E3s, 124 encode U-box domain-containing E3s, and 472 encode F-box proteins. To find out whether the identified soybean UBS genes encode active enzymes, a set of genes were randomly selected and the enzymatic activities of their recombinant proteins were tested. Thioester assays indicated proteins encoded by the soybean E1 gene GmUBA1 and the majority of selected E2 genes are active E1 or E2 enzymes, respectively. Meanwhile, most of the purified RING and U-box domain-containing proteins displayed E3 activity in the in vitro ubiquitination assay. In addition, 1034 of the identified soybean UBS genes were found to express in at least one of 14 soybean tissues examined and the transcript level of 338 soybean USB genes were significantly changed after abiotic or biotic (Fusarium oxysporum and Rhizobium strains) stress treatment. Finally, the expression level of a large number of the identified soybean UBS-related genes was found significantly altered after soybean cyst nematode (SCN) treatment, suggesting the soybean UBS potentially plays an important role in soybean immunity against SCN.ConclusionsOur findings indicate the presence of a large and diverse number of core UBS proteins in the soybean genome, which suggests that target-specific modification by ubiquitin is a complex and important part of cellular and physiological regulation in soybean. We also revealed certain members of the soybean UBS may be involved in immunity against soybean cyst nematode (SCN). This study sets up an essential foundation for further functional characterization of the soybean UBS in various physiological processes, such as host immunity against SCN.Electronic supplementary materialThe online version of this article (10.1186/s12870-018-1365-7) contains supplementary material, which is available to authorized users.

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Tar spot caused by the fungus Phyllachora maydis Maubl. is a significant foliar disease of corn (Zea mays L.). Threatening corn production across the Americas, this disease can reduce the quality of silage and grain yield (Rocco da Silva et al. 2021; Valle-Torres et al. 2020). Lesions caused by P. maydis are usually black, glossy, and raised stromata on the leaf surface and occasionally on the husk. (Liu 1973; Rocco da Silva et al. 2021). Samples consistent with tar spot of corn were collected between September and October of 2022 from 6 fields in Kansas, 23 in Nebraska, and 6 in South Dakota. One sample was selected from each of the three states for further microscopic evaluation and molecular analysis. Signs of the fungus were visually and microscopically confirmed in eight Nebraska counties in October 2021; however, in the 2021, season tar spot sings were not found in Kansas and South Dakota. In the 2022 season disease severity varied by location; some fields in Kansas had <1% incidence, whereas in South Dakota disease incidence approached 1-2%, and in Nebraska between <1-5%. Stromata were present on both green and senescing tissues. Morphological characteristics of the pathogen were similar and consistent with the description of P. maydis (Parbery 1967) from all examined leaves across all locations. Asexual spores (conidia) were produced in pycnidial fruiting bodies ranging in size 1.29 to 2.82 x 8.84 to 16.95 µm (n = 40, average 1.98 × 13.30 μm). The pycnidial fruiting bodies were often found adjacent to perithecia within the stromata. For molecular confirmation, stromata were aseptically removed from leaves collected at each location and DNA extracted using a phenol chloroform method. The internal transcribed spacer (ITS) regions of the ribosomal RNA gene were sequenced using ITS1/ITS4 universal primers (Larena et al. 1999). Amplicons were Sanger sequenced (Genewiz, Inc., South Plainfield, NJ), and a consensus sequence for each sample was deposited in GenBank: Kansas (OQ200487), Nebraska (OQ200488), and South Dakota (OQ200489). Using the BLASTn, sequences from Kansas, Nebraska and South Dakota showed 100% homology with 100% query cover to other P. maydis GenBank accessions (MG881848.1; OL342916.1; OL342915.1). Koch’s postulates were not performed given the obligate nature of the pathogen (Muller and Samuels 1984). This report documents the first confirmation of tar spot on corn in Kansas, Nebraska, and South Dakota (Great Plains).

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