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Heterodera glycines, the soybean cyst nematode (SCN), and fungal pathogen Macrophomina phaseolina are economically important soybean pathogens that may coinfest fields. Resistance remains the most effective management tactic for SCN, and the rhg1-b resistance allele derived from plant introduction 88788 is most commonly deployed in the northern United States. The concomitant effects of SCN and M. phaseolina on soybean performance, as well as the effect of the rhg1-b allele in two different genetic backgrounds, were evaluated in three environments (during 2013 to 2015) and a greenhouse bioassay. Within two soybean populations, half of the lines had the rhg1-b allele, and the other half had the susceptible allele in the backgrounds of the cultivars IA3023 and LD00-3309. Significant interactions between soybean rhg1-b allele and M. phaseolina-infested plots were observed in 2014. In all experiments, initial SCN populations (Pi) and M. phaseolina in roots were associated with reduced soybean yield. SCN reproduction factor (RF = final population/Pi) was affected by SCN Pi, rhg1-b, and genetic background. A background-by-genotype interaction on yield was observed only in 2015, with a stronger rhg1-b effect in the LD00-3309 background, which suggested that the susceptible parent ‘IA3023’ is tolerant to SCN. SCN female index from greenhouse experiments was compared with field RF, and Lin’s concordance and Pearson’s correlation coefficients decreased with increasing field SCN Pi in soil. In this study, both SCN and M. phaseolina reduced soybean yield asymptomatically, and the impact of SCN rhg1-b resistance was dependent on SCN virulence but also population density.

Species of Phytophthora, Phytopythium, and Pythium affect soybean seed and seedlings each year, primarily through reduced plant populations and yield. Oxathiapiprolin is effective at managing several foliar diseases caused by some oomycetes. The objectives of these studies were to evaluate oxathiapiprolin in a discriminatory dose assay in vitro; evaluate oxathiapiprolin as a soybean seed treatment on a moderately susceptible cultivar in 10 environments; compare the impact of seed treatment on plant populations and yields in environments with low and high precipitation; and compare a seed treatment mixture on cultivars with different levels of resistance in four environments. There was no reduction in growth in vitro among 13 species of Pythium at 0.1 µg ml−1. Soybean seed treated with the base fungicide plus oxathiapiprolin (12 and 24 µg a.i. seed−1) alone, oxathiapiprolin (12 µg a.i. seed−1) plus mefenoxam (6 µg a.i. seed−1), or oxathiapiprolin (24 µg a.i. seed−1) plus ethaboxam (12.1 µg a.i. seed−1) had greater yields in environments that received ≥50 mm of precipitation within 14 days after planting compared with those that received less. Early plant population and yield were significantly higher for seed treated with oxathiapiprolin (24 µg a.i. seed−1) + metalaxyl (13.2 µg a.i. seed−1) compared with nontreated for six of seven cultivars in at least one of four environments. Oxathiapiprolin combined with another Oomycota fungicide applied to seed has the potential to be used to protect soybean plant establishment and yield in regions prone to poor drainage after high levels of precipitation.

Phytophthora sojae, the causal agent of Phytophthora root and stem rot of soybean, has been managed with single Rps genes since the 1960s but has subsequently adapted to many of these resistance genes, rendering them ineffective. The objective of this study was to examine the pathotype and genetic diversity of P. sojae from soil samples across Illinois, Indiana, Kentucky, and Ohio by assessing which Rps genes were still effective and identifying possible population clusters. There were 218 pathotypes identified from 473 P. sojae isolates with an average of 6.7 out of 15 differential soybean lines exhibiting a susceptible response for each isolate. Genetic characterization of 103 P. sojae isolates from across Illinois, Indiana, Kentucky, and Ohio with 19 simple sequence repeat markers identified 92 multilocus genotypes. There was a moderate level of population differentiation between these four states, with pairwise FSTvalues ranging from 0.026 to 0.246. There were also moderate to high levels of differentiation between fields, with pairwise FSTvalues ranging from 0.071 to 0.537. Additionally, cluster analysis detected the presence of P. sojae population structure across neighboring states. The level of pathotype and genetic diversity, in addition to the identification of population clusters, supports the hypothesis of occasional outcrossing events that allow an increase in diversity and the potential to select for a loss in avirulence to specific resistance genes within regions. The trend of suspected gene flow among neighboring fields is expected to be an ongoing issue with current agricultural practices.

The Soybean Nested Association Mapping (SoyNAM) populations were developed from 40 parents and have been used to map genes underlying complex traits such as yield and disease resistance. Soil borne pathogens that affect soybean seed and seedlings result in significant losses due to reduced stands and costs associated with replanting. This study compared the response of these 40 SoyNAM parent genotypes to seed- and seedling-rot pathogens, Fusarium graminearum, Phytophthora sojae, Globisporangium ultimum var. ultimum, G. ultimum var. sporangiiferum, and G. irregulare groups 1, 2 and G. cryptoirregulare. None of the parental genotypes conferred high levels of resistance to F. graminearum nor G. ultimum var. sporangiiferum. Of the 40 parental genotypes, 15 were resistant to P. sojae OH1 (vir 7) indicating they contain Rps genes while the remaining (including the common parent IA 3023) do not have Rps genes. Based on inoculations with known P. sojae pathotypes, Rps1c was the most common followed by Rps1a as both Rps genes confer resistance to isolates OH4 (vir 1a, 1c, 7) and OH25 (vir 1a, 1b,1c, 1k, 7). Eight of the SoyNAM parents had higher levels of partial resistance to P. sojae than Conrad (cultivar with moderate resistance). There was moderate resistance to G. ultimum var ultimum among the 40 parents and to G. irregulare subpopulations among the 6 parents that were evaluated. The SoyNAM parental lines and populations are an excellent resource available for soybean breeders to advance the development of new cultivars with improved resistance to some soilborne pathogens.

Phytophthora root and stem rot is a major constraint to soybean [Glycine max (L.) Merr.] production worldwide. Deployment of single dominant Resistance to Phytophthora sojae (Rps) genes are an effective management strategy for this disease. However, due to increasing diversity in P. sojae populations for pathotype, new effective Rps genes are needed. Two recombinant inbred line (RIL) populations, each derived from a cross with Williams (susceptible) and resistant accessions PI 407974B and PI 424487B, were evaluated for resistance with one or more P. sojae pathotypes: OH1 (vir 7), OH4 (vir 1a, 1c, 7), OH7 (vir 1a, 3a, 3c, 4, 5, 6, 7), OH25 (vir 1a, 1b, 1c, 1k, 7), and 1.S.1.1 (1a, 1b, 1k, 2, 3a, 3c, 4, 5, 6, 7, 8). Molecular maps were assembled with BARCSoySNP6K BeadChip, simple-sequence repeat, and Kompetitive Allele Specific polymerase chain reaction markers. A total of three Rps loci were mapped, one near Rps1 on Chromosome 3 and two near Rps4/6 on chromosome 18. Quantitative trait loci and straight linkage maps confirmed the loci. Resistance to P. sojae pathotypes 1.S.1.1 and/or OH7 was mapped to Chromosome 3 in the PI 407974B RIL population. PI 407974B and PI 424487B RIL populations have Rps loci on chromosome 18 toward OH4 and OH25, respectively, which are near the Rps4/6 region. Although these PIs may have novel Rps genes/alleles and could assist in the deployment and pyramiding of resistance against P. sojae, care should be taken because these may condition defense reactions to some P. sojae pathotypes but not to all., Funding for this project was provided by the United Soybean Board, the Ohio Soybean Council, and the Ohio State University Center for Applied Plant Sciences, College of Food, Agricultural and Environmental Sciences, Center for Soybean Research. In addition, we wish to acknowledge support from the Molecular Cellular Imaging Center at Ohio Agricultural Research and Development Center. Salaries and research support for this project was provided by State and Federal Funds appropriated to the Ohio Agricultural Research and Development Center, The Ohio State University and the National Institute of Food and Agriculture, USDA, Hatch projects Development of Disease Management Strategies for Soybean Pathogens in Ohio OHO01303 and Genetic Analysis of Soybean Added-Value Traits and Soybean Variety Development for Ohio OHO01279.

Multiple species of Phytophthora, Phytopythium, and Pythium cause detrimental effects to soybean and more than one species have often been recovered from symptomatic soybean seedlings or roots, suggesting that these occur as species complexes. Our objective was to assess the species diversity and abundance of Phytophthora, Phytopythium, and Pythium spp. associated with soybean seedlings at two different incubation temperatures from soils collected from the same 81-ha farm but from five fields with a different history of production practices. To determine the effect of incubation temperature on the species detected, soils were baited with soybean seedlings at 15 and 25°C. At each temperature, isolates were directly recovered from seedling roots and the associated rhizosphere soil was assayed via amplicon sequencing with oomycete-specific primers. Regardless of incubation temperature or field site, the species Pythium sylvaticum and Pythium ultimum were isolated through the baiting procedure and these, along with Pythium acrogynum, Pythium attrantheridium, and Pythium heterothallicum, were detected with amplicon sequencing. Some Pythium spp. were more abundant at 15°C while others were favored at 25°C. This study demonstrated that the species diversity of Phytophthora, Phytopythium, and Pythium detected on seedling roots and in the rhizosphere of soybean can be influenced by incubation temperature. This may explain, in part, the differences in the species composition reported from previous surveys and suggests that production practices associated with each of the sampled fields may have influenced the species complexes of Phytophthora, Phytopythium, and Pythium.

The roles that specific cultivars with different levels and types of resistance may play on the diversity and abundance of Phytophthora, Phytopythium, and Pythium spp. communities were compared. Three soybean cultivars were planted in a total of 11 different high disease environments in Ohio and seedlings were collected for direct isolation and amplicon sequencing of the rhizosphere soil. Plant population and yields were lower for the moderately susceptible cultivar Sloan compared with the cultivars Kottman, with high partial resistance to Phytophthora sojae, and Lorain, with high partial resistance to both Pythium spp. and Phytophthora sojae. Based on both direct isolation and amplicon sequencing, distinct communities with key indicator species were identified at each environment, including differences at 14 and 25 day after planting for two environments. The environment had the largest influence on the community composition. Soybean genotype did influence the abundance of the hemibiotroph Phytophthora sojae but not species of Phytopythium or Pythium. In addition, from all environments and cultivars, operational taxonomic units (OTUs) classified as Pythium attrantheridium, P. heterothallicum, and P. sylvaticum were always detected. This is also the first report of an OTU of P. periilum from soils and the first one to recover undescribed species that match the sequence of Pythium sp. CAL2011f from soybean seedlings in Ohio This study highlights which species to target for the identification and development of resistant cultivars or for fungicide screening.

Soybean (Glycine max [L.] Merrill) yield losses as a result of plant diseases were estimated by university and government plant pathologists in 29 soybean producing states in the United States and in Ontario, Canada, from 2015 through 2019. In general, the estimated losses that resulted from each of 28 plant diseases or pathogens varied by state or province as well as year. Soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) caused more than twice as much loss as any other disease during the survey period. Seedling diseases (caused by various pathogens), Sclerotinia stem rot (white mold) (caused by Sclerotinia sclerotiorum [Lib.] de Bary), and sudden death syndrome (caused by Fusarium virguliforme O’Donnell & T. Aoki) caused the next greatest yield losses, in descending order. Following SCN, the most damaging diseases in the northern United States and Ontario differed from those in the southern United States. The estimated mean economic loss from all soybean diseases, averaged across the United States and Ontario, Canada was US$45 per acre (US$111 per hectare). The outcome from the current survey will provide pertinent information regarding the important soybean diseases and their overall severity in the soybean crop and help guide future research and Extension efforts on managing soybean diseases.

Auxin (indole-3-acetic acid, IAA) has been implicated as a susceptibility factor in both beneficial and pathogenic molecular plant−microbe interactions. Previous studies have identified a large number of auxin-related genes underlying quantitative disease resistance loci (QDRLs) for Phytophthora sojae. Thus, we hypothesized that auxin may be involved the P. sojae−soybean interaction. The levels of IAA and related metabolites were measured in mycelia and media supernatant as well as in mock and inoculated soybean roots in a time course assay. The expression of 11 soybean Pin-formed (GmPIN) auxin efflux transporter genes was also examined. Tryptophan, an auxin precursor, was detected in the P. sojae mycelia and media supernatant. During colonization of roots, levels of IAA and related metabolites were significantly higher in both moderately resistant Conrad and moderately susceptible Sloan inoculated roots compared with mock controls at 48 h postinoculation (hpi) in one experiment and at 72 hpi in a second, with Sloan accumulating higher levels of the auxin catabolite IAA-Ala than Conrad. Additionally, one GmPIN at 24 hpi, one at 48 hpi, and three at 72 hpi had higher expression in inoculated compared with the mock control roots in Conrad. The ability of resistant cultivars to cope with auxin accumulation may play an important role in quantitative disease resistance. Levels of jasmonic acid (JA), another plant hormone associated with defense responses, were also higher in inoculated roots at these same time points, suggesting that JA also plays a role during the later stages of infection.

Genomic prediction of quantitative resistance toward Phytophthora sojae indicated that genomic selection may increase breeding efficiency. Statistical model and marker set had minimal effect on genomic prediction with > 1000 markers. Quantitative disease resistance (QDR) toward Phytophthora sojae in soybean is a complex trait controlled by many small-effect loci throughout the genome. Along with the technical and rate-limiting challenges of phenotyping resistance to a root pathogen, the trait complexity can limit breeding efficiency. However, the application of genomic prediction to traits with complex genetic architecture, such as QDR toward P. sojae, is likely to improve breeding efficiency. We provide a novel example of genomic prediction by measuring QDR to P. sojae in two diverse panels of more than 450 plant introductions (PIs) that had previously been genotyped with the SoySNP50K chip. This research was completed in a collection of diverse germplasm and contributes to both an initial assessment of genomic prediction performance and characterization of the soybean germplasm collection. We tested six statistical models used for genomic prediction including Bayesian Ridge Regression; Bayesian LASSO; Bayes A, B, C; and reproducing kernel Hilbert spaces. We also tested how the number and distribution of SNPs included in genomic prediction altered predictive ability by varying the number of markers from less than 50 to more than 34,000 SNPs, including SNPs based on sequential sampling, random sampling, or selections from association analyses. Predictive ability was relatively independent of statistical model and marker distribution, with a diminishing return when more than 1000 SNPs were included in genomic prediction. This work estimated relative efficiency per breeding cycle between 0.57 and 0.83, which may improve the genetic gain for P. sojae QDR in soybean breeding programs.

Some Pythium spp. cause damping off and root rot in soybeans and other crop species. One of the most effective management tools to reduce disease is host resistance; however, little is known about resistance in soybean to Pythium spp. The soybean nested associated mapping (SoyNAM) parent lines are a set of germplasms that were crossed to a single hub parent to create recombinant inbred line populations for the purpose of mapping agronomic traits. The SoyNAM parents were screened for resistance to Pythium lutarium, Pythium oopapillum, Pythium sylvaticum, and Pythium torulosum in separate assays to evaluate seed and root rot severity. Of the 40 SoyNAM parents, only ‘Maverick’ was resistant to the four species tested; however, 13 were resistant to three species. Other lines were resistant to two, one, or none of the species tested. Correlations between seed and root rot severity for the lines assessed were weak or insignificant. Results indicate that mechanisms of resistance to seed and root rot caused by Pythium spp. may not necessarily be the same.

Phytophthora, Phytopythium, and Pythium species that cause early-season seed decay and pre-emergence and post-emergence damping off of soybean are most commonly managed with seed treatments. The phenylamide fungicides metalaxyl and mefenoxam, and ethaboxam are effective toward some but not all species. The primary objective of this study was to evaluate the efficacy of ethaboxam in fungicide mixtures and compare those with other fungicides as seed treatments to protect soybean against Pythium, Phytopythium, and Phytophthora species in both high-disease field environments and laboratory seed plate assays. The second objective was to evaluate these seed treatment mixtures on cultivars that have varying levels and combinations of resistance to these soilborne pathogens. Five of eight environments received adequate precipitation in the 14 days after planting for high levels of seedling disease development and treatment evaluations. Three environments had significantly greater stands, and three had significantly greater yield when ethaboxam was used in the seed treatment mixture compared with treatments containing metalaxyl or mefenoxam alone. Three fungicide formulations significantly reduced disease severity compared with nontreated in the seed plate assay for 17 species. However, the combination of ethaboxam plus metalaxyl in a mixture was more effective than either fungicide alone against some Pythium and Phytopythium species. Overall, our results indicate that the addition of ethaboxam to a fungicide seed treatment is effective in reducing seed rot caused by these pathogens commonly isolated from soybean in Ohio but that these effects can be masked when cultivars with resistance are planted.

Heterodera glycines, the soybean cyst nematode, and Macrophomina phaseolina, causal agent of charcoal rot, are economically important soybean pathogens. The impact and effect of these pathogens on soybean yield in coinfested fields in the Midwest production region is not known. Both pathogens are soilborne, with spatially aggregated distribution and effects. Spatial regression analysis, therefore, is an appropriate method to account for the spatial dependency in either the dependent variable or regression error term from data produced in fields naturally infested with H. glycines and M. phaseolina. The objectives of this study were twofold: to evaluate the combined effect of H. glycines and M. phaseolina on soybean yield in naturally infested commercial fields with ordinary least squares and spatial regression models; and to evaluate, under environmentally controlled conditions, the combined effect of H. glycines and M. phaseolina through nematode reproduction and plant tissue fungal colonization. Six trials were conducted in fields naturally infested with H. glycines and M. phaseolina in Ohio. Systematic-grid sampling was used to determine the population densities of H. glycines and M. phaseolina, and soybean yield estimates. Though not used in any statistical analysis, M. phaseolina colony forming units from plant tissue, charcoal rot severity, and H. glycines type were also recorded and summarized. In two greenhouse experiments, treatments consisted of H. glycines alone, M. phaseolina alone, and coinfestation of soybean with both pathogens. Moran’s I test indicated that the yield from five fields was spatially correlated (P < 0.05) and aggregated. In these fields, to account for spatial dependence, spatial regression models were fitted to the data. Spatial regression analyses revealed a significant interaction effect between H. glycines and M. phaseolina on soybean yield for fields with high initial population densities of both pathogens. In the greenhouse experiments, H. glycines reproduction was significantly (P < 0.05) reduced in the presence of M. phaseolina; however, soybean tissue fungal colonization was not affected by the presence of H. glycines. The direct mechanisms by which H. glycines and M. phaseolina interact were not demonstrated in this study. Future studies must be conducted in the field and greenhouse to better understand this interaction effect.

Accessory genes are variably present among members of a species and are a reservoir of adaptive functions. In bacteria, differences in gene distributions among individuals largely result from mobile elements that acquire and disperse accessory genes as cargo. In contrast, the impact of cargo-carrying elements on eukaryotic evolution remains largely unknown. Here, we show that variation in genome content within multiple fungal species is facilitated by Starships, a novel group of massive mobile elements that are 110 kb long on average, share conserved components, and carry diverse arrays of accessory genes. We identified hundreds of Starship-like regions across every major class of filamentous Ascomycetes, including 28 distinct Starships that range from 27-393 kb and last shared a common ancestor ca. 400 mya. Using new long-read assemblies of the plant pathogen Macrophomina phaseolina, we characterize 4 additional Starships whose past and ongoing activities contribute to standing variation in genome structure and content. One of these elements, Voyager, inserts into 5S rDNA and contains a candidate virulence factor whose increasing copy number has contrasting associations with pathogenic and saprophytic growth, suggesting Voyager’s activity underlies an ecological trade-off. We propose that Starships are eukaryotic analogs of bacterial integrative and conjugative elements based on parallels between their conserved components and may therefore represent the first known agents of active gene transfer in eukaryotes. Our results suggest that Starships have shaped the content and structure of fungal genomes for millions of years and reveal a new concerted route for evolution throughout an entire eukaryotic phylum.

Rhizoctonia solani AG-2-2IIIB is an important seedling pathogen of soybean in North America and other soybean-growing regions around the world. There is no information regarding the population genetics of field populations of R. solani associated with soybean seedling disease. More specifically, information regarding genetic diversity, the mode of reproduction, and the evolutionary factors that shape different R. solani populations separated in time and space are lacking. We exploited genotyping by sequencing as a tool to assess the genetic structure of R. solani AG-2-2IIIB populations from Illinois, Ohio, and Ontario and investigate the reproductive mode of this subgroup. Our results revealed differences in genotypic diversity among three populations, with the Ontario population having greatest diversity. An overrepresentation of multilocus genotypes (MLGs) and a rejection of the null hypothesis of random mating in all three populations suggested clonality within each population. However, phylogenetic analysis revealed long terminal multifurcating branches for most members of the Ontario population, suggesting a mixed reproductive mode for this population. Analysis of molecular variance revealed low levels of population differentiation, and sharing of similar MLGs among populations highlights the role of genotype flow as an evolutionary force shaping population structure of this subgroup.

In this study, four recombinant inbred line (RIL) soybean populations were screened for their response to infection by Pythium sylvaticum, Pythium irregulare, Pythium oopapillum, and Pythium torulosum. The parents, PI 424237A, PI 424237B, PI 408097, and PI 408029, had higher levels of resistance to these species in a preliminary screening and were crossed with “Williams,” a susceptible cultivar. A modified seed rot assay was used to evaluate RIL populations for their response to specific Pythium species selected for a particular population based on preliminary screenings. Over 2500 single-nucleotide polymorphism (SNP) markers were used to construct chromosomal maps to identify regions associated with resistance to Pythium species. Several minor and large effect quantitative disease resistance loci (QDRL) were identified including one large effect QDRL on chromosome 8 in the population of PI 408097 × Williams. It was identified by two different disease reaction traits in P. sylvaticum, P. irregulare, and P. torulosum. Another large effect QDRL was identified on chromosome 6 in the population of PI 408029 × Williams, and conferred resistance to P. sylvaticum and P. irregulare. These large effect QDRL will contribute toward the development of improved soybean cultivars with higher levels of resistance to these common soil-borne pathogens.

In this study, four recombinant inbred line (RIL) soybean populations were screened for their response to infection by Pythium sylvaticum, Pythium irregulare, Pythium oopapillum, and Pythium torulosum. The parents, PI 424237A, PI 424237B, PI 408097, and PI 408029, had higher levels of resistance to these species in a preliminary screening and were crossed with “Williams,” a susceptible cultivar. A modified seed rot assay was used to evaluate RIL populations for their response to specific Pythium species selected for a particular population based on preliminary screenings. Over 2500 single-nucleotide polymorphism (SNP) markers were used to construct chromosomal maps to identify regions associated with resistance to Pythium species. Several minor and large effect quantitative disease resistance loci (QDRL) were identified including one large effect QDRL on chromosome 8 in the population of PI 408097 × Williams. It was identified by two different disease reaction traits in P. sylvaticum, P. irregulare, and P. torulosum. Another large effect QDRL was identified on chromosome 6 in the population of PI 408029 × Williams, and conferred resistance to P. sylvaticum and P. irregulare. These large effect QDRL will contribute toward the development of improved soybean cultivars with higher levels of resistance to these common soil-borne pathogens.

Auxin (indole-3-acetic acid, IAA) has been implicated as a susceptibility factor in both beneficial and pathogenic molecular plant-microbe interactions. Previous studies have identified a large number of auxin-related genes underlying quantitative disease resistance loci (QDRLs) for

Phytophthora sojae causes Phytophthora root and stem rot of soybean and has been primarily managed through deployment of qualitative Resistance to P. sojae genes (Rps genes). The effectiveness of each individual or combination of Rps gene(s) depends on the diversity and pathotypes of the P. sojae populations present. Due to the complex nature of P. sojae populations, identification of more novel Rps genes is needed. In this study, phenotypic data from previous studies of 16 panels of plant introductions (PIs) were analyzed. Panels 1 and 2 consisted of 448 Glycine max and 520 G. soja, which had been evaluated for Rps gene response with a combination of P. sojae isolates. Panels 3 and 4 consisted of 429 and 460 G. max PIs, respectively, which had been evaluated using individual P. sojae isolates with complex virulence pathotypes. Finally, Panels 5–16 (376 G. max PIs) consisted of data deposited in the USDA Soybean Germplasm Collection from evaluations with 12 races of P. sojae. Using these panels, genome‐wide association (GWA) analyses were carried out by combining phenotypic and SoySNP50K genotypic data. GWA models identified two, two, six, and seven novel Rps loci with Panels 1, 2, 3, and 4, respectively. A total of 58 novel Rps loci were identified using Panels 5–16. Genetic and phenotypic dissection of these loci may lead to the characterization of novel Rps genes that can be effectively deployed in new soybean cultivars against diverse P. sojae populations.

Phytophthora sojae is one of the costliest soybean pathogens in the US. Quantitative disease resistance (QDR) is a vital part of Phytophthora disease management. In this study, QDR was measured in 478 and 495 plant introductions (PIs) towards P. sojae isolates OH.121 and C2.S1, respectively, in genome-wide association (GWA) analyses to identify genetic markers linked to QDR loci (QDRL). Populations were generated by sampling PIs from the US, the Republic of Korea, and the full collection of PIs maintained by the USDA. Additionally, a meta-analysis of QDRL reported from bi-parental studies was done to compare past and present findings. Twenty-four significant marker-trait associations were identified from the 478 PIs phenotyped with OH.121, and an additional 24 marker-trait associations were identified from the 495 PIs phenotyped with C2.S1. In total, 48 significant markers were distributed across 16 chromosomes and based on linkage analysis, represent a total of 44 QDRL. The majority of QDRL were identified with only one of the two isolates, and only a region on chromosome 13 was consistently identified. Regions on chromosomes 3, 13, and 17 were identified in previous GWA-analyses and were re-identified in this study. Five QDRL co-localized with P. sojae meta-QDRL identified from QDRL reported in previous biparental mapping studies. The remaining regions represent novel QDRL, in the soybean-P. sojae pathosystem and were primarily identified in germplasm from the Republic of Korea. Overall, the number of loci identified in this study highlights the complexity of QDR to P. sojae.

Two novel QTL for resistance to Pythium ultimum var. ultimum were identified in soybean using an Illumina SNP Chip and whole genome re-sequencing. Pythium ultimum var. ultimum is one of numerous Pythium spp. that causes severe pre- and post-emergence damping-off of seedlings and root rot of soybean [Glycine max (L.) Merr.]. The objective of this research was to identify quantitative trait loci (QTL) for resistance to P. ultimum var. ultimum in a recombinant inbred line population derived from a cross of 'Magellan' (moderately resistant) and PI 438489B (susceptible). Two different mapping approaches were utilized: the universal soybean linkage panel (USLP 1.0) and the bin map constructed from whole genome re-sequencing (WGRS) technology. Two genomic regions associated with variation in three disease-related parameters were detected using both approaches, with the bin map providing higher resolution. Using WGRS, the first QTL were mapped within a 350-kbp region on Chr. 6 and explained 7.5-13.5% of the phenotypic variance. The second QTL were positioned in a 260-kbp confidence interval on Chr. 8 and explained 6.3-16.8% of the phenotypic variation. Candidate genes potentially associated with disease resistance were proposed. High-resolution genetic linkage maps with a number of significant SNP markers could benefit marker-assisted breeding and dissection of the molecular mechanisms underlying soybean resistance to Pythium damping-off in 'Magellan.' Additionally, the outputs of this study may encourage more screening of diverse soybean germplasm and utilization of genome-wide association studies to understand the genetic basis of quantitative disease resistance.

Germplasm screening programs have primarily relied on inoculation with mycelia to determine the resistance reaction of soybean genotypes to Sclerotinia sclerotiorum. However, under field conditions, ascospores are the primary source of inoculum. Therefore, the objective of this study was to determine which components most accurately differentiate the resistance reaction of soybean genotypes inoculated with ascospores of S. sclerotiorum. Ascospores were produced in the laboratory and all of the experiments were carried out under controlled conditions with inoculations at flowering stage. Initially, inoculum densities of 1 × 104, 1 × 105 and 1 × 106 ascospores ml−1 were compared on six soybean genotypes with known resistance reactions. Disease symptoms developed on all genotypes and at all inoculum densities. The highest ascospore concentration increased infection efficiency but it was not correlated with an increase in lesion length. Components of resistance were then measured on a set of 17 cultivars with known resistance reactions at 1 × 105 ascospores ml−1. Resistance reactions could be differentiated based on the level of infection efficiency and lesion length on the main stem. Although inoculation with ascospores presents some limitations such as the time required for inoculum production as well as the time and space required for plant growth, it has the potential to be used to complement other methods for the characterization of resistance of soybean genotypes.

Phytophthora sojae Kaufmann & Gerdemann has been a yield limiting factor for more than 60 years on soybean (Glycine max L.) in Ohio and other regions where soils are poorly drained. Soybean is the ...

Isolates of Cercospora sojina, causal agent of frogeye leaf spot of soybean (Glycine max), were collected across Alabama, Arkansas, Delaware, Illinois, Indiana, Iowa, Kentucky, Louisiana, Mississippi, Missouri, North Carolina, Ohio, Tennessee, and Virginia and were evaluated for quinone outside inhibitor (QoI) fungicide resistance. Collection of these isolates from these 14 states occurred between 2010 and 2017. QoI fungicide-resistant C. sojina isolates were detected in all 14 states surveyed and represent a total of 240 counties or parishes. In 2017, these 240 counties and parishes represented approximately 13% of the harvested soybean hectares in the United States. In light of this widespread occurrence of QoI fungicide-resistant C. sojina isolates, management of frogeye leaf spot should focus on integrated management practices such as planting resistant soybean cultivars, rotating with nonhost crops, and tilling to speed up decomposition of infested soybean residue. When foliar fungicide application is warranted, fungicide products that contain active ingredients from chemistry classes other than the QoI class should be applied for frogeye leaf spot management.

Recent outbreaks of Sclerotinia stem rot (SSR) of soybean in Ohio, along with new fungicides and cultivars with resistance to this disease, have led to a renewed interest in studies to update disease management guidelines. The effect of host resistance (in moderately resistant [MR] and moderately susceptible [MS] cultivars) and chemical control on SSR and yield was evaluated in 12 environments from 2014 to 2016. The chemical treatments evaluated were an untreated check, four fungicides (boscalid, picoxystrobin, pyraclostrobin, and thiophanate-methyl), and one herbicide (lactofen) applied at soybean growth stage R1 (early flowering) alone or at R1 followed by a second application at R2 (full flowering). SSR developed in 6 of 12 environments, with mean disease incidence in the untreated check of 2.5 to 41%. The three environments with high levels of SSR (disease incidence in the untreated check >20%) were used for further statistical analysis. There were significant effects (P < 0.05) of soybean cultivar and chemical treatment on SSR levels. Significantly lower levels of SSR were observed in MR cultivars. Both boscalid and lactofen reduced SSR but did not increase yield. Pyraclostrobin increased SSR compared with the untreated check in the three environments with high levels of disease. In the six fields where SSR did not develop, chemical treatment did not increase yield, nor was the yield from the MR cultivar significantly different from the MS cultivar. For Ohio, MR cultivars alone were effective for management of SSR in soybean fields where this disease has historically occurred.

A major novel quantitative disease resistance locus, qRfg_Gm06, for Fusarium graminearum was genetically mapped to chromosome 6. Genomic-assisted haplotype analysis within this region identified three putative candidate genes. Fusarium graminearum causes seed, root rot, and seedling damping-off in soybean which contributes to reduced stands and yield. A cultivar Magellan and PI 567516C were identified with low and high levels of partial resistance to F. graminearum, respectively. Quantitative disease resistance loci (QDRL) were mapped with 241 F7:8 recombinant inbred lines (RILs) derived from a cross of Magellan × PI 567516C. Phenotypic evaluation for resistance to F. graminearum used the rolled towel assay in a randomized incomplete block design. The genetic map was constructed from 927 polymorphic single nucleotide polymorphism (SNP) and simple sequence repeat (SSR) markers. One major QDRL qRfg_Gm06 was detected and mapped to chromosome 6 with a LOD score of 20.3 explaining 40.2% of the total phenotypic variation. This QDRL was mapped to a ~400 kb genomic region of the Williams 82 reference genome. Genome mining of this region identified 14 putative candidate disease resistance genes. Haplotype analysis of this locus using whole genome re-sequencing (WGRS) of 106 diverse soybean lines narrowed the list to three genes. A SNP genotyping Kompetitive allele-specific PCR (KASP) assay was designed for one of the genes and was validated in a subset of the RILs and all 106 diverse lines.

Annual decreases in soybean (Glycine max L. Merrill) yield caused by diseases were estimated by surveying university-affiliated plant pathologists in 28 soybean-producing states in the United States and in Ontario, Canada, from 2010 through 2014. Estimated yield losses from each disease varied greatly by state or province and year. Over the duration of this survey, soybean cyst nematode (SCN) (Heterodera glycines Ichinohe) was estimated to have caused more than twice as much yield loss than any other disease. Seedling diseases (caused by various pathogens), charcoal rot (caused by Macrophomina phaseolina (Tassi) Goid), and sudden death syndrome (SDS) (caused by Fusarium virguliforme O’Donnell & T. Aoki) caused the next greatest estimated yield losses, in descending order. The estimated mean economic loss due to all soybean diseases, averaged across U.S. states and Ontario from 2010 to 2014, was $60.66 USD per acre. Results from this survey will provide scientists, breeders, governments, and educators with soybean yield-loss estimates to help inform and prioritize research, policy, and educational efforts in soybean pathology and disease management.

Greenhouse and laboratory screening methods for assessing potato foliage for resistance to late blight were compared using 15 cultivars and advanced breeding selections with known field response to late blight. Screening methods included greenhouse inoculation of plants in several age classes, and laboratory assays of detached leaflets, leaf disks, and stem cuttings. Greenhouse inoculation of plants 7 to 11 weeks after planting, near the time of flowering, corresponded best to results obtained in field evaluations, but there were significant differences in disease severity between separate greenhouse tests. This is consistent with variation in late blight severity on a year-to-year basis when cultivars are compared in the field. The greenhouse inoculation method allowed for testing of several components of partial resistance, such as infection efficiency and lesion growth rate, which may exist for each cultivar. Laboratory assays proved less reliable than greenhouse assays for overall ratings of partial resistance, but could be useful for measuring specific components of resistance. Screening evaluations for late blight resistance should include standard cultivars with known reaction to Phytophthora infestans to reference the disease potential within the screening evaluation.

Dorrance, A. E., Inglis, D. A., Derie, M. L., Brown, C. R., Goodwin, S. B., Fry, W. E., and Deahl, K. L. 1999. Characterization of Phytophthora infestans populations in western Washington. Plant Dis. 83:423-428. The first detection in the United States of isolates of Phytophthora infestans having metalaxyl insensitivity and complex pathotypes occurred in western Washington during the early 1990s. To determine the genetic structure of the current population in western Washington, a total of 115 isolates of P. infestans were obtained during 1996 from infected tubers or foliage of potato, tomato, nightshade, and bittersweet thr oughout the region. An additional 45 isolates were collected from a single field. Based on mating type, metalaxyl-insensitivity, and molecular markers (allozymes of glucose-6-phosphate isomerase, peptidase, and RG57 DNA fingerprint), all of the isolates were A1 mating type and had the US-11 multilocus genotype. Analyses of an additional 120 isolates collected during 1997 from potato, tomato, and nightshade were performed. As in 1996, US-11 was the predominant genotype detected on all three hosts. However, three additional A2 mating type genotypes were also detected: US-7, US-8, and US-14. These three genotypes represent the first A2 mating type isolates detected in western Washington. Most of a subset of 60 isolates infected 4 to 7 of the 10 potato differentials tested. This included 90% of the isolates collected in 1996 (all US-11), plus 72% of the US-11 and 100% of the US-8 and US14 isolates collected during 1997. Virulence phenotypes in this region are complex even without the selection pressure of R-genes in the local commercial cultivars. The apparent increase in genetic variation observed in populations of P. infestans in western Washington from 1996 to 1997 most likely occurred by migration rather than by sexual recombination.