Detection and discovery of plant viruses in soybean by metagenomic sequencing.

BACKGROUND: Viruses negatively impact soybean production by causing diseases that affect yield and seed quality. Newly emerging or re-emerging viruses can also threaten soybean production because current control measures may not be effective against them. Furthermore, detection and characterization of new plant viruses requires major efforts when no sequence or antibody-based resources are available. METHODS: In this study, soybean fields were scouted for virus-like disease symptoms during the 2016-2019 growing seasons. Total RNA was extracted from symptomatic soybean parts, cDNA libraries were prepared, and RNA sequencing was performed using high-throughput sequencing (HTS). A custom bioinformatic workflow was used to identify and assemble known and unknown virus genomes. RESULTS: Several viruses were identified in single or mixed infections. Full- or nearly full-length genomes were generated for tobacco streak virus (TSV), alfalfa mosaic virus (AMV), tobacco ringspot virus (TRSV), soybean dwarf virus (SbDV), bean pod mottle virus (BPMV), soybean vein necrosis virus (SVNV), clover yellow vein virus (ClYVV), and a novel virus named soybean ilarvirus 1 (SIlV1). Two distinct ClYVV isolates were recovered, and their biological properties were investigated in Nicotiana benthamiana, broad bean, and soybean. In addition to infections by individual viruses, we also found that mixed viral infections in various combinations were quite common. CONCLUSIONS: Taken together, the results of this study showed that HTS-based technology is a valuable diagnostic tool for the identification of several viruses in field-grown soybean and can provide rapid information about expected viruses as well as viruses that were previously not detected in soybean.

Influence of Fusarium virguliforme Temporal Colonization of Corn, Tillage, and Residue Management on Soybean Sudden Death Syndrome and Soybean Yield.

The asymptomatic host range of Fusarium virguliforme includes corn, a common crop rotated with soybean that we hypothesize may alter F. virguliforme population dynamics and disease management. A field-based approach explored the temporal dynamics of F. virguliforme colonization of corn and soybean roots under different tillage and residue managements. Experiments were conducted in Iowa, Indiana, Michigan, and Wisconsin, United States and Ontario, Canada from 2016 to 2018. Corn and soybean roots were sampled at consecutive timepoints between 1 and 16 weeks after planting. DNA was extracted from all roots and analyzed by real-time quantitative PCR for F. virguliforme quantification. Trials were rotated between corn and soybean, containing a two-by-two factorial of tillage (no-tilled or tilled) and corn residue (with or without) in several experimental designs. In 2016, low amounts (approximately 100 fg per 10 mg of root tissue) of F. virguliforme were detected in the inoculated Iowa, Indiana, and Michigan locations and noninoculated Wisconsin corn fields. However, in 2017, greater levels of F. virguliforme DNA were detected in Iowa, Indiana, and Michigan across sampling timepoints. Tillage practices showed inconsistent effects on F. virguliforme root colonization and sudden death syndrome (SDS) foliar symptoms among trials and locations. However, residue management did not alter root colonization of corn or soybean by F. virguliforme. Plots with corn residue had greater SDS foliar disease index in Iowa in 2016. However, this trend was not observed across the site-years, indicating that corn residue may occasionally increase SDS foliar symptoms depending on the disease level and soil and weather factors.

Meta-Analysis of Soybean Yield Response to Foliar Fungicides Evaluated from 2005 to 2018 in the United States and Canada.

Random-effect meta-analyses were performed on data from 240 field trials conducted between 2005 and 2018 across nine U.S. states and Ontario, Canada, to quantify the yield response of soybean after application of foliar fungicides at beginning pod (R3) stage. Meta-analysis showed that the overall mean yield response when fungicide was used compared with not applying a fungicide was 2.7% (110 kg/ha). Moderator variables were also investigated and included fungicide group, growing season, planting date, and base yield, which all significantly influenced the yield response. There was also evidence that precipitation from the time of planting to the R3 growth stage influenced yield when fungicide was used (P = 0.059). Fungicides containing a premix of active ingredients from multiple groups (either two or three ingredients) increased the yield by 3.0% over not applying a fungicide. The highest and lowest yield responses were observed in 2005 and 2007, respectively. Better yield response to fungicides (a 3.0% increase) occurred when soybean crops were planted not later than 21 May and when total precipitation between planting and the R3 application date was above historic averages. Temperatures during the season did not influence the yield response. Yield response to fungicide was higher (a 4.7% increase) in average yield category (no spray control yield 2,878 to 3,758 kg/ha) and then gradually decreased with increasing base yield. Partial economic analyses indicated that use of foliar fungicides is less likely to be profitable when foliar diseases are absent or at low levels.

Relationship Between Sudden Death Syndrome caused by Fusarium virguliforme and Soybean Yield: A Meta-Analysis.

In total, 52 uniform field experiments were conducted in Illinois, Indiana, Iowa, Michigan, South Dakota, and Wisconsin in the United States and Ontario, Canada from 2013 to 2017 comparing crop protection products against sudden death syndrome (SDS) of soybean. Data were analyzed using meta-analytic models to summarize the relationship between foliar disease index (FDX) and yield. For each study, correlation and regression analyses were performed separately to determine three effect sizes: Fisher's transformation of correlation coefficients (Z r ), intercept (ß0), and slope (ß1). Random- and mixed-effect meta-analyses were used to summarize the effect sizes. Study- and location-specific moderator variables FDX (low < 10% and high ≥ 10%), date of planting (early = prior to 7 May, conventional = 7 to 21 May, and late = after 21 May) cultivar (susceptible and partially resistant to SDS), study location, and growing season were used as fixed effects. The overall mean effect sizes of transformed correlation coefficient [Formula: see text] r was -0.41 and different from zero (P < 0.001), indicating that yield was negatively correlated with FDX. The [Formula: see text] r was affected by disease level (P < 0.01) and cultivar (P = 0.02), with a greater effect at higher disease levels and with susceptible cultivars. The mean [Formula: see text] 0 was 4,121 kg/ha and mean [Formula: see text] 1 was -21 kg/ha/% FDX and were different from zero (P < 0.01). Results from these data indicate that, for every unit of FDX increase, yield was decreased by 0.5%. Study locations and year affected the [Formula: see text] 0 , whereas none of the moderator variables significantly affected [Formula: see text] 1.

Effect of Seed Treatment and Foliar Crop Protection Products on Sudden Death Syndrome and Yield of Soybean.

Sudden death syndrome (SDS), caused by Fusarium virguliforme, is an important soilborne disease of soybean. Risk of SDS increases when cool and wet conditions occur soon after planting. Recently, multiple seed treatment and foliar products have been registered and advertised for management of SDS but not all have been tested side by side in the same field experiment at multiple field locations. In 2015 and 2016, seed treatment fungicides fluopyram and thiabendazole; seed treatment biochemical pesticides citric acid and saponins extract of Chenopodium quinoa; foliar fungicides fluoxastrobin + flutriafol; and an herbicide, lactofen, were evaluated in Illinois, Indiana, Iowa, Michigan, South Dakota, Wisconsin, and Ontario for SDS management. Treatments were tested on SDS-resistant and -susceptible cultivars at each location. Overall, fluopyram provided the highest level of control of root rot and foliar symptoms of SDS among all the treatments. Foliar application of lactofen reduced foliar symptoms in some cases but produced the lowest yield. In 2015, fluopyram reduced the foliar disease index (FDX) by over 50% in both resistant and susceptible cultivars and provided 8.9% yield benefit in susceptible cultivars and 3.5% yield benefit in resistant cultivars compared with the base seed treatment (control). In 2016, fluopyram reduced FDX in both cultivars by over 40% compared with the base seed treatment. For yield in 2016, treatment effect was not significant in the susceptible cultivar while, in the resistant cultivar, fluopyram provided 3.5% greater yield than the base seed treatment. In this study, planting resistant cultivars and using fluopyram seed treatment were the most effective tools for SDS management. However, plant resistance provided an overall better yield-advantage than using fluopyram seed treatment alone. Effective seed treatments can be an economically viable consideration to complement resistant cultivars for managing SDS.

Inoculation Method Impacts Symptom Development Associated with Diaporthe aspalathi, D. caulivora, and D. longicolla on Soybean (Glycine max).

One hundred fifty-two Diaporthe isolates were recovered from symptomatic soybean (Glycine max) stems sampled from the U.S. states of Iowa, Indiana, Kentucky, Michigan, and South Dakota. Using morphology and DNA sequencing, isolates were identified as D. aspalathi (8.6%), D. caulivora (24.3%), and D. longicolla (67.1%). Aggressiveness of five isolates each of the three pathogens was studied on cultivars Hawkeye (D. caulivora and D. longicolla) and Bragg (D. aspalathi) using toothpick, stem-wound, mycelium contact, and spore injection inoculation methods in the greenhouse. For D. aspalathi, methods significantly affected disease severity (P < 0.001) and pathogen recovery (P < 0.001). The relative treatment effects (RTE) of stem-wound and toothpick methods were significantly greater than for the other methods. For D. caulivora and D. longicolla, a significant isolate × method interaction affected disease severity (P < 0.05) and pathogen recovery (P < 0.001). Significant differences in RTEs were observed among D. caulivora and D. longicolla isolates only when the stem-wound and toothpick methods were used. Our study has determined that the stem-wound and toothpick methods are reliable to evaluate the three pathogens; however, the significant isolate × method interactions for D. caulivora and D. longicolla indicate that multiple isolates should also be considered for future pathogenicity studies.

Effects of Pre- and Postanthesis Applications of Demethylation Inhibitor Fungicides on Fusarium Head Blight and Deoxynivalenol in Spring and Winter Wheat.

Anthesis is generally recommended as the optimum growth stage for applying a foliar fungicide to manage Fusarium head blight (FHB) and the Fusarium-associated toxin deoxynivalenol (DON) in wheat. However, because it is not always possible to treat fields at anthesis, studies were conducted to evaluate pre- and postanthesis treatment options for managing FHB and DON in spring and winter wheat. Network meta-analytical models were fitted to data from 19 years of fungicide trials, and log response ratio ([Formula: see text]) and approximate percent control ([Formula: see text]) relative to a nontreated check were estimated as measures of the effects of six treatments on FHB index (IND: mean percentage of diseased spikelets per spike) and DON. The evaluated treatments consisted of either Caramba (metconazole) applied early (at heading [CE]), at anthesis (CA), or late (5 to 7 days after anthesis; CL), or Prosaro (prothioconazole + tebuconazole) applied at the same three times and referred to as PE, PA, and PL, respectively. All treatments reduced mean IND and DON relative to the nontreated check, but the magnitude of the effect varied with timing and wheat type. CA and PA resulted in the highest [Formula: see text] values for IND, 52.2 and 51.5%, respectively, compared with 45.9% for CL, 41.3% for PL, and less than 33% for CE and PE. Anthesis and postanthesis treatments reduced mean IND by 14.9 to 29.7% relative to preanthesis treatments. The estimated effect size was also statistically significant for comparisons between CA and CL and PA and PL; CA reduced IND by 11.7% relative to CL, whereas PA reduced the disease by 17.4% relative to PL. Differences in efficacy against IND between pairs of prothioconazole + tebuconazole and metconazole treatments applied at the same timing (CE versus PE, CA versus PA, and CL versus PL) were not statistically significant. However, CA and CL outperformed PA and PL by 7 and 12.8%, respectively, in terms of efficacy against DON. All application programs had comparable efficacy against IND between spring and winter wheat types, but efficacy against DON was 10 to 16% greater for spring than winter wheat for applications made at or after anthesis. All programs led to an increase in mean grain yield and test weight relative to the nontreated check.

Benefits and Profitability of Fluopyram-Amended Seed Treatments for Suppressing Sudden Death Syndrome and Protecting Soybean Yield: A Meta-Analysis.

A meta-analytic approach was used to summarize data on the effects of fluopyram-amended seed treatment on sudden death syndrome (SDS) and yield of soybean (Glycine max L.) in over 200 field trials conducted in 12 U.S. states and Ontario, Canada from 2013 to 2015. In those trials, two treatments-the commercial base (CB), and CB plus fluopyram (CBF)-were tested, and all disease and yield data were combined to conduct a random-effects and mixed-effects meta-analysis (test of moderators) to estimate percent control and yield response relative to CB. Overall, a 35% reduction in foliar disease and 295 kg/ha (7.6%) increase in yield were estimated for CBF relative to CB. Sowing date and geographic region affected both estimates. The variation in yield response was explained partially by disease severity (19%), geographic region (8%), and sowing date (10%) but not by the resistance level of the cultivar. The probability of not offsetting the cost of fluopyram was estimated on a range of grain prices and treatment cost combinations. There was a high probability (>80%) of yield gains when disease level was high in any cost-price combinations tested but very low when the foliar symptoms of the disease were absent.

Oomycete Species Associated with Soybean Seedlings in North America-Part II: Diversity and Ecology in Relation to Environmental and Edaphic Factors.

Soybean (Glycine max (L.) Merr.) is produced across a vast swath of North America, with the greatest concentration in the Midwest. Root rot diseases and damping-off are a major concern for production, and the primary causal agents include oomycetes and fungi. In this study, we focused on examination of oomycete species distribution in this soybean production system and how environmental and soil (edaphic) factors correlate with oomycete community composition at early plant growth stages. Using a culture-based approach, 3,418 oomycete isolates were collected from 11 major soybean-producing states and most were identified to genus and species using the internal transcribed spacer region of the ribosomal DNA. Pythium was the predominant genus isolated and investigated in this study. An ecology approach was taken to understand the diversity and distribution of oomycete species across geographical locations of soybean production. Metadata associated with field sample locations were collected using geographical information systems. Operational taxonomic units (OTU) were used in this study to investigate diversity by location, with OTU being defined as isolate sequences with 97% identity to one another. The mean number of OTU ranged from 2.5 to 14 per field at the state level. Most OTU in this study, classified as Pythium clades, were present in each field in every state; however, major differences were observed in the relative abundance of each clade, which resulted in clustering of states in close proximity. Because there was similar community composition (presence or absence) but differences in OTU abundance by state, the ordination analysis did not show strong patterns of aggregation. Incorporation of 37 environmental and edaphic factors using vector-fitting and Mantel tests identified 15 factors that correlate with the community composition in this survey. Further investigation using redundancy analysis identified latitude, longitude, precipitation, and temperature as factors that contribute to the variability observed in community composition. Soil parameters such as clay content and electrical conductivity also affected distribution of oomycete species. The present study suggests that oomycete species composition across geographical locations of soybean production is affected by a combination of environmental and edaphic conditions. This knowledge provides the basis to understand the ecology and distribution of oomycete species, especially those able to cause diseases in soybean, providing cues to develop management strategies.

Oomycete Species Associated with Soybean Seedlings in North America-Part I: Identification and Pathogenicity Characterization.

Oomycete pathogens are commonly associated with soybean root rot and have been estimated to reduce soybean yields in the United States by 1.5 million tons on an annual basis. Limited information exists regarding the frequency and diversity of oomycete species across the major soybean-producing regions in North America. A survey was conducted across 11 major soybean-producing states in the United States and the province of Ontario, Canada. In 2011, 2,378 oomycete cultures were isolated from soybean seedling roots on a semiselective medium (CMA-PARPB) and were identified by sequencing of the internal transcribed spacer region of rDNA. Sequence results distinguished a total of 51 Pythium spp., three Phytophthora spp., three Phytopythium spp., and one Aphanomyces sp. in 2011, with Pythium sylvaticum (16%) and P. oopapillum (13%) being the most prevalent. In 2012, the survey was repeated, but, due to drought conditions across the sampling area, fewer total isolates (n = 1,038) were collected. Additionally, in 2012, a second semiselective medium (V8-RPBH) was included, which increased the Phytophthora spp. isolated from 0.7 to 7% of the total isolates. In 2012, 54 Pythium spp., seven Phytophthora spp., six Phytopythium spp., and one Pythiogeton sp. were recovered, with P. sylvaticum (14%) and P. heterothallicum (12%) being recovered most frequently. Pathogenicity and virulence were evaluated with representative isolates of each of the 84 species on soybean cv. Sloan. A seed-rot assay identified 13 and 11 pathogenic species, respectively, at 13 and 20°C. A seedling-root assay conducted at 20°C identified 43 species as pathogenic, having a significantly detrimental effect on the seedling roots as compared with the noninoculated control. A total of 15 species were pathogenic in both the seed and seedling assays. This study provides a comprehensive characterization of oomycete species present in soybean seedling roots in the major production areas in the United States and Ontario, Canada and provides a basis for disease management and breeding programs.

Genetic Diversity of Stenocarpella maydis in the Major Corn Production Areas of the United States.

The fungus Stenocarpella maydis (Berk.) B. Sutton, causal agent of Diplodia ear rot, is a prevalent corn (Zea mays L.) pathogen in the United States. Although S. maydis reduces grain quality, causes yield loss, and can produce mycotoxins in some countries, few studies have examined its biology and genetic diversity. We analyzed the genetic diversity of 174 S. maydis isolates sampled across the major corn production areas in the United States using nine different microsatellites. In all, 55 unique multilocus genotypes (MLG) were observed out of the 174 S. maydis isolates tested. After conducting a Bayesian clustering analysis by STRUCTURE, it was observed that the most probable number of genetic groups was two; however, no separation by their geographical location was identified. According to the minimum spanning network, the S. maydis population is linked across geographic regions of the United States but also contains private genotypes. Temporal diversity in the inoculum source was also observed at one location across 4 years. The haploid stage of S. maydis was confirmed and both mating type genes were amplified among selected isolates with unique MLG. We theorize that, although S. maydis is primarily an asexual fungus, sporadic cryptic recombination may occur, which could contribute to the genetic diversity observed in this study.

Effect of Soybean Cyst Nematode Resistance Source and Seed Treatment on Population Densities of Heterodera glycines, Sudden Death Syndrome, and Yield of Soybean.

A three-year study was conducted in Illinois, Indiana, Iowa, Michigan, and Ontario, Canada, from 2013 through 2015 to determine the effect of soybean (Glycine max) cultivars' source of soybean cyst nematode (SCN; Heterodera glycines) resistance on SCN population densities, sudden death syndrome (SDS; caused by Fusarium virguliforme), and yield of soybean. Five cultivars were evaluated with and without fluopyram seed treatment at each location. Cultivars with no SCN resistance had greater SDS severity, greater postharvest SCN egg counts (Pf), and lower yield than cultivars with plant introduction (PI) 548402 (Peking) and PI 88788-type of SCN resistance (P < 0.05). Cultivars with Peking-type resistance had lower Pf than those with PI 888788-type and no SCN resistance. In two locations with HG type 1.2-, cultivars with Peking-type resistance had greater foliar disease index (FDX) than cultivars with PI 88788-type. Fluopyram seed treatment reduced SDS and improved yield compared with a base seed treatment but did not affect SCN reproduction and Pf (P > 0.05). FDX and Pf were positively correlated in all three years (P < 0.01). Our results indicate that SDS severity may be influenced by SCN population density and HG type, which are important to consider when selecting cultivars for SCN management.

Fungicide and Cultivar Effects on Sudden Death Syndrome and Yield of Soybean.

The effect of fungicides on severity of sudden death syndrome (SDS; caused by Fusarium virguliforme), plant establishment, and soybean yield was evaluated in 12 field experiments conducted in Illinois, Indiana, Iowa, Michigan, and Ontario in 2013 and 2014. Two soybean cultivars that differed in susceptibility to SDS were planted in fields with a history of SDS or with artificial augmentation of F. virguliforme. Efficacy of seed, in-furrow, and foliar-applied fungicides was assessed. SDS levels varied across locations and years. Fluopyram applied on the seed or in-furrow reduced foliar disease index maximum up to 95% in 5 of the 12 experiments. In three experiments with significant (P < 0.10) treatment effect, fluopyram seed treatment improved yields up to 11% compared with the base seed treatment comprising prothioconazole + penflufen + metalaxyl and clothianidin + Bacillus firmus. Meta-analysis also indicated that the fluopyram seed treatment and in-furrow application were effective at reducing SDS and increasing yield relative to the control; however, the baseline disease influenced the yield and disease response to fungicide treatments. Treatment effect was not significant when disease pressure was low. The concentration of F. virguliforme DNA in soybean roots, measured by a specific real-time quantitative polymerase chain reaction assay, was not different among fungicide treatments in 9 of 10 experiments. Moderately resistant cultivars had less disease than susceptible cultivars, indicating that resistant cultivars in combination with fluopyram seed treatment or in-furrow application could provide effective management of SDS.

Effect of Planting Date, Seed Treatment, and Cultivar on Plant Population, Sudden Death Syndrome, and Yield of Soybean.

A 2-year study was conducted in Illinois, Indiana, Iowa, and Ontario in 2013 and 2014 to determine the effects of planting date, seed treatment, and cultivar on plant population, sudden death syndrome (SDS) caused by Fusarium virguliforme, and grain yield of soybean (Glycine max). Soybean crops were planted from late April to mid-June at approximately 15-day intervals, for a total of three to four plantings per experiment. For each planting date, two cultivars differing in SDS susceptibility were planted with and without fluopyram seed treatment. Mid-May plantings resulted in higher disease index compared with other planting dates in two experiments, early June plantings in three, and the remaining six experiments were not affected by planting date. Soil temperature at planting was not linked to SDS development. Root rot was greater in May plantings for most experiments. Resistant cultivars had significantly lower disease index than the susceptible cultivar in 54.5% of the experiments. Fluopyram reduced disease severity and protected against yield reductions caused by SDS in nearly all plantings and cultivars, with a maximum yield response of 1,142 kg/ha. Plant population was reduced by fluopyram seed treatment and early plantings in some experiments; however, grain yield was not affected by these reductions. Yields of plots planted in mid-June were up to 29.8% less than yields of plots planted in early May. The lack of correlation between early planting date and SDS severity observed in this study indicates that farmers do not have to delay planting in the Midwest to prevent yield loss due to SDS; cultivar selection combined with fluopyram seed treatment can reduce SDS in early-planted soybean (late April to mid May).

Effect of Glyphosate Application on Sudden Death Syndrome of Glyphosate-Resistant Soybean Under Field Conditions.

Sudden death syndrome (SDS), caused by Fusarium virguliforme, is an important yield limiting disease of soybean. Glyphosate is used to control weeds in soybean; however, its effect on SDS is not clearly understood. The objective of this study was to examine the impact of glyphosate on SDS, yield, and plant nutrition under field conditions. Fourteen field experiments were conducted in Iowa, Illinois, Indiana, Michigan, Wisconsin, and Ontario, Canada during 2011 to 2013. The experiment consisted of six treatment combinations of glyphosate and herbicides not containing glyphosate. Disease index was significantly different across the location-years, ranging from 0 to 65. The highest disease was noted in locations with irrigation, indicating that high soil moisture favors development of SDS. There were no effects of herbicide treatments or interactions on disease. The foliar disease index among the treatments over all years ranged from 9 to 13. Glyphosate-treatments also tended to yield more than treatments of herbicides not containing glyphosate. There were no interactions between glyphosate-treatments and total manganese in plant tissue. The interaction of glyphosate with other nutrients in plant tissue was inconclusive. This 14 location-year study demonstrated that glyphosate application did not increase SDS severity or adversely affect soybean yield under field conditions.

Population genomic analysis reveals geographic structure and climatic diversification for Macrophomina phaseolina isolated from soybean and dry bean across the United States, Puerto Rico, and Colombia.

Macrophomina phaseolina causes charcoal rot, which can significantly reduce yield and seed quality of soybean and dry bean resulting from primarily environmental stressors. Although charcoal rot has been recognized as a warm climate-driven disease of increasing concern under global climate change, knowledge regarding population genetics and climatic variables contributing to the genetic diversity of M. phaseolina is limited. This study conducted genome sequencing for 95 M. phaseolina isolates from soybean and dry bean across the continental United States, Puerto Rico, and Colombia. Inference on the population structure using 76,981 single nucleotide polymorphisms (SNPs) revealed that the isolates exhibited a discrete genetic clustering at the continental level and a continuous genetic differentiation regionally. A majority of isolates from the United States (96%) grouped in a clade with a predominantly clonal genetic structure, while 88% of Puerto Rican and Colombian isolates from dry bean were assigned to a separate clade with higher genetic diversity. A redundancy analysis (RDA) was used to estimate the contributions of climate and spatial structure to genomic variation (11,421 unlinked SNPs). Climate significantly contributed to genomic variation at a continental level with temperature seasonality explaining the most variation while precipitation of warmest quarter explaining the most when spatial structure was accounted for. The loci significantly associated with multivariate climate were found closely to the genes related to fungal stress responses, including transmembrane transport, glycoside hydrolase activity and a heat-shock protein, which may mediate climatic adaptation for M. phaseolina. On the contrary, limited genome-wide differentiation among populations by hosts was observed. These findings highlight the importance of population genetics and identify candidate genes of M. phaseolina that can be used to elucidate the molecular mechanisms that underly climatic adaptation to the changing climate.

Asymmetrical lineage introgression and recombination in populations of Aspergillus flavus: Implications for biological control.

Aspergillus flavus is an agriculturally important fungus that causes ear rot of maize and produces aflatoxins, of which B1 is the most carcinogenic naturally-produced compound. In the US, the management of aflatoxins includes the deployment of biological control agents that comprise two nonaflatoxigenic A. flavus strains, either Afla-Guard (member of lineage IB) or AF36 (lineage IC). We used genotyping-by-sequencing to examine the influence of both biocontrol agents on native populations of A. flavus in cornfields in Texas, North Carolina, Arkansas, and Indiana. This study examined up to 27,529 single-nucleotide polymorphisms (SNPs) in a total of 815 A. flavus isolates, and 353 genome-wide haplotypes sampled before biocontrol application, three months after biocontrol application, and up to three years after initial application. Here, we report that the two distinct A. flavus evolutionary lineages IB and IC differ significantly in their frequency distributions across states. We provide evidence of increased unidirectional gene flow from lineage IB into IC, inferred to be due to the applied Afla-Guard biocontrol strain. Genetic exchange and recombination of biocontrol strains with native strains was detected in as little as three months after biocontrol application and up to one and three years later. There was limited inter-lineage migration in the untreated fields. These findings suggest that biocontrol products that include strains from lineage IB offer the greatest potential for sustained reductions in aflatoxin levels over several years. This knowledge has important implications for developing new biocontrol strategies.

Meta-analysis of yield response of foliar fungicide-treated hybrid corn in the United States and Ontario, Canada.

BACKGROUND: Foliar fungicide applications to corn (Zea mays L.) occur at one or more application timings ranging from early vegetative growth stages to mid-reproductive stages. Previous studies indicated that fungicide applications are profitable under high disease pressure when applied during the tasseling to silking growth stages. Few comprehensive studies in corn have examined the impact of fungicide applications at an early vegetative growth stage (V6) compared to late application timings (VT) for yield response and return on fungicide investment (ROI) across multiple locations. OBJECTIVE: Compare yield response of fungicide application timing across multiple fungicide classes and calculate the probability of positive ROI. METHODS: Data were collected specifically for this analysis using a uniform protocol conducted in 13 states in the United States and one province in Canada from 2014-2015. Data were subjected to a primary mixed-model analysis of variance. Subsequent univariate meta-analyses, with and without moderator variables, were performed using standard meta-analytic procedures. Follow-up power and prediction analyses were performed to aid interpretation and development of management recommendations. RESULTS: Fungicide application resulted in a range of yield responses from -2,683.0 to 3,230.9 kg/ha relative to the non-treated control, with 68.2% of these responses being positive. Evidence suggests that all three moderator variables tested (application timing, fungicide class, and disease base level), had some effect (α = 0.05) on the absolute difference in yield between fungicide treated and non-treated plots ([Formula: see text]). Application timing influenced [Formula: see text], with V6 + VT and the VT application timings resulting in greater yield responses than the V6 application timing alone. Fungicide formulations that combined demethylation inhibitor and quinone outside inhibitor fungicides significantly increased yield response. CONCLUSION: Foliar fungicide applications can increase corn grain yield. To ensure the likelihood of a positive ROI, farmers should focus on applications at VT and use fungicides that include a mix of demethylation inhibitor and quinone outside inhibitor active ingredients.

Larval western bean cutworm feeding damage encourages the development of Gibberella ear rot on field corn.

BACKGROUND: A 2 year study was conducted to determine whether western bean cutworm (Striacosta albicosta Smith) (WBC) larval feeding damage increases severity of the fungal disease Gibberella ear rot [Fusarium graminearum (Schwein.) Petch] in field corn (Zea mays L.). The effect of a quinone-outside inhibiting fungicide, pyraclostrobin, on Gibberella ear rot severity and mycotoxin production, both with and without WBC pressure, was also evaluated. The impact of each variable was assessed individually and in combination to determine the effect of each upon ear disease severity. RESULTS: There was a positive correlation between the presence of WBC larvae in field corn and Gibberella ear rot severity under inoculated conditions in the 2 years of the experiment. An application of pyraclostrobin did not impact Gibberella ear rot development when applied at corn growth stage R1 (silks first emerging). CONCLUSION: Feeding damage from WBC larvae significantly increases the development of F. graminearum in field corn. We conclude that an effective integrated management strategy for Gibberella ear rot should target the insect pest first, in an effort to limit disease severity and subsequent mycotoxin production by F. graminearum in kernels. © 2016 Society of Chemical Industry.

Genetic Architecture of Charcoal Rot (Macrophomina phaseolina) Resistance in Soybean Revealed Using a Diverse Panel.

Charcoal rot (CR) disease caused by Macrophomina phaseolina is responsible for significant yield losses in soybean production. Among the methods available for controlling this disease, breeding for resistance is the most promising. Progress in breeding efforts has been slow due to the insufficient information available on the genetic mechanisms related to resistance. Genome-wide association studies (GWAS) enable unraveling the genetic architecture of resistance and identification of causal genes. The aims of this study were to identify new sources of resistance to CR in a collection of 459 diverse plant introductions from the USDA Soybean Germplasm Core Collection using field and greenhouse screenings, and to conduct GWAS to identify candidate genes and associated molecular markers. New sources for CR resistance were identified from both field and greenhouse screening from maturity groups I, II, and III. Five significant single nucleotide polymorphism (SNP) and putative candidate genes related to abiotic and biotic stress responses are reported from the field screening; while greenhouse screening revealed eight loci associated with eight candidate gene families, all associated with functions controlling plant defense response. No overlap of markers or genes was observed between field and greenhouse screenings suggesting a complex molecular mechanism underlying resistance to CR in soybean with varied response to different environments; but our findings provide useful information for advancing breeding for CR resistance as well as the genetic mechanism of resistance.