Management of Soybean Cyst Nematode and Sudden Death Syndrome with Nematode-Protectant Seed Treatments Across Multiple Environments in Soybean.

As soybean (Glycine max) production continues to expand in the United States and Canada, so do pathogens and pests that directly threaten soybean yield potential and economic returns for farmers. One such pathogen is the soybean cyst nematode (SCN; Heterodera glycines). SCN has traditionally been managed using SCN-resistant cultivars and rotation with nonhost crops, but the interaction of SCN with sudden death syndrome (SDS; caused by Fusarium virguliforme) in the field makes management more difficult. Nematode-protectant seed treatments have become options for SCN and SDS management. The objectives of this study were to evaluate nematode-protectant seed treatments for their effects on (i) early and full season SCN reproduction, (ii) foliar symptoms and root-rot caused by SDS, and (iii) soybean yield across environments accounting for the above factors. Using a standard protocol, field trials were implemented in 13 states and one Canadian province from 2019 to 2021 constituting 51 site-years. Six nematode-protectant seed treatment products were compared with a fungicide + insecticide base treatment and a nontreated check. Initial (at soybean planting) and final (at soybean harvest) SCN egg populations were enumerated, and SCN females were extracted from roots and counted at 30 to 35 days postplanting. Foliar disease index (FDX) and root rot caused by the SDS pathogen were evaluated, and yield data were collected for each plot. No seed treatment offered significant nematode control versus the nontreated check for in-season and full-season nematode response, no matter the initial SCN population or FDX level. Of all treatments, ILEVO (fluopyram) and Saltro (pydiflumetofen) provided more consistent increases in yield over the nontreated check in a broader range of SCN environments, even when FDX level was high.

A new and effective method to induce infection of Phyllachora maydis into corn for tar spot studies in controlled environments.

BACKGROUND: Tar spot of corn is a significant and spreading disease in the continental U.S. and Canada caused by the obligate biotrophic fungus Phyllachora maydis. As of 2023, tar spot had been reported in 18 U.S. states and one Canadian Province. The symptoms of tar spot include chlorotic flecking followed by the formation of black stromata where conidia and ascospores are produced. Advancements in research and management for tar spot have been limited by a need for a reliable method to inoculate plants to enable the study of the disease. The goal of this study was to develop a reliable method to induce tar spot in controlled conditions. RESULTS: We induced infection of corn by P. maydis in 100% of inoculated plants with a new inoculation method. This method includes the use of vacuum-collection tools to extract ascospores from field-infected corn leaves, application of spores to leaves, and induction of the disease in the dark at high humidity and moderate temperatures. Infection and disease development were consistently achieved in four independent experiments on different corn hybrids and under different environmental conditions in a greenhouse and growth chamber. Disease induction was impacted by the source and storage conditions of spores, as tar spot was not induced with ascospores from leaves stored dry at 25 ºC for 5 months but was induced using ascospores from infected leaves stored at -20 ºC for 5 months. The time from inoculation to stromata formation was 10 to 12 days and ascospores were present 19 days after inoculation throughout our experiments. In addition to providing techniques that enable in-vitro experimentation, our research also provides fundamental insights into the conditions that favor tar spot epidemics. CONCLUSIONS: We developed a method to reliably inoculate corn with P. maydis. The method was validated by multiple independent experiments in which infection was induced in 100% of the plants, demonstrating its consistency in controlled conditions. This new method facilitates research on tar spot and provides opportunities to study the biology of P. maydis, the epidemiology of tar spot, and for identifying host resistance.

Is Microdochium maydis Associated with Necrotic Lesions in the Tar Spot Disease Complex? A Culture-Based Survey of Maize in Mexico and the Midwestern United States.

Tar spot, caused by Phyllachora maydis, is an emerging disease of corn in the United States. Stromata of P. maydis are sometimes surrounded by necrotic lesions known as fisheyes and were previously reported to be caused by the fungus Microdochium maydis. The association of M. maydis with fisheye lesions has not been well documented outside of initial descriptions from the early 1980s. The objective of this work was to assess and identify Microdochium-like fungi associated with necrotic lesions surrounding P. maydis stromata using a culture-based method. In 2018, corn leaf samples with fisheye lesions associated with tar spot stromata were collected from 31 production fields across Mexico, Illinois, and Wisconsin. Cultures of pure isolates collected from Mexico believed to be M. maydis were included in the study. A total of 101 Microdochium/Fusarium-like isolates were obtained from the necrotic lesions, and 91% were identified as Fusarium spp., based on initial ITS sequence data. Multi-gene (ITS, TEF1-α, RPB1, and RPB2) phylogenies were constructed for a subset of 55 isolates; Microdochium, Cryptostroma, and Fusarium reference sequences were obtained from GenBank. All the necrotic lesion isolates clustered within Fusarium lineages and were phylogenetically distinct from the Microdochium clade. All Fusarium isolates from Mexico belonged to the F. incarnatum-equiseti species complex, whereas >85% of the U.S. isolates grouped within the F. sambucinum species complex. Our study suggests that initial reports of M. maydis were misidentifications of resident Fusarium spp. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Efficacy and Profitability of Fungicides for Managing Frogeye Leaf Spot on Soybean in the United States: A 10-Year Quantitative Summary.

Frogeye leaf spot (FLS), caused by Cercospora sojina, is an economically important disease of soybean in the United States. Data from 66 uniform fungicide trials (UFTs) conducted from 2012 to 2021 across eight states (Alabama, Arkansas, Illinois, Iowa, Kentucky, Louisiana, Mississippi, and Tennessee) were gathered and analyzed to determine the efficacy and profitability of the following fungicides applied at the beginning pod developmental stage (R3): azoxystrobin + difenoconazole (AZOX + DIFE), difenoconazole + pydiflumetofen (DIFE + PYDI), pyraclostrobin (PYRA), pyraclostrobin + fluxapyroxad + propiconazole (PYRA + FLUX + PROP), tetraconazole (TTRA), thiophanate-methyl (TMET), thiophanate-methyl + tebuconazole (TMET + TEBU), and trifloxystrobin + prothioconazole (TFLX + PROT). A network meta-analytic model was fitted to the log of the means of FLS severity data and to the nontransformed mean yield for each treatment, including the nontreated. The percent reduction in disease severity (%) and the yield response (kg/ha) relative to the nontreated was the lowest for PYRA (11%; 136 kg/ha) and the greatest for DIFE + PYDI (57%; 441 kg/ha). A significant decline in efficacy over time was detected for PYRA (18 percentage points [p.p.]), TTRA (27 p.p.), AZOX + DIFE (18 p.p.), and TMET + TEBU (19 p.p.) by using year as a continuous covariate in the model. Finally, probabilities of breaking even were the greatest (>65%) for the most effective fungicide DIFE + PYDI and the lowest (<55%) for PYRA. Results of this meta-analysis may be useful to support decisions when planning fungicide programs.

Meta-Analytic Modeling of the Severity-Yield Relationships in Soybean Frogeye Leaf Spot Epidemics.

Frogeye leaf spot (FLS), caused by Cercospora sojina, is an important foliar disease affecting soybean in the United States. A meta-analytic approach including 39 fungicide trials conducted from 2012 to 2021 across eight states (Alabama, Arkansas, Illinois, Iowa, Kentucky, Louisiana, Mississippi, Tennessee) was used to assess the relationship between FLS severity and soybean yield. Correlation and regression analyses were performed separately to determine Fisher's transformation of correlation coefficients (Zr), intercept (ß0) and slope (ß1). Disease pressure (low severity, ≤34.5; high severity, >34.5%) and yield class (low, ≤3,352; high, >3,352 kg/ha) were included as categorical moderators. Pearson's [Formula: see text], obtained from back-transforming the [Formula: see text]r estimated by an overall random-effects model, showed a significant negative linear relationship between FLS severity and yield ([Formula: see text] = -0.60). The [Formula: see text]r was affected by disease pressure (P = 0.0003) but not by yield class (P = 0.8141). A random-coefficient model estimated a slope of -19 kg/ha for each percent severity for a mean attainable yield of 3,719.9 kg/ha. Based on the overall mean (95% CI) of the intercept and slope estimated by the random-coefficients model, the estimated overall relative damage coefficient was 0.51% (0.36 to 0.69), indicating that a percent increase in FLS severity reduced yield by 0.51%. The best model included yield class as a covariate, and population-average intercepts differed significantly between low (3,455.1 kg/ha) and high (3,842.7 kg/ha) yield classes. This highlights the potential impact of FLS on soybean yield if not managed and may help in disease management decisions.

Field and Greenhouse Assessment of Seed Treatment Fungicides for Management of Sudden Death Syndrome and Yield Response of Soybean.

Seed treatments for the management of sudden death syndrome (SDS) caused by Fusarium virguliforme are available in the United States and Canada; however, side-by-side comparisons of these seed treatments are lacking. Sixteen field experiments were established in Illinois, Indiana, Iowa, Michigan, and Wisconsin, United States, and Ontario, Canada, in 2019 and 2020 to evaluate seed treatment combinations. Treatments included a nontreated check (NTC), fungicide and insecticide base seed treatments (base), fluopyram, base + fluopyram, base + saponin extracts from Chenopodium quinoa, base + fluopyram + heat-killed Burkholderia rinojenses, base + pydiflumetofen, base + thiabendazole + heat-killed B. rinojenses, and base + thiabendazole + C. quinoa extracts + heat-killed B. rinojenses. Treatments were tested on SDS moderately resistant and susceptible soybean cultivars at each location. Overall, NTC and base had the most root rot, most foliar disease index (FDX), and lowest yield. Base + fluopyram and base + pydiflumetofen were most effective for managing SDS. Moderately resistant cultivars reduced FDX in both years but visual root rot was greater on the moderately resistant than the susceptible cultivars in 2020. Yield response to cultivar was also inconsistent between the 2 years. In 2020, the susceptible cultivar provided significantly more yield than the moderately resistant cultivar. Treatment effect for root rot and FDX was similar in field and greenhouse evaluations. These results reinforce the need to include root rot evaluations in addition to foliar disease evaluations in the breeding process for resistance to F. virguliforme and highlights the importance of an integrated SDS management plan because not a single management tactic alone provides adequate control of the disease.

Virulence, Aggressiveness, and Fungicide Sensitivity of Phytophthora spp. Associated with Soybean in Illinois.

Phytophthora root and stem rot (PRR), caused by Phytophthora sojae, is one of the most devastating oomycete diseases of soybean in Illinois. Single resistant genes (Rps) are used to manage this pathogen but P. sojae has adapted to Rps, causing failure of resistance in many regions. In addition to P. sojae, recent reports indicate that P. sansomeana could also cause root rot in soybean. Soil samples and symptomatic plants were collected across 40 Illinois counties between 2016 and 2018. P. sojae (77%) was more abundant than P. sansomeana (23%) across Illinois fields. Both species were characterized by virulence, aggressiveness, and fungicide sensitivity. Virulence of all P. sojae isolates was evaluated using the hypocotyl inoculation technique in 13 soybean differentials. Aggressiveness was evaluated in the greenhouse by inoculating a susceptible cultivar and measuring root and shoot dry weight. On average, P. sojae isolates were able to cause disease on six soybean differentials. P. sojae was more aggressive than P. sansomeana. All isolates were sensitive to azoxystrobin, ethaboxam, mefenoxam, and metalaxyl. The characterization of the population of species associated with PRR will inform management decisions for this disease in Illinois.

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.

Phyllachora species infecting maize and other grass species in the Americas represents a complex of closely related species.

The genus Phyllachora contains numerous obligate fungal parasites that produce raised, melanized structures called stromata on their plant hosts referred to as tar spot. Members of this genus are known to infect many grass species but generally do not cause significant damage or defoliation, with the exception of P. maydis which has emerged as an important pathogen of maize throughout the Americas, but the origin of this pathogen remains unknown. To date, species designations for Phyllachora have been based on host associations and morphology, and most species are assumed to be host specific. We assessed the sequence diversity of 186 single stroma isolates collected from 16 hosts representing 15 countries. Samples included both herbarium and contemporary strains that covered a temporal range from 1905 to 2019. These 186 isolates were grouped into five distinct species with strong bootstrap support. We found three closely related, but genetically distinct groups of Phyllachora are capable of infecting maize in the United States, we refer to these as the P. maydis species complex. Based on herbarium specimens, we hypothesize that these three groups in the P. maydis species complex originated from Central America, Mexico, and the Caribbean. Although two of these groups were only found on maize, the third and largest group contained contemporary strains found on maize and other grass hosts, as well as herbarium specimens from maize and other grasses that include 10 species of Phyllachora. The herbarium specimens were previously identified based on morphology and host association. This work represents the first attempt at molecular characterization of Phyllachora species infecting grass hosts and indicates some Phyllachora species can infect a broad range of host species and there may be significant synonymy in the Phyllachora genus.

Contour-Based Detection and Quantification of Tar Spot Stromata Using Red-Green-Blue (RGB) Imagery.

Quantifying symptoms of tar spot of corn has been conducted through visual-based estimations of the proportion of leaf area covered by the pathogenic structures generated by Phyllachora maydis (stromata). However, this traditional approach is costly in terms of time and labor, as well as prone to human subjectivity. An objective and accurate method, which is also time and labor-efficient, is of an urgent need for tar spot surveillance and high-throughput disease phenotyping. Here, we present the use of contour-based detection of fungal stromata to quantify disease intensity using Red-Green-Blue (RGB) images of tar spot-infected corn leaves. Image blocks (n = 1,130) generated by uniform partitioning the RGB images of leaves, were analyzed for their number of stromata by two independent, experienced human raters using ImageJ (visual estimates) and the experimental stromata contour detection algorithm (SCDA; digital measurements). Stromata count for each image block was then categorized into five classes and tested for the agreement of human raters and SCDA using Cohen's weighted kappa coefficient (κ). Adequate agreements of stromata counts were observed for each of the human raters to SCDA (κ = 0.83) and between the two human raters (κ = 0.95). Moreover, the SCDA was able to recognize "true stromata," but to a lesser extent than human raters (average median recall = 90.5%, precision = 89.7%, and Dice = 88.3%). Furthermore, we tracked tar spot development throughout six time points using SCDA and we obtained high agreement between area under the disease progress curve (AUDPC) shared by visual disease severity and SCDA. Our results indicate the potential utility of SCDA in quantifying stromata using RGB images, complementing the traditional human, visual-based disease severity estimations, and serve as a foundation in building an accurate, high-throughput pipeline for the scoring of tar spot symptoms.

Diaporthe Seed Decay of Soybean [Glycine max (L.) Merr.] Is Endemic in the United States, But New Fungi Are Involved.

Diaporthe seed decay can compromise seed quality in soybean [Glycine max (L.) Merr.] in the warm and humid production areas of the United States during crop maturation. In the current study, 45 isolates of Diaporthe were recovered from seed sampled from soybean fields affected by Diaporthe-associated diseases in eight U.S. states in 2017. The isolates obtained belonged to 10 species of Diaporthe based on morphology and phylogenetic analyses of the internal transcribed spacer, partial translation elongation factor 1-α, and ß-tubulin gene sequences. The associated species included D. aspalathi, D. caulivora, D. kongii, D. longicolla, D. sojae, D. ueckerae, D. unshiuensis, and three novel fungi, D. bacilloides, D. flavescens, and D. insulistroma. One isolate each of the 10 species was examined for pathogenicity on seed of cultivar Sava under controlled conditions. Seven days postinoculation, significant differences in the percentages of decayed seeds and seedling necrosis were observed among the isolates and the noninoculated control (P < 0.0001). While the isolates of D. bacilloides, D. longicolla, and D. ueckerae caused a significantly greater percentage of decayed seeds (P < 0.0001), the isolate of D. aspalathi caused the greatest seedling necrosis (P < 0.0001). The observation of new fungi causing Diaporthe seed decay suggests the need for a more comprehensive survey in U.S. soybean producing areas since members of the genus Diaporthe appear to form a complex that causes seed decay.

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.

Occurrence of Quinone Outside Inhibitor Resistance in Virginia Populations of Parastagonospora nodorum Infecting Wheat.

Stagonospora nodorum blotch (SNB) of wheat, caused by Parastagonospora nodorum, is managed using cultural practices, resistant varieties, and foliar fungicides. Frequent fungicide use can select for fungicide resistance, making certain chemistries less effective; this may in part explain the increasing severity of SNB in the mid-Atlantic United States. Quinone outside inhibitor (QoI) resistance has been documented for a diversity of fungi, but it has not been reported for P. nodorum in the United States. The objectives of this study were (i) to evaluate QoI sensitivity of P. nodorum from Virginia wheat fields, (ii) to screen P. nodorum for QoI target site mutations in the cytochrome b gene, and (iii) to develop a molecular assay to detect target site mutations associated with QoI resistance. Sensitivity of 16 isolates to pyraclostrobin and azoxystrobin was evaluated with radial growth assays, and the cytochrome b gene was sequenced. One isolate was insensitive to both fungicides and had the G143A mutation in the cytochrome b gene. For azoxystrobin, 10 isolates without target site mutations had reduced sensitivity. Additional isolates (n = 58) were sequenced. A total of seven isolates had the G143A mutation and also had reduced sensitivity to pyraclostrobin and azoxystrobin compared with a sensitive control isolate without the mutation. A pyrosequencing assay targeting G143A was developed as a rapid method to screen P. nodorum for the QoI resistance-conferring mutation. To our knowledge, this is the first report of QoI-resistant P. nodorum in the United States. Overall resistance frequency was low, but resistance management practices are needed to maintain the efficacy of fungicides for SNB control.

Assessing the Curative and Protective Impacts of Select Fungicides for Control of Powdery Mildew of Wheat.

Powdery mildew, caused by the obligate fungal pathogen Blumeria graminis, has been increasing in incidence and severity on wheat in the Chesapeake Bay region of the mid-Atlantic United States. Although fungicides are used for in-season management of powdery mildew, no studies to date have assessed the ability of these products to control disease after pathogen arrival/infection (curative activity) and the duration of disease control provided after fungicide application (protective activity) under controlled conditions. Five commercially available fungicide products, including Caramba, Stratego YLD, Priaxor, Prosaro, and Trivapro, were applied at either 3 or 5 days after inoculation with B. graminis spores to assess curative activity. In a separate study, protective activity for these fungicides was assessed by applying fungicides and inoculating with B. graminis spores the same day and 21 and 42 days after fungicide application. All fungicides reduced powdery mildew severity on foliage compared with nontreated controls. Priaxor provided 11 to 18% less activity than other fungicides when applied curatively. All products provided protective control up to 42 days after inoculation, with Stratego YLD and Priaxor providing the greatest level of control at 68 and 56%, respectively. Our data indicate that different fungicides can have subtle differences in overall efficacy profiles that may translate to improved control or an extended control window in some situations, but all tested products provide very good to excellent control of powdery mildew on wheat.

Relationship Between Invasive Brown Marmorated Stink Bug (Halyomorpha halys) and Fumonisin Contamination of Field Corn in the Mid-Atlantic U.S.

Brown marmorated stink bug (Halyomorpha halys Stål) is an invasive agricultural pest that causes severe damage to many crops. To determine potential associations between H. halys feeding damage, Fusarium infection, and mycotoxin contamination in field corn, a field survey was conducted in eight counties in Virginia. Results indicated an association between H. halys feeding damage and fumonisin contamination. Subsequent field experiments in Delaware, Maryland, and Virginia examined the ability of H. halys to increase Fusarium verticillioides (Sacc.) Nirenberg infection and fumonisin concentrations in corn. At the milk stage, H. halys (0 or 4 adults) and Fusarium (with or without F. verticillioides inoculum) treatments were applied to bagged ears in a two by two factorial randomized complete block design with 12 replicates. H. halys treatments increased levels of feeding damage (P < 0.0001) and Fusarium infection (P = 0.0380). Interaction between H. halys and Fusarium treatments influenced severity of infection (P = 0.0018) and fumonisin concentrations (P = 0.0360). Results suggest H. halys has the ability to increase both Fusarium infection and fumonisin concentrations in field corn. Further studies are needed to understand mechanisms by which H. halys increases fumonisin and to develop management strategies to mitigate impacts of H. halys on field corn in the region.

Meta-Analytic and Economic Approaches for Evaluation of Pesticide Impact on Sclerotinia Stem Rot Control and Soybean Yield in the North Central United States.

As complete host resistance in soybean has not been achieved, Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum continues to be of major economic concern for farmers. Thus, chemical control remains a prevalent disease management strategy. Pesticide evaluations were conducted in Illinois, Iowa, Michigan, Minnesota, New Jersey, and Wisconsin from 2009 to 2016, for a total of 25 site-years (n = 2,057 plot-level data points). These studies were used in network meta-analyses to evaluate the impact of 10 popular pesticide active ingredients, and seven common application timings on SSR control and yield benefit, compared with not treating with a pesticide. Boscalid and picoxystrobin frequently offered the best reductions in disease severity and best yield benefit (P < 0.0001). Pesticide applications (one- or two-spray programs) made during the bloom period provided significant reductions in disease severity index (DIX) (P < 0.0001) and led to significant yield benefits (P = 0.0009). Data from these studies were also used in nonlinear regression analyses to determine the effect of DIX on soybean yield. A three-parameter logistic model was found to best describe soybean yield loss (pseudo-R2 = 0.309). In modern soybean cultivars, yield loss due to SSR does not occur until 20 to 25% DIX, and considerable yield loss (-697 kg ha-1 or -10 bu acre-1) is observed at 68% DIX. Further analyses identified several pesticides and programs that resulted in greater than 60% probability for return on investment under high disease levels.

Evaluating the Profitability of Foliar Fungicide Programs in Mid-Atlantic Soft-Red Winter Wheat Production.

In mid-Atlantic soft-red winter wheat (SRWW) production, the standard timing for a fungicide application is between flag leaf emergence (Feekes growth stage [FGS] 8) and heading (FGS 10.5). However, two-pass and anthesis (FGS 10.5.1) applications are becoming common, although these programs have not been thoroughly evaluated for disease control, yield, and profitability. Experiments were conducted in the mid-Atlantic in 2015 and 2016 to evaluate fungicide programs with applications at FGS 8, FGS 10.5.1, and two-pass programs with an early application at green-up (FGS 5) followed by (FB) applications at either FGS 8 or FGS 10.5.1. Fungicide programs that included an application at FGS 10.5.1 resulted in the highest probability of no disease on the flag leaf (0.29 to 0.40). The estimated mean yield increases ( D¯ ) relative to the nontreated check ranged from 253.65 to 634.16 kg ha-1. Using a grain price of $0.18 kg-1 ($5 bushel-1), probabilities were similar between applications at FGS 8 (0.49 to 0.56) and FGS 10.5.1 (0.53). The probability of profitability ranged from 0.48 to 0.57 for FGS 5 FB FGS 8 applications and 0.52 to 0.59 for FGS 5 FB FGS 10.5.1 applications, indicating limited benefit to two-pass programs.

Nitrogen-fixing bacteria, arbuscular mycorrhizal fungi, and the productivity and structure of prairie grassland communities.

Due to their complementary roles in meeting plant nutritional needs, arbuscular mycorrhizal fungi (AMF) and nitrogen-fixing bacteria (N(2)-fixers) may have synergistic effects on plant communities. Using greenhouse microcosms, we tested the effects of AMF, N(2)-fixers (symbiotic: rhizobia, and associative: Azospirillum brasilense), and their potential interactions on the productivity, diversity, and species composition of diverse tallgrass prairie communities and on the productivity of Panicum virgatum in monoculture. Our results demonstrate the importance of AMF and N(2)-fixers as drivers of plant community structure and function. In the communities, we found a positive effect of AMF on diversity and productivity, but a negative effect of N(2)-fixers on productivity. Both AMF and N(2)-fixers affected relative abundances of species. AMF shifted the communities from dominance by Elymus canadensis to Sorghastrum nutans, and seven other species increased in abundance with AMF, accounting for the increased diversity. N(2)-fixers led to increases in Astragalus canadensis and Desmanthus illinoense, two legumes that likely benefited from the presence of the appropriate rhizobia symbionts. Sorghastrum nutans declined 44 % in the presence of N(2)-fixers, with the most likely explanation being increased competition from legumes. Panicum monocultures were more productive with AMF, but showed no response to N(2)-fixers, although inference was constrained by low Azospirillum treatment effectivity. We did not find interactions between AMF and N(2)-fixers in communities or Panicum monocultures, indicating that short-term effects of these microbial functional groups are additive.

Effects of soil type, fertilization and drought on carbon allocation to root growth and partitioning between secondary metabolism and ectomycorrhizae of Betula papyrifera.

Paper birch (Betula papyrifera Marsh) seedlings were grown in a greenhouse in either subsoil or topsoil in factorial combination with two fertilization and drought regimes to investigate how different soil environments and nutrient availability drive belowground partitioning between growth, secondary metabolism and ectomycorrhizal (EM) associations, and impact drought tolerance of paper birch. Root and total seedling dry biomass, starch, soluble sugars, soluble phenolics, lignin and EM abundance were quantified. In unfertilized topsoil, total plant biomass and root biomass were approximately nine times higher than in unfertilized subsoil, but the root weight ratios did not differ between soils. Root soluble phenolics and lignin were higher in unfertilized subsoil than in unfertilized topsoil, whereas EM abundance was significantly higher in unfertilized topsoil than in unfertilized subsoil. In topsoil, fertilization decreased root biomass and EM abundance and increased root phenolics and lignin. In contrast, fertilization of subsoil increased root biomass but decreased root phenolics and lignin, while EM abundance was unaffected. In both soil types, fertilization reduced root weight ratios. Across soil types, EM abundance was negatively correlated with root soluble sugars, root phenolics and lignin, but this was driven mainly by the responses in the topsoil treatment. Our results show that soil fertility mediates carbon tradeoffs among defense, growth and EM associations.

Leaf Blight Disease on the Invasive Grass Microstegium vimineum Caused by a Bipolaris sp.

In 2009, a previously undescribed disease was found on the nonnative invasive annual grass Microstegium vimineum (Japanese stiltgrass). Diseased plants exhibited foliar lesions, wilting, and in some cases, death of entire plants. We identified the causal agent as a Bipolaris sp. similar to B. zeicola. We observed spores and associated structures characteristic of Bipolaris spp. growing from leaf lesions on field collected plants. Pure cultures of the fungus were made and spore suspensions were applied to laboratory-reared M. vimineum seedlings in growth chamber and greenhouse experiments. Initial symptoms appeared on seedlings in the growth chamber experiment within 72 h of inoculation, and seedlings exhibited characteristic lesions within 10 days. The fungus was reisolated from lesions, and the internal transcribed spacer (ITS) region was sequenced to confirm its identity. In the greenhouse experiment, inoculated plants displayed characteristic lesions, and relatively greater spore loads increased disease incidence. Disease reduced seed head production by 40% compared to controls. This is the first report of a Bipolaris sp. causing disease on invasive M. vimineum. Following further analysis, including assays with co-occurring native species, this Bipolaris sp. may be considered as a biocontrol agent for invasive M. vimineum.