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.

Determining structures of individual RNA conformers using atomic force microscopy images and deep neural networks.

The vast percentage of the human genome is transcribed into RNA, many of which contain various structural elements and are important for functions. RNA molecules are conformationally heterogeneous and functionally dyanmics1, even when they are structured and well-folded2, which limit the applicability of methods such as NMR, crystallography, or cryo-EM. Moreover, because of the lack of a large structure RNA database, and no clear correlation between sequence and structure, approaches like AlphaFold3 for protein structure prediction, do not apply to RNA. Therefore determining the structures of heterogeneous RNA is an unmet challenge. Here we report a novel method of determining RNA three-dimensional topological structures using deep neural networks and atomic force microscopy (AFM) images of individual RNA molecules in solution. Owing to the high signal-to-noise ratio of AFM, our method is ideal for capturing structures of individual conformationally heterogeneous RNA. We show that our method can determine 3D topological structures of any large folded RNA conformers, from ~ 200 to ~ 420 residues, the size range that most functional RNA structures or structural elements fall into. Thus our method addresses one of the major challenges in frontier RNA structural biology and may impact our fundamental understanding of RNA structure.

Transcription factor expression repertoire basis for epigenetic and transcriptional subtypes of colorectal cancers.

Colorectal cancers (CRCs) form a heterogenous group classified into epigenetic and transcriptional subtypes. The basis for the epigenetic subtypes, exemplified by varying degrees of promoter DNA hypermethylation, and its relation to the transcriptional subtypes is not well understood. We link cancer-specific transcription factor (TF) expression alterations to methylation alterations near TF-binding sites at promoter and enhancer regions in CRCs and their premalignant precursor lesions to provide mechanistic insights into the origins and evolution of the CRC molecular subtypes. A gradient of TF expression changes forms a basis for the subtypes of abnormal DNA methylation, termed CpG-island promoter DNA methylation phenotypes (CIMPs), in CRCs and other cancers. CIMP is tightly correlated with cancer-specific hypermethylation at enhancers, which we term CpG-enhancer methylation phenotype (CEMP). Coordinated promoter and enhancer methylation appears to be driven by downregulation of TFs with common binding sites at the hypermethylated enhancers and promoters. The altered expression of TFs related to hypermethylator subtypes occurs early during CRC development, detectable in premalignant adenomas. TF-based profiling further identifies patients with worse overall survival. Importantly, altered expression of these TFs discriminates the transcriptome-based consensus molecular subtypes (CMS), thus providing a common basis for CIMP and CMS subtypes.

Influence of Integrated Management Strategies on Soybean Sudden Death Syndrome (SDS) Root Infection, Foliar Symptoms, Yield and Net Returns.

Three soybean field trials were conducted in Indiana to evaluate the integration of seed treatment, cultivar selection, and seeding rate on sudden death syndrome (SDS) root rot, pathogen load in the root, foliar symptoms, yield, and net return. Two soybean cultivars, one moderately resistant and one susceptible to SDS, were planted at three seeding rates (272,277 seeds/ha, 346,535 seeds/ha, and 420,792 seeds/ha). Fluopyram and pydiflumetofen seed treatments were applied to both cultivars, and the cultivars were then compared with a control. Low foliar SDS disease pressure was observed in our study. Seed treatment with either fluopyram or pydiflumetofen and the use of a moderately resistant cultivar decreased Fusarium virguliforme DNA concentration in the root relative to the control and the use of a susceptible cultivar. Fluopyram significantly reduced visual root rot severity by 8.8% and increased yield by 105 kg/ha relative to the control but was not different from pydiflumetofen. However, pydiflumetofen performed the same as the control with respect to root rot severity and yield. Findings from this study support the use of a seed treatment to protect roots from infection and the use of a moderately resistant cultivar planted at a seeding rate of 346,535 seeds/ha to protect yield and maximize net returns when a field has low foliar SDS pressure.

A transformer-based approach for early prediction of soybean yield using time-series images.

Crop yield prediction which provides critical information for management decision-making is of significant importance in precision agriculture. Traditional manual inspection and calculation are often laborious and time-consuming. For yield prediction using high-resolution images, existing methods, e.g., convolutional neural network, are challenging to model long range multi-level dependencies across image regions. This paper proposes a transformer-based approach for yield prediction using early-stage images and seed information. First, each original image is segmented into plant and soil categories. Two vision transformer (ViT) modules are designed to extract features from each category. Then a transformer module is established to deal with the time-series features. Finally, the image features and seed features are combined to estimate the yield. A case study has been conducted using a dataset that was collected during the 2020 soybean-growing seasons in Canadian fields. Compared with other baseline models, the proposed method can reduce the prediction error by more than 40%. The impact of seed information on predictions is studied both between models and within a single model. The results show that the influence of seed information varies among different plots but it is particularly important for the prediction of low yields.

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.

Determining independence and associations among various cardiovascular disease risk factors in 9-12 years old school-children: a cross sectional study.

BACKGROUND: Cardiovascular disease (CVD) risk assessment of children typically includes evaluating multiple CVD risk factors some of which tend to correlate each other. However, in older children and young adolescents, there are little data on the level of independence of CVD risk factors. The purpose of this study was to examine the relationships among various CVD risk factors to determine the level of independence of each risk factor in a sample of 5th-grade public school students. METHOD: A cross-sectional analysis of 1525 children (856 girls and 669 boys; age: 9-12 years) who participated in baseline CVD risk assessment for the (S)Partners for Heart Health program from 2010 - 2018. Thirteen CVD risk factor variables were used in the analysis and included blood lipids [low-density lipoprotein (LDL), high-density lipoprotein (HDL), total cholesterol (TC), and triglycerides], resting systolic and diastolic blood pressure (BP); anthropometrics [height, weight, body mass index (BMI), % body fat, waist circumference (WC)]. Additionally, acanthosis nigricans (a marker insulin resistance and diabetes), and cardiorespiratory fitness (VO2 ml/kg) was estimated using the PACER. Descriptive statistics, bivariate Pearson correlations, and principal component analysis were used to determine the relationships among these variables and the independence. RESULTS: Parallel analysis indicated two components should be extracted. Among the two components extracted, WC, % body fat, and BMI loaded highest on component 1, which explained 34% of the total variance. Systolic BP and diastolic BP loaded predominantly on component 2 and accounted for 17% of the variance. Cardiorespiratory fitness, acanthosis nigricans, HDL, and triglycerides loaded highest on the first component (loadings between 0.42 and 0.57) but still suggest some non-shared variance with this component. Low-density lipoprotein had low loadings on each component. Factor loadings were stable across sex. CONCLUSION: Among the various CVD risk indicators, measures of adiposity loaded highest on the component that explained the largest proportion of variability in the data reinforcing the importance of assessing adiposity in CVD risk assessment. In addition, blood pressure loaded highest on the second component, suggesting their relative independence when assessing CVD risk. The data also provide support and rationale for determining what CVD risk factors to include- based on resource needs. For example, researchers or public health programs may choose to assess WC instead of lipid profile for cardiovascular related problems if ease of assessment and cost are considerations.

Synchronous RNA conformational changes trigger ordered phase transitions in crystals.

Time-resolved studies of biomacromolecular crystals have been limited to systems involving only minute conformational changes within the same lattice. Ligand-induced changes greater than several angstroms, however, are likely to result in solid-solid phase transitions, which require a detailed understanding of the mechanistic interplay between conformational and lattice transitions. Here we report the synchronous behavior of the adenine riboswitch aptamer RNA in crystal during ligand-triggered isothermal phase transitions. Direct visualization using polarized video microscopy and atomic force microscopy shows that the RNA molecules undergo cooperative rearrangements that maintain lattice order, whose cell parameters change distinctly as a function of time. The bulk lattice order throughout the transition is further supported by time-resolved diffraction data from crystals using an X-ray free electron laser. The synchronous molecular rearrangements in crystal provide the physical basis for studying large conformational changes using time-resolved crystallography and micro/nanocrystals.

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.

Neonicotinoid seed treatments of soybean provide negligible benefits to US farmers.

Neonicotinoids are the most widely used insecticides worldwide and are typically deployed as seed treatments (hereafter NST) in many grain and oilseed crops, including soybeans. However, there is a surprising dearth of information regarding NST effectiveness in increasing soybean seed yield, and most published data suggest weak, or inconsistent yield benefit. The US is the key soybean-producing nation worldwide and this work includes soybean yield data from 194 randomized and replicated field studies conducted specifically to evaluate the effect of NSTs on soybean seed yield at sites within 14 states from 2006 through 2017. Here we show that across the principal soybean-growing region of the country, there are negligible and management-specific yield benefits attributed to NSTs. Across the entire region, the maximum observed yield benefits due to fungicide (FST = fungicide seed treatment) + neonicotinoid use (FST + NST) reached 0.13 Mg/ha. Across the entire region, combinations of management practices affected the effectiveness of FST + NST to increase yield but benefits were minimal ranging between 0.01 to 0.22 Mg/ha. Despite widespread use, this practice appears to have little benefit for most of soybean producers; across the entire region, a partial economic analysis further showed inconsistent evidence of a break-even cost of FST or FST + NST. These results demonstrate that the current widespread prophylactic use of NST in the key soybean-producing areas of the US should be re-evaluated by producers and regulators alike.

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.

Aging-like Spontaneous Epigenetic Silencing Facilitates Wnt Activation, Stemness, and BrafV600E-Induced Tumorigenesis.

We addressed the precursor role of aging-like spontaneous promoter DNA hypermethylation in initiating tumorigenesis. Using mouse colon-derived organoids, we show that promoter hypermethylation spontaneously arises in cells mimicking the human aging-like phenotype. The silenced genes activate the Wnt pathway, causing a stem-like state and differentiation defects. These changes render aged organoids profoundly more sensitive than young ones to transformation by BrafV600E, producing the typical human proximal BRAFV600E-driven colon adenocarcinomas characterized by extensive, abnormal gene-promoter CpG-island methylation, or the methylator phenotype (CIMP). Conversely, CRISPR-mediated simultaneous inactivation of a panel of the silenced genes markedly sensitizes to BrafV600E-induced transformation. Our studies tightly link aging-like epigenetic abnormalities to intestinal cell fate changes and predisposition to oncogene-driven colon tumorigenesis.

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.

Differences in Small Plot and On-Farm Trials for Yield Response to Foliar Fungicide in Soybean.

Soybean yield response variability to foliar fungicide applications was evaluated in on-farm replicated strip trials (OFTs) and small-plot trials (SPTs) from 2008 through 2015 in Iowa. A total of 230 OFTs and 49 SPTs were compared for yield response to pyraclostrobin, pyraclostrobin + fluxapyroxad, or trifloxystrobin + prothioconazole fungicides. OFTs (18 to 55 m wide and 200 to 800 m long strips) were harvested with farmers' combines equipped with yield monitors and GPS, while SPTs (3.0 to 4.6 m wide and 10.7 to 15.3 m long plots) were harvested by small research plot combines. Variance component and power analyses were conducted with a subset of data consisting of 12 OFTs and SPTs, each with pyraclostrobin and evaluated in 2008 and 2009. While average yield responses were similar, the residual random yield variation was smaller in OFTs than SPTs. Power analysis showed that SPTs need more replications than OFTs to detect the same overall treatment differences. To detect a yield response of 134 kg/ha, it would require at least three treatment replications with 12 locations in OFTs and seven replications with 12 locations in SPTs. Researchers need to acknowledge the differences in statistical power of detecting yield responses to foliar fungicide on soybean in different types of field experiments, especially with smaller plot sizes in situations with less foliar disease.

Topological Structure Determination of RNA Using Small-Angle X-Ray Scattering.

Knowledge of RNA three-dimensional topological structures provides important insight into the relationship between RNA structural components and function. It is often likely that near-complete sets of biochemical and biophysical data containing structural restraints are not available, but one still wants to obtain knowledge about approximate topological folding of RNA. In this regard, general methods for determining such topological structures with minimum readily available restraints are lacking. Naked RNAs are difficult to crystallize and NMR spectroscopy is generally limited to small RNA fragments. By nature, sequence determines structure and all interactions that drive folding are self-contained within sequence. Nevertheless, there is little apparent correlation between primary sequences and three-dimensional folding unless supplemented with experimental or phylogenetic data. Thus, there is an acute need for a robust high-throughput method that can rapidly determine topological structures of RNAs guided by some experimental data. We present here a novel method (RS3D) that can assimilate the RNA secondary structure information, small-angle X-ray scattering data, and any readily available tertiary contact information to determine the topological fold of RNA. Conformations are firstly sampled at glob level where each glob represents a nucleotide. Best-ranked glob models can be further refined against solvent accessibility data, if available, and then converted to explicit all-atom coordinates for refinement against SAXS data using the Xplor-NIH program. RS3D is widely applicable to a variety of RNA folding architectures currently present in the structure database. Furthermore, we demonstrate applicability and feasibility of the program to derive low-resolution topological structures of relatively large multi-domain RNAs.

Real-time crystallographic studies of the adenine riboswitch using an X-ray free-electron laser.

Structures of the four reaction states of the adenine riboswitch aptamer domain, including a transient intermediate state were solved by serial femtosecond crystallography. The structures not only demonstrate the use of X-ray free-electron lasers for RNA crystallography but have also proven that transient states can be determined in real time by mix-and-inject crystallography. These results illustrate the structural basis for the ligand-induced conformational changes associated with the molecular 'switch'.