Biology subjects

Gage, J. L.

Publications and source records attributed to Gage, J. L..

12 recordsLinked to original sources

bioRxiv2026

HyARQ: a Snakemake workflow for allele-specific read counting and quality control in F1 hybrids

Summary Allele-specific analysis in F1 hybrids allows direct assessment of how cis-regulatory variation contributes to differences in gene expression and other regulatory features. However, allele-specific analyses are sensitive to errors arising from multiple sources, including mapping bias and technical variation, requiring careful processing and quality control. These challenges are compounded by the fact that workflows for different molecular assays are often developed separately, making consistent analysis and quality control across assays difficult. Here, we introduce HyARQ, a Snakemake workflow for allele-specific read counting and quality control across RNA-seq, MOA-seq, ATAC-seq, and ChIP-seq data. HyARQ integrates assay-specific preprocessing, mapping to concatenated parental genomes, filtering, counting at paired parental genes or regulatory regions, and quality control within a common workflow. Using simulated datasets with known parental origin and allelic proportions, we show that HyARQ accurately recovers expected parental proportions with minimal cross-parent assignment across all four assay types. We further applied HyARQ to real RNA-seq, MOA-seq, ATAC-seq, and ChIP-seq datasets and generated paired parental counts and assay-specific quality control outputs for each assay. HyARQ provides a standardized and reproducible framework for allele-specific analysis across multiple molecular layers and facilitates downstream studies of cis-regulatory variation in F1 hybrids.

bioinformatics↗

bioRxiv2026

Cis-regulatory variation and transcription factor binding contribute to allelic genotype-by-environment interactions for gene expression in maize

Genotype-by-environment interactions (GxE), or differences in how genotypes perform across varying environments, are a pervasive source of phenotypic variation and underlie differences in local adaptation. Though GxE is well characterized across kingdoms of life, less is known about what causes GxE interactions, particularly at the molecular level. In this study, we use allele-specific gene expression estimates in a maize (Zea mays L.) B73 x Mo17 hybrid to isolate cis-regulatory effects on gene expression for each of the two parental alleles. The hybrid was grown in two environments, and expression differences between the parental alleles were used to characterize allele-by-environment (AxE) interactions and study the influence of gene-proximal sequence variation on transcript abundance AxE. We tested the hypothesis that gene-proximal sequence variation can cause GxE in gene expression by modifying transcription factor binding. Our results show that sequence variation in gene promoter regions has a small but consistent enrichment in genes that show transcriptional AxE. Further, we demonstrate that differential transcription factor binding potential caused by sequence variation is also enriched in AxE genes. Predictive models trained on sequence and transcription factor binding variation show that while these features contain some information about whether a gene will show transcriptional AxE, they alone are not sufficient to reliably distinguish AxE genes. These findings support the hypothesis that gene expression GxE can be caused by sequence variation that modifies transcription factor binding, while also reinforcing the complex and context-specific nature of GxE interactions.

genomics↗

bioRxiv2026

Aerial imagery and deep learning accurately estimate maize foliar disease severity

Southern leaf blight (SLB) is a foliar disease of maize (Zea mays L.) caused by the necrotrophic fungal pathogen Cochliobolus heterostrophus. Genetic resistance is the most effective control method for SLB. Developing disease resistant maize lines requires field trials during which disease phenotypes must be visually assessed. Remote sensing using drones is an emerging technology that can be leveraged for high-throughput phenotyping of disease severity that is otherwise labor-intensive and subjective. This project used a deep learning approach to estimate SLB disease severity of single-row maize plots from drone imagery. Over 26,000 plot-level images produced from flights conducted across three growing seasons were labeled with in-field visual scores taken contemporaneously by expert raters. Variation in environmental conditions contributed to a labeled image dataset that reflects the complexity of agronomic field experiments. We assessed the ability of nine deep learning models from three architectural families to estimate disease severity. The best-performing model, EVA-02-B, achieved strong cross year generalization (R2 = 0.697). Error analysis found that performance was more strongly associated with seasonal disease progression and flight-score time offset than with image-level noise. UAV-based deep learning estimated SLB severity with comparable precision to expert raters. This study lays the groundwork for integrating automated phenotypes into genetic studies of disease resistance. PLAIN LANGUAGE SUMMARYSouthern leaf blight (SLB) of maize is a disease that causes yield loss worldwide and developing resistant varieties offers the best hope for controlling the disease. Studying SLB resistance requires plant pathologists to visually score severity in the field, a labor-intensive method that requires expertise. To address these challenges, we asked whether SLB severity scoring could be automated using drone images and artificial intelligence (AI). We trained AI models using three years of image and score data then compared the results to visual scores taken by five plant pathologists. The best performing AI model showed a similar level of consistency to the experts and proved capable of scoring severity despite unpredictable and uncontrollable conditions that affect field imaging experiments such as weeds or shadows. These findings provide a validated method that improves the efficiency of maize disease research, a critical area of study for agricultural sustainability and productivity.

plant biology↗

bioRxiv2025

Investigating GERMs: How Genotype, Environment, and Rhizosphere Microbiome interactions underlie heat response in maize and sorghum

Plant resistance to heat stress can be modelled by variation attributable to the genotype, environment, the rhizosphere microbiome, and their interactions. Using this Genotype x Environment x Rhizosphere Microbiome (GERMs) model, we studied three cereal genotypes: two inbred maize lines with contrasting heat sensitivity, and a sorghum inbred that displayed moderate heat tolerance. Plants were grown under optimal and heat stressed conditions across two soil treatments. We developed a systems-level metatranscriptomics approach to examine both plant and microbial transcriptomic profiles and integrated them with microbiome compositional data and plant phenotypes. We compared our strategy to amplicon profiling and found that our metatranscriptomic strategy offers greater functional and taxonomic resolution, allowing us to characterize active microbial pathways and analyze them jointly with plant gene expression profiles within a single system. We show that the microbiome functional profile is driven by host genotype and environmental factors and can enhance plant resilience. Our analyses identified plant genes and microbial pathways consistently associated with heat tolerance and key host-microbe interactions. Specifically, we identified D-amino acid metabolism as a plausible mechanism underlying a synergistic response to heat stress. These results demonstrate that the rhizosphere microbiome is not a passive component but an active participant in plant responses to abiotic stress. This work offers a new perspective on cereal adaptation to high temperatures and underscores the utility of the GERMs framework for dissecting functional relationships among plant genotype, environment, and the rhizosphere microbiome.

microbiology↗

bioRxiv2024

Maize inbreds show allelic variation for diel transcription patterns

Circadian entrainment and external cues can cause gene transcript abundance to oscillate throughout the day, and these patterns of diel transcript oscillation vary across genes and plant species. Less is known about within-species allelic variation for diel patterns of transcript oscillation, or about how regulatory sequence variation influences diel transcription patterns. In this study, we evaluated diel transcript abundance for 24 diverse maize inbred lines. We observed extensive natural variation in diel transcription patterns, with two-fold variation in the number of genes that oscillate over the course of the day. A convolutional neural network trained to predict oscillation from promoter sequence identified sequences previously reported as binding motifs for known circadian clock genes in other plant systems. Genes showing diel transcription patterns that cosegregate with promoter sequence haplotypes are enriched for associations with photoperiod sensitivity and may have been indirect targets of selection as maize was adapted to longer day lengths at higher latitudes. These findings support the idea that cis-regulatory sequence variation influences patterns of gene expression, which in turn can have effects on phenotypic plasticity and local adaptation.

plant biology↗

bioRxiv2024

Global Genotype by Environment Prediction Competition Reveals That Diverse Modeling Strategies Can Deliver Satisfactory Maize Yield Estimates

Predicting phenotypes from a combination of genetic and environmental factors is a grand challenge of modern biology. Slight improvements in this area have the potential to save lives, improve food and fuel security, permit better care of the planet, and create other positive outcomes. In 2022 and 2023 the first open-to-the-public Genomes to Fields (G2F) initiative Genotype by Environment (GxE) prediction competition was held using a large dataset including genomic variation, phenotype and weather measurements and field management notes, gathered by the project over nine years. The competition attracted registrants from around the world with representation from academic, government, industry, and non-profit institutions as well as unaffiliated. These participants came from diverse disciplines include plant science, animal science, breeding, statistics, computational biology and others. Some participants had no formal genetics or plant-related training, and some were just beginning their graduate education. The teams applied varied methods and strategies, providing a wealth of modeling knowledge based on a common dataset. The winners strategy involved two models combining machine learning and traditional breeding tools: one model emphasized environment using features extracted by Random Forest, Ridge Regression and Least-squares, and one focused on genetics. Other high-performing teams methods included quantitative genetics, classical machine learning/deep learning, mechanistic models, and model ensembles. The dataset factors used, such as genetics; weather; and management data, were also diverse, demonstrating that no single model or strategy is far superior to all others within the context of this competition.

genetics↗

bioRxiv2022

Yield Prediction Through Integration of Genetic, Environment, and Management Data Through Deep Learning

Accurate prediction of the phenotypic outcomes produced by different combinations of genotypes, environments, and management interventions remains a key goal in biology with direct applications to agriculture, research, and conservation. The past decades have seen an expansion of new methods applied towards this goal. Here we predict maize yield using deep neural networks, compare the efficacy of two model development methods, and contextualize model performance using linear models, which are the conventional method for this task, and machine learning models We examine the usefulness of incorporating interactions between disparate data types. We find a deep learning model with interactions has the best average performance. Optimizing submodules for each datatype improved model performance relative to optimizing the whole model for all data types at once. Examining the effect of interactions in the best performing model revealed that including interactions altered the models sensitivity to weather and management features, including a reduction of the importance scores for timepoints expected to have limited physiological basis for influencing yield - those at the extreme end of the season, nearly 200 days post planting. Based on these results, deep learning provides a promising avenue for phenotypic prediction of complex traits in complex environments and a potential mechanism to better understand the influence of environmental and genetic factors.

plant biology↗

bioRxiv2021

Variation in upstream open reading frames contributes to allelic diversity in protein abundance

The 5 untranslated region (UTR) sequence of eukaryotic mRNAs may contain upstream open reading frames (uORFs), which can regulate translation of the main open reading frame (mORF). The current model of translational regulation by uORFs posits that when a ribosome scans an mRNA and encounters a uORF, translation of that uORF can prevent ribosomes from reaching the mORF and cause decreased mORF translation. In this study, we first observed that rare variants in the 5 UTR dysregulate protein abundance. Upon further investigation, we found that rare variants near the start codon of uORFs can repress or derepress mORF translation, causing allelic changes in protein abundance. This finding holds for common variants as well, and common variants that modify uORF start codons also contribute disproportionately to metabolic and whole-plant phenotypes, suggesting that translational regulation by uORFs serves an adaptive function. These results provide evidence for the mechanisms by which natural sequence variation modulates gene expression, and ultimately, phenotype.

genomics↗

bioRxiv2020

A Maize Practical Haplotype Graph Leverages Diverse NAM Assemblies

As a result of millions of years of transposon activity, multiple rounds of ancient polyploidization, and large populations that preserve diversity, maize has an extremely structurally diverse genome, evidenced by high-quality genome assemblies that capture substantial levels of both tropical and temperate diversity. We generated a pangenome representation (the Practical Haplotype Graph, PHG) of these assemblies in a database, representing the pangenome haplotype diversity and providing an initial estimate of structural diversity. We leveraged the pangenome to accurately impute haplotypes and genotypes of taxa using various kinds of sequence data, ranging from WGS to extremely-low coverage GBS. We imputed the genotypes of the recombinant inbred lines of the NAM population with over 99% mean accuracy, while unrelated germplasm attained a mean imputation accuracy of 92 or 95% when using GBS or WGS data, respectively. Most of the imputation errors occur in haplotypes within European or tropical germplasm, which have yet to be represented in the maize PHG database. Also, the PHG stores the imputation data in a 30,000-fold more space-efficient manner than a standard genotype file, which is a key improvement when dealing with large scale data.

genomics↗

bioRxiv2019

In-field whole plant maize architecture characterized by Latent Space Phenotyping

Collecting useful, interpretable, and biologically relevant phenotypes in a resource-efficient manner is a bottleneck to plant breeding, genetic mapping, and genomic prediction. Autonomous and affordable sub-canopy rovers are an efficient and scalable way to generate sensor-based datasets of in-field crop plants. Rovers equipped with light detection and ranging (LiDar) can produce three-dimensional reconstructions of entire hybrid maize fields. In this study, we collected 2,103 LiDar scans of hybrid maize field plots and extracted phenotypic data from them by Latent Space Phenotyping (LSP). We performed LSP by two methods, principal component analysis (PCA) and a convolutional autoencoder, to extract meaningful, quantitative Latent Space Phenotypes (LSPs) describing whole-plant architecture and biomass distribution. The LSPs had heritabilities of up to 0.44, similar to some manually measured traits, indicating they can be selected on or genetically mapped. Manually measured traits can be successfully predicted by using LSPs as explanatory variables in partial least squares regression, indicating the LSPs contain biologically relevant information about plant architecture. These techniques can be used to assess crop architecture at a reduced cost and in an automated fashion for breeding, research, or extension purposes, as well as to create or inform crop growth models.

plant biology↗

bioRxiv2018

Comparing genome-wide association study results from different measurements of an underlying phenotype

Increasing popularity of high-throughput phenotyping technologies, such as image-based phenotyping, offer novel ways for quantifying plant growth and morphology. These new methods can be more or less accurate and precise than traditional, manual measurements. Many large-scale phenotyping efforts are conducted to enable genome-wide association studies (GWAS), but it is unclear exactly how alternative methods of phenotyping will affect GWAS results. In this study we simulate phenotypes that are controlled by the same set of causal loci but have differing heritability, similar to two different measurements of the same morphological character. We then perform GWAS with the simulated traits and create receiver operating characteristic (ROC) curves from the results. The areas under the ROC curves (AUCs) provide a metric that allows direct comparisons of GWAS results from different simulated traits. We use this framework to evaluate the effects of heritability and the number of causative loci on the AUCs of simulated traits; we also test the differences between AUCs of traits with differing heritability. We find that both increasing the number of causative loci and decreasing the heritability reduce a traits AUC. We also find that when two traits are controlled by a greater number of causative loci, they are more likely to have significantly different AUCs as the difference between their heritabilities increases. These results provide a framework for deciding between competing phenotyping strategies when the ultimate goal is to generate and use phenotype-genotype associations from GWAS.

genomics↗

bioRxiv2018

Selection signatures underlying dramatic male inflorescence transformation during modern hybrid maize breeding

Inflorescence capacity plays a crucial role in reproductive fitness in plants, and in production of hybrid crops. Maize is a monoecious species bearing separate male and female flowers (tassel and ear, respectively). The switch from open-pollinated populations of maize to hybrid-based breeding schemes in the early 20th century was accompanied by a dramatic reduction in tassel size, and the trend has continued with modern breeding over the recent decades. The goal of this study was to identify selection signatures in genes that may underlie this dramatic transformation. Using a population of 942 diverse inbred maize accessions and a nested association mapping population comprised of three 200-line biparental populations, we measured 15 tassel morphological characteristics by manual and image-based methods. Genome-wide association studies identified 242 single nucleotide polymorphisms significantly associated with measured traits. We compared 41 unselected lines from the Iowa Stiff Stalk Synthetic (BSSS) population to 21 highly selected lines developed by modern commercial breeding programs and show that tassel size and weight were reduced significantly. We assayed genetic differences between the two groups using selection statistics XP-EHH, XP-CLR, and FST. All three selection statistics show evidence of selection at genomic regions associated with tassel morphology relative to genome-wide null distributions. These results support the tremendous effect, both phenotypic and genotypic, that selection has had on maize male inflorescence morphology.

genetics↗