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Andorf, C. M.

Publications and source records attributed to Andorf, C. M..

14 recordsLinked to original sources

Visualizing and integrating linear and graph pangenomes at the Maize Genetics and Genomics Database

Genome visualization has existed for as long as assembled genomes, with an array of tools and approaches to meet researchers' needs. The genome browser is one of these tools, and it has become a critical analytical resource for geneticists, genome biologists, and breeders. The evolution of genome browser capabilities is ongoing, and the USDA-ARS Maize Genetics and Genomics Database (MaizeGDB) has implemented JBrowse2, the most recent iteration of JBrowse. It includes multiple genome browser and alignment views, demonstrating pangenome visualization capability and permitting sophisticated functional characterization of maize loci. Described here also is MaizeGDB's new Pangenome Viewer, which integrates pangenome graph views with linear browser visualization to obtain an on-the-fly, interactive visual snapshot of structural variation across a pangenome at user-selected loci, with zoom capabilities and statistical and variant information for each subgraph. Finally, we demonstrate how different MaizeGDB pangenome pipelines complement one another to help guide accurate analyses. This functionality can lead to more precise characterization of loci that confer important agronomic traits, resulting in better outcomes for farmers and the public.

genomics↗

Why do some predicted protein structures fold poorly? Benchmarking AlphaFold, ESMFold, and Boltz in maize

Protein structure prediction tools have significantly reduced the time and cost to generate protein structures and accelerated protein discovery and design. However, plant proteins are underrepresented in sequence and structural datasets used to train these programs. To quantify the downstream impact of this deficiency, we benchmarked five structure-prediction programs (AlphaFold 2, AlphaFold 3, ESMFold, Boltz-1, and Boltz-2) across 417 well-characterized Zea mays genes. These "classical" genes represent a set of well-studied genes with known genetic and phenotypic effects. We generated structures for each gene using these programs and compared how sequence, structural, and evolutionary conservation impacted the structures confidence and geometric features. Proteins lacking conserved sequence and/or structural domains had on average 25% to 43% lower confidence scores than proteins having both domains. Proteome-wide phylostratigraphy revealed that species-specific proteins had substantially lower confidence scores than proteins conserved amongst angiosperms and Eukaryotes. Boltz-1 and ESMFold structures had the highest occurrence of structures with severe geometry issues, including overlapping atoms and unlikely bond angles. We also compared computational and experimental alignments of Arabidopsis, maize, rice, wheat, and soybean proteins from the Protein Data Bank and identified structures showing incongruences with experimental data. This study challenges the assertion that protein folding has been completely solved, and urges more investigation into benchmarking and standardized evaluation frameworks to improve model performance and assessment in agricultural crops.

bioinformatics↗

Extensive genome evolution distinguishes maize within a stable tribe of grasses

Over the last 20 million years, the Andropogoneae tribe of grasses has evolved to dominate 17% of global land area. Domestication of these grasses in the last 10,000 years has yielded our most productive crops, including maize, sugarcane, and sorghum. The majority of Andropogoneae species, including maize, show a history of polyploidy - a condition that, while offering the evolutionary advantage of multiple gene copies, poses challenges to basic cellular processes, gene expression, and epigenetic regulation. Genomic studies of polyploidy have been limited by sparse sampling of taxa in groups with multiple polyploidy events. Here, we present 33 genome assemblies from 27 species, including chromosome-scale assemblies of maize relatives Zea and Tripsacum. In maize, the after-effects of polyploidy have been widely studied, showing reduced chromosome number, biased fractionation of duplicate genes, and transposable element (TE) expansions. While we observe these patterns within the genus Zea, 12 other polyploidy events deviate significantly. Those tetraploids and hexaploids retain elevated chromosome number, maintain nearly complete complements of duplicate genes, and have only stochastic TE amplifications. These genomes reveal variable outcomes of polyploidy, challenging simple predictions and providing a foundation for understanding its evolutionary implications in an ecologically and economically important clade.

evolutionary biology↗

Application of RFdiffusion to predict interspecies protein-protein interactions between fungal pathogens and cereal crops

Plant pathogenic fungi secrete small proteins known as effectors which help overcome the plant defense response and cause disease. The concept of effector-triggered immunity in plants evolved from the "gene for gene hypothesis" which describes plant resistance or susceptibility to plant pathogens based on interspecies protein-protein interactions (PPIs) between plant-derived resistance (R) genes and pathogen-derived avirulence (Avr) effector genes. Understanding the molecular interactions mediating these host-pathogen interactions in effector-triggered immunity is thus essential to managing fungal disease. In silico methods of predicting interspecies PPIs have been heavily studied to identify target genes for crop resistance. But conventional sequence-based homology methods (i.e., interlog, domain-based inference) for predicting interspecies PPIs are not as powerful as methods that also incorporate structural homology. The objective of this study was to develop a computational workflow to predict PPIs between pathogenic fungi and their cereal hosts by leveraging recent advances in artificial intelligence and structural biology. This workflow proposes the use of a generative model, RFdiffusion, to predict the structure of truncated segments of proteins likely to bind to query effector proteins. The binder structures were filtered based on the number of contacts at the effectors known binding residues. Acceptable structures were then input into FoldSeek to search the host proteome for host proteins containing similar sub-structures. Experimentally-validated PPIs between rice (Oryzae sativa cv. Japonica) and rice blast fungus (Magnaporthe oryzae) were used for workflow validation. The effects of binder length and the binding residues mode of action (i.e., residues at active/substrate recognition sites) on the binder quality and presumptive host protein matches were explored. Ultimately, 11 out of 14 experimentally validated PPIs were recovered computationally, indicating a high recall (>78%) for the workflow. The shorter binders recovered most of the PPIs, but may have produced the most false positives, as functional analyses revealed that these host proteins displayed a wide variety of functions. These findings emphasize that subject matter expertise is still required to decipher the prediction results. Yet, this framework for elucidating interactions between fungal pathogens and host proteins could provide valuable insight into mechanisms of susceptibility or resistance at a scale friendly to limited computational resources, and facilitate the development of control strategies that reduce crop diseases.

plant biology↗

Fusarium Protein Toolkit: AI-powered tools to combat fungal threats to agriculture

BackgroundThe fungal genus Fusarium poses significant threats to food security and safety worldwide because it consists of numerous species that cause destructive diseases in crops, as well as mycotoxin contamination. The adverse effects of climate change are exacerbating some existing threats and causing new problems. These challenges highlight the need for innovative solutions, including the development of advanced tools to identify targets to control crop diseases and mycotoxin contamination incited by Fusarium. DescriptionIn response to these challenges, we developed the Fusarium Protein Toolkit (FPT, https://fusarium.maizegdb.org/), a web-based tool that allows users to interrogate the structural and variant landscape within the Fusarium pan-genome. FPT offers a comprehensive approach to understanding and mitigating the detrimental effects of Fusarium on agriculture. The tool displays both AlphaFold and ESMFold-generated protein structure models from six Fusarium species. The structures are accessible through a user-friendly web portal and facilitate comparative analysis, functional annotation inference, and identification of related protein structures. Using a protein language model, FPT predicts the impact of over 270 million coding variants in two of the most agriculturally important species, Fusarium graminearum, which causes Fusarium head blight and trichothecene mycotoxin contamination of cereals, and F. verticillioides, which causes ear rot and fumonisin mycotoxin contamination of maize. To facilitate the assessment of naturally occurring genetic variation, FPT provides variant effect scores for proteins in a Fusarium pan-genome constructed from 22 diverse species. The scores indicate potential functional consequences of amino acid substitutions and are displayed as intuitive heatmaps using the PanEffect framework. ConclusionFPT fills a knowledge gap by providing previously unavailable tools to assess structural and missense variation in proteins produced by Fusarium, the most agriculturally important group of mycotoxin-producing plant pathogens. FPT will deepen our understanding of pathogenic mechanisms in Fusarium, and aid the identification of genetic targets that can be used to develop control strategies that reduce crop diseases and mycotoxin contamination. Such targets are vital to solving the agricultural problems incited by Fusarium, particularly evolving threats affected by climate change. By providing a novel approach to interrogate Fusarium-induced crop diseases, FPT is a crucial step toward safeguarding food security and safety worldwide.

bioinformatics↗

A unified VCF data set from nearly 1,500 diverse maize accessions and resources to explore the genomic landscape of maize

Efforts to capture and analyze maize nucleotide diversity have ranged widely in scope, but differences in reference genome version and software algorithms used in these efforts inhibit comparison. To address these continuity issues, The Maize Genetics and Genomics Database has collaborated with researchers in the maize community to offer variant data from a diverse set of 1,498 inbred lines, traditional varieties, and teosintes through a standardized variant-calling pipeline against version 5 of the B73 reference genome. The output was filtered for mapping quality, coverage, and linkage disequilibrium, and annotated based on variant effects relative to the B73 RefGen_v5 gene annotations. MaizeGDB has also updated a web tool to filter, visualize, and download genotype sets based on genomic locations and accessions of interest. MaizeGDB plans to host regular updates of these resources as additional resequencing data become available, with plans to expand to all publicly available sequence data.

genomics↗

PanEffect: A pan-genome visualization tool for variant effects in maize

Understanding the effects of genetic variants is crucial for accurately predicting traits and phenotypic outcomes. Recent advances have utilized protein language models to score all possible missense variant effects at the proteome level for a single genome, but a reliable tool is needed to explore these effects at the pan-genome level. To address this gap, we introduce a new tool called PanEffect. We implemented PanEffect at MaizeGDB to enable a comprehensive examination of the potential effects of coding variants across 51 maize genomes. The tool allows users to visualize over 550 million possible amino acid substitutions in the B73 maize reference genome and also to observe the effects of the 2.3 million natural variations in the maize pan-genome. Each variant effect score, calculated from the Evolutionary Scale Modeling (ESM) protein language model, shows the log-likelihood ratio difference between B73 and all variants in the pan-genome. These scores are shown using heatmaps spanning benign outcomes to strong phenotypic consequences. Additionally, PanEffect displays secondary structures and functional domains along with the variant effects, offering additional functional and structural context. Using PanEffect, researchers now have a platform to explore protein variants and identify genetic targets for crop enhancement. Availability and implementation: The PanEffect code is freely available on GitHub (https://github.com/Maize-Genetics-and-Genomics-Database/PanEffect). A maize implementation of PanEffect and underlying datasets are available at MaizeGDB (https://www.maizegdb.org/effect/maize/).

bioinformatics↗

FASSO: An AlphaFold based method to assign functional annotations by combining sequence and structure orthology

Methods to predict orthology play an important role in bioinformatics for phylogenetic analysis by identifying orthologs within or across any level of biological classification. Sequence-based reciprocal best hit approaches are commonly used in functional annotation since orthologous genes are expected to share functions. The process is limited as it relies solely on sequence data and does not consider structural information and its role in function. Previously, determining protein structure was highly time-consuming, inaccurate, and limited to the size of the protein, all of which resulted in a structural biology bottleneck. With the release of AlphaFold, there are now over 200 million predicted protein structures, including full proteomes for dozens of key organisms. The reciprocal best structural hit approach uses protein structure alignments to identify structural orthologs. We propose combining both sequence- and structure-based reciprocal best hit approaches to obtain a more accurate and complete set of orthologs across diverse species, called Functional Annotations using Sequence and Structure Orthology (FASSO). Using FASSO, we annotated orthologs between five plant species (maize, sorghum, rice, soybean, Arabidopsis) and three distance outgroups (human, budding yeast, and fission yeast). We inferred over 270,000 functional annotations across the eight proteomes including annotations for over 5,600 uncharacterized proteins. FASSO provides confidence labels on ortholog predictions and flags potential misannotations in existing proteomes. We further demonstrate the utility of the approach by exploring the annotation of the maize proteome.

bioinformatics↗

Association Mapping Across a Multitude of Traits Collected in Diverse Environments Identifies Pleiotropic Loci in Maize.

Classical genetic studies have identified many cases of pleiotropy where mutations in individual genes alter many different phenotypes. Quantitative genetic studies of natural genetic variants frequently examine one or a few traits, limiting their potential to identify pleiotropic effects of natural genetic variants. Widely adopted community association panels have been employed by plant genetics communities to study the genetic basis of naturally occurring phenotypic variation in a wide range of traits. High-density genetic marker data - 18M markers - from two partially overlapping maize association panels comprising 1,014 unique genotypes grown in field trials across at least seven US states and scored for 162 distinct trait datasets enabled the identification of of 2,154 suggestive marker-trait associations and 697 confident associations in the maize genome using a resampling-based genome-wide association strategy. The precision of individual marker-trait associations was estimated to be three genes based a reference set of genes with known phenotypes. Examples were observed of both genetic loci associated with variation in diverse traits (e.g. above-ground and below-ground traits), as well as individual loci associated with the same or similar traits across diverse environments. Many significant signals are located near genes whose functions were previously entirely unknown or estimated purely via functional data on homologs. This study demonstrates the potential of mining community association panel data using new higher density genetic marker sets combined with resampling-based genome-wide association tests to develop testable hypotheses about gene functions, identify potential pleiotropic effects of natural genetic variants, and study genotype by environment interaction.

plant biology↗

FINDER: An automated software package to annotate eukaryotic genes from RNA-Seq data and associated protein sequences

BackgroundGene annotation in eukaryotes is a non-trivial task that requires meticulous analysis of accumulated transcript data. Challenges include transcriptionally active regions of the genome that contain overlapping genes, genes that produce numerous transcripts, transposable elements and numerous diverse sequence repeats. Currently available gene annotation software applications depend on pre-constructed full-length gene sequence assemblies which are not guaranteed to be error-free. The origins of these sequences are often uncertain, making it difficult to identify and rectify errors in them. This hinders the creation of an accurate and holistic representation of the transcriptomic landscape across multiple tissue types and experimental conditions. Therefore, to gauge the extent of diversity in gene structures, a comprehensive analysis of genome-wide expression data is imperative. ResultsWe present FINDER, a fully automated computational tool that optimizes the entire process of annotating genes and transcript structures. Unlike current state-of-the-art pipelines, FINDER automates the RNA-Seq pre-processing step by working directly with raw sequence reads and optimizes gene prediction from BRAKER2 by supplementing these reads with associated proteins. The FINDER pipeline (1) reports transcripts and recognizes genes that are expressed under specific conditions, (2) generates all possible alternatively spliced transcripts from expressed RNA-Seq data, (3) analyzes read coverage patterns to modify existing transcript models and create new ones, and (4) scores genes as high- or low-confidence based on the available evidence across multiple datasets. We demonstrate the ability of FINDER to automatically annotate a diverse pool of genomes from eight species. ConclusionsFINDER takes a completely automated approach to annotate genes directly from raw expression data. It is capable of processing eukaryotic genomes of all sizes and requires no manual supervision - ideal for bench researchers with limited experience in handling computational tools.

bioinformatics↗

De novo assembly, annotation, and comparative analysis of 26 diverse maize genomes

We report de novo genome assemblies, transcriptomes, annotations, and methylomes for the 26 inbreds that serve as the founders for the maize nested association mapping population. The data indicate that the number of pan-genes exceeds 103,000 and that the ancient tetraploid character of maize continues to degrade by fractionation to the present day. Excellent contiguity over repeat arrays and complete annotation of centromeres further reveal the locations and internal structures of major cytological landmarks. We show that combining structural variation with SNPs can improve the power of quantitative mapping studies. Finally, we document variation at the level of DNA methylation, and demonstrate that unmethylated regions are enriched for cis-regulatory elements that overlap QTL and contribute to changes in gene expression. One sentence summaryA multi-genome analysis of maize reveals previously unknown variation in gene content, genome structure, and methylation.

genomics↗

GenomeQC: A quality assessment tool for genome assemblies and gene structure annotations

BackgroundGenome assemblies are foundational for understanding the biology of a species. They provide a physical framework for mapping additional sequences, thereby enabling characterization of, for example, genomic diversity and differences in gene expression across individuals and tissue types. Quality metrics for genome assemblies gauge both the completeness and contiguity of an assembly and help provide confidence in downstream biological insights. To compare quality across multiple assemblies, a set of common metrics are typically calculated and then compared to one or more gold standard reference genomes. While several tools exist for calculating individual metrics, applications providing comprehensive evaluations of multiple assembly features are, perhaps surprisingly, lacking. Here, we describe a new toolkit that integrates multiple metrics to characterize both assembly and gene annotation quality in a way that enables comparison across multiple assemblies and assembly types.\n\nFindingsOur application, named GenomeQC, is an easy-to-use and interactive web framework that integrates various quantitative measures to characterize genome assemblies and annotations. GenomeQC provides researchers with a comprehensive summary of these statistics and allows for benchmarking against gold standard reference assemblies.\n\nConclusionsThe GenomeQC web application is implemented in R/Shiny version 1.5.9 and Python 3.6 and is freely available at https://genomeqc.maizegdb.org/ under the GPL license.\n\nAll source code and a containerized version of the GenomeQC pipeline is available in the GitHub repository https://github.com/HuffordLab/GenomeQC.

bioinformatics↗

Crowdsourcing Image Analysis for Plant Phenomics to Generate Ground Truth Data for Machine Learning

The accuracy of machine learning tasks critically depends on high quality ground truth data. Therefore, in many cases, producing good ground truth data typically involves trained professionals; however, this can be costly in time, effort, and money. Here we explore the use of crowdsourcing to generate a large number of training data of good quality. We explore an image analysis task involving the segmentation of corn tassels from images taken in a field setting. We investigate the accuracy, speed and other quality metrics when this task is performed by students for academic credit, Amazon MTurk workers, and Master Amazon MTurk workers. We conclude that the Amazon MTurk and Master Mturk workers perform significantly better than the for-credit students, but with no significant difference between the two MTurk worker types. Furthermore, the quality of the segmentation produced by Amazon MTurk workers rivals that of an expert worker. We provide best practices to assess the quality of ground truth data, and to compare data quality produced by different sources. We conclude that properly managed crowdsourcing can be used to establish large volumes of viable ground truth data at a low cost and high quality, especially in the context of high throughput plant phenotyping. We also provide several metrics for assessing the quality of the generated datasets.\n\nAuthor SummaryFood security is a growing global concern. Farmers, plant breeders, and geneticists are hastening to address the challenges presented to agriculture by climate change, dwindling arable land, and population growth. Scientists in the field of plant phenomics are using satellite and drone images to understand how crops respond to a changing environment and to combine genetics and environmental measures to maximize crop growth efficiency. However, the terabytes of image data require new computational methods to extract useful information. Machine learning algorithms are effective in recognizing select parts of images, but they require high quality data curated by people to train them, a process that can be laborious and costly. We examined how well crowdsourcing works in providing training data for plant phenomics, specifically, segmenting a corn tassel - the male flower of the corn plant - from the often-cluttered images of a cornfield. We provided images to students, and to Amazon MTurkers, the latter being an on-demand workforce brokered by Amazon.com and paid on a task-by-task basis. We report on best practices in crowdsourcing image labeling for phenomics, and compare the different groups on measures such as fatigue and accuracy over time. We find that crowdsourcing is a good way of generating quality labeled data, rivaling that of experts.

plant biology↗

Maize GO Annotation - Methods, Evaluation, and Review (maize-GAMER)

1We created a new high-coverage, robust, and reproducible functional annotation of maize protein coding genes based on Gene Ontology (GO) term assignments. Whereas the existing Phytozome and Gramene maize GO annotation sets only cover 41% and 56% of maize protein coding genes, respectively, this study provides annotations for 100% of the genes. We also compared the quality of our newly-derived annotations with the existing Gramene and Phytozome functional annotation sets by comparing all three to a manually annotated gold standard set of 1,619 genes where annotations were primarily inferred from direct assay or mutant phenotype. Evaluations based on the gold standard indicate that our new annotation set is measurably more accurate than those from Phytozome and Gramene. To derive this new high-coverage, high-confidence annotation set we used sequence-similarity and protein-domain-presence methods as well as mixed-method pipelines that developed for the Critical Assessment of Function Annotation (CAFA) challenge. Our project to improve maize annotations is called maize-GAMER (GO Annotation Method, Evaluation, and Review) and the newly-derived annotations are accessible via MaizeGDB (http://download.maizegdb.org/maize-GAMER) and CyVerse (B73 RefGen_v3 5b+ at doi: doi.org/10.7946/P2S62P and B73 RefGen_v4 Zm00001d.2 at doi: doi.org/10.7946/P2M925).

bioinformatics↗