The Maize Genetics and Genomics Database (MaizeGDB) is the community resource for maize researchers, offering a suite of tools, informatics resources, and curated data sets to support maize genetics, genomics, and breeding research. Here, we provide an overview of the key resources available at MaizeGDB, including maize genomes, comparative genomics, and pan-genomics tools. This review aims to familiarize users with the range of options available for maize research and highlights the importance of MaizeGDB as a central hub for the maize research community. By providing a detailed snapshot of the database's capabilities, we hope to enable researchers to make use of MaizeGDB's resources, ultimately assisting them to better study the evolution and diversity of maize.

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.

Pan-genomes, encompassing the entirety of genetic sequences found in a collection of genomes within a clade, are more useful than single reference genomes for studying species diversity. This is especially true for a species like Zea mays, which has a particularly diverse and complex genome. Presenting pan-genome data, analyses, and visualization is challenging, especially for a diverse species, but more so when pan-genomic data is linked to extensive gene model and gene data, including classical gene information, markers, insertions, expression and proteomic data, and protein structures as is the case at MaizeGDB. Here, we describe MaizeGDB's expansion to include the genic subset of the Zea pan-genome in a pan-gene data center featuring the maize genomes hosted at MaizeGDB, and the outgroup teosinte Zea genomes from the Pan-Andropoganeae project. The new data center offers a variety of browsing and visualization tools, including sequence alignment visualization, gene trees and other tools, to explore pan-genes in Zea that were calculated by the pipeline Pandagma. Combined, these data will help maize researchers study the complexity and diversity of Zea, and to use the comparative functions to validate pan-gene relationships for a selected gene model.

Histone deacetylases (HDACs) are important regulators of gene transcription thus controlling multiple cellular processes. Despite its essential role in plants,HDA6is yet to be validated in common bean. In this study, we show thatHDA6is involved in plant development and stress response. Differential expression ofHDA6was determined in various tissues and the expression was seen to be upregulated with plant age (seedling < flowering < maturity). Higher expression was observed in flowers and pods than in stem, leaf, and root. Upregulation ofHDA6gene during cold stress implies its prominent role in abiotic stress. Furthermore, theHDA6gene was isolated from three common bean genotypes and sequence analyses revealed homology with functionally characterized homologs in model species. The 53 kDa translated product was detected using anHDA6specific antibody and recombinant protein overexpressed inEscherichia colishowed HDAC activityin vitro. To our knowledge, this is the first report in the agriculturally important crop common bean describing the functional characterization and biological role ofHDA6.

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.

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/ ).