Biology subjects

Wallace, J. G.

Publications and source records attributed to Wallace, J. G..

9 recordsLinked to original sources

bioRxiv2026

Rhizobacterial Biosensors Spatially Map Natural and Engineered Sucrose Exudation

Root exudation mediates the delivery of plant primary and secondary metabolites into soil, where they regulate plant-microbe interactions and terrestrial carbon cycling. Conventional exudate analyses quantify total root-released carbon yet obscure the spatial origin and rhizosphere influence of individual compounds. Here, we develop a rhizobacterial biosensor platform, named Suc-MAPP, to map local exudate profiles along the surface of colonized root tissues. Focusing on sucrose, we engineered sfGFP-based, sucrose-responsive gene circuits in Pseudomonas putida KT2440 for live imaging of exudate concentrations in the micromolar range. These biosensors reveal spatially structured sucrose exudation patterns across eudicots and monocots and implicate photoassimilated source-sink dynamics as a major determinant. We further apply this platform to phenotype exudation modulated by synthetic gene circuitry in Arabidopsis thaliana, identifying genetic design rules for graded sucrose release and quantifying how engineered export sculpts rhizosphere assembly of a defined bacterial community. Together, these results establish programmable rhizobacterial biosensors as tools to spatially resolve plant-environment carbon exchange in situ and provide a framework for extending this approach to diverse exudate targets.

synthetic biology↗

bioRxiv2025

Gene-by-Environment Interaction Significantly Drives Bacterial Endophyte Communities in Maize Stalks

1.BackgroundMany microbes are known to boost the performance of crop plants, and their use as crop treatments ("biologicals") is an active area of research and commercialization. Although many identified microbes can boost plant production under laboratory settings, most fail to translate to field settings, and even the ones that are commercialized are often unreliable. While many factors likely underlie these issues, one that has been underexplored is the idea that interactions between plants and microbes can vary across both different host genetics and different environments, and that specific combinations of host and environment can result in very different microbial outcomes. ResultsTo quantify how a plant microbiome can change across host genotype and environment and to determine the relative importance of these factors, we sampled maize stalks from 20 specific hybrid varieties grown at 15 locations in the United States. Using 16S rRNA gene sequencing, we find that the stalk microbiome is highly variable, with a handful of bacterial taxa conserved at broader taxonomic levels (Phylum, Class) and almost none at finer levels (Genus, Species). Local environment and gene-by-environment interactions both had consistently large and significant effects on the microbial community (measured by alpha-and beta-diversity summary statistics, individual taxa, and inferred functional capacity), whereas host genotype had little to no consistent effect across environments. We also identified specific soil factors (pH and potassium) as the most significant environmental drivers of the community, even though these communities were living inside the stalks. ConclusionsTaken together, our results indicate that while host genetics can significantly affect the stalk microbial community, these effects are almost entirely predicated on interactions with the environment. These results imply that reliable deployment of microbes for crop production will require significant investment in trials across many locations and genetic backgrounds, either to estimate the GxE effects directly or to screen for microbes whose effects are less dependent on these two factors.

microbiology↗

bioRxiv2025

Genetic determinants of aerial root traits that support biological nitrogen fixation in maize

Modern agriculture depends on chemically synthesized nitrogen fertilizer, which ensures high yields but also can carry significant environmental and economic costs. Biological nitrogen fixation (BNF) already supplies nitrogen to legume crops and several avenues of research are underway to extend it to non-legume crops. In maize (Zea mays), aerial roots have been shown to contribute to BNF in some varieties, and both having many aerial roots and large aerial roots contributes to the fixation trait. However, much of the genetics controlling aerial root number and size is still unknown. Here we validate and quantify BNF in maize varieties from Southern Mexico under controlled conditions and evaluate a population of double haploids derived from the elite inbred PHZ51 crossed with these varieties. We find that most aerial root traits (root number, nodes with roots, root size) are reasonably heritable (h2 0.5-0.75) and generally uncorrelated with each other. QTL mapping identifies 5 QTL each affecting nodes with aerial roots and aerial root number per node; in both cases all but 1 QTL show an increase from the landrace allele. We also identify 11 QTL for aerial root diameter, with most positive QTL coming from PHZ51. Between the two populations, only a few QTL overlap, indicating a presumably high diversity of genes affecting aerial root morphology in landrace populations. Combining the best QTL into elite material may provide a path toward meaningful levels of BNF for maize, and additional work is needed to determine how viable this approach will be in field settings.

plant biology↗

bioRxiv2023

kGWASflow: a modular, flexible, and reproducible Snakemake workflow for k-mers-based GWAS

Genome-wide association studies (GWAS) have been widely used to identify genetic variation associated with complex traits. Despite its success and popularity, the traditional GWAS approach comes with a variety of limitations. For this reason, newer methods for GWAS have been developed, including the use of pan-genomes instead of a reference genome and the utilization of markers beyond single-nucleotide polymorphisms, such as structural variations and k-mers. The k-mers based GWAS approach has especially gained attention from researchers in recent years. However, these new methodologies can be complicated and challenging to implement. Here we present kGWASflow, a modular, user-friendly, and scalable workflow to perform GWAS using k-mers. We adopted an existing kmersGWAS method into an easier and more accessible workflow using management tools like Snakemake and Conda and eliminated the challenges caused by missing dependencies and version conflicts. kGWASflow increases the reproducibility of the kmersGWAS method by automating each step with Snakemake and using containerization tools like Docker. The workflow encompasses supplemental components such as quality control, read-trimming procedures, and generating summary statistics. kGWASflow also offers post-GWAS analysis options to identify the genomic location and context of trait-associated k-mers. kGWASflow can be applied to any organism and requires minimal programming skills. kGWASflow is freely available on GitHub (https://github.com/akcorut/kGWASflow) and Bioconda (https://anaconda.org/bioconda/kgwasflow).

bioinformatics↗

bioRxiv2021

Gut microbiota-based vaccination engages innate immunity to improve blood glucose control in obese mice

Obesity and diabetes increase circulating levels of microbial components derived from the gut microbiota. Individual bacterial factors (i.e., postbiotics) can have opposing effects on metabolic inflammation and blood glucose control. We tested the net effect of gut bacterial extracts on blood glucose using a microbiota-based vaccination strategy in mice. Male and female mice had improved insulin sensitivity and blood glucose control five weeks after a single subcutaneous injection of a specific dose of a bacterial extract obtained from the luminal contents of the proximal gut. Injection of mice with proximal gut extracts from germ-free mice revealed that bacteria were required for a microbiota-based vaccination to improve blood glucose control. Vaccination of Nod1-/-, Nod2-/-, and Ripk2-/- mice showed that each of these innate immune proteins was required for bacterial extract injection to improve blood glucose control. A microbiota-based vaccination promoted a proximal gut immunoglobulin-G (IgG) response directed against bacterial extract antigens, where subcutaneous injection of mice with the luminal contents of the ileum elicited a bacterial extract-specific IgG response that is compartmentalized to the ileum of vaccinated mice. A microbiota-based vaccination was associated with an altered the microbiota composition in the ileum and colon of mice. Lean mice required a single injection of proximal gut bacterial extracts, but high fat diet (HFD)-fed, obese mice required prime-boost bacterial extract injections for improvements in blood glucose control. These data show that, upon subversion of the gut barrier, vaccination with proximal gut bacterial extracts engages innate immunity to promote long-lasting improvements in blood glucose control in a dose-dependent manner.

physiology↗

bioRxiv2021

Genomic and Chemical Diversity of Commercially Available Industrial Hemp Accessions

High consumer demand for cannabidiol (CBD) has made industrial hemp (Cannabis sativa) an extremely high-value crop. However, high demand has resulted in the industry developing faster than the research, resulting in the sale of many hemp accessions with inconsistent performance and chemical profiles. To determine the genetic and phenotypic consistency in available CBD hemp varieties, we obtained seed or clones from 22 different named accessions. Genotypes (~48,000 SNPs) and chemical profiles (% CBD and THC by dry weight) were determined for up to 8 plants per accession. Many accessions--including several with the same name--showed little consistency either genetically or chemically. Most seed-grown accessions deviated significantly from their purported levels of CBD and THC based on the supplied certificates of analysis. Several also showed evidence of an active tetrahydrocannabinolic acid (THCa) synthase gene, leading to unacceptably high levels of THC in female flowers. We conclude that the current market for CBD-hemp varieties is highly unreliable, making many purchases risky for growers. We suggest options for addressing these issues, such using unique names and developing seed and plant certification programs to ensure the availability of high-quality, verified planting materials.

genomics↗

bioRxiv2020

Eleven biosynthetic genes explain the majority of natural variation for carotenoid levels in maize grain

Vitamin A deficiency remains prevalent in parts of Asia, Latin America, and sub-Saharan Africa where maize is a food staple. Extensive natural variation exists for carotenoids in maize grain; to understand its genetic basis, we conducted a joint linkage and genome-wide association study in the U.S. maize nested association mapping panel. Eleven of the 44 detected quantitative trait loci (QTL) were resolved to individual genes. Six of these were correlated expression and effect QTL (ceeQTL), showing strong correlations between RNA-seq expression abundances and QTL allelic effect estimates across six stages of grain development. These six ceeQTL also had the largest percent phenotypic variance explained, and in major part comprised the three to five loci capturing the bulk of genetic variation for each trait. Most of these ceeQTL had strongly correlated QTL allelic effect estimates across multiple traits. These findings provide the most comprehensive genome-level understanding of the genetic and molecular control of carotenoids in any plant system, and a roadmap to accelerate breeding for provitamin A and other priority carotenoid traits in maize grain that should be readily extendable to other cereals.

genetics↗

bioRxiv2018

High-fat diet intake modulates maternal intestinal adaptations to pregnancy, and results in placental hypoxia and impaired fetal gut development

Shifts in maternal intestinal microbiota have been implicated in metabolic adaptations to pregnancy. In this study we investigated how high-fat diet intake impacts the maternal gut microbiota, intestinal inflammation and gut barrier integrity, placental inflammation, and fetal intestinal development at E18.5. High-fat diet (HFD) was associated with decreased relative abundancesof SCFA producing genera during pregnancy. These diet-induced shifts paralleled decreased maternal intestinal mRNA levels of SCFA receptor Gpr41, modestly decreased cecal butyrate, and altered mRNA levels of inflammatory cytokines and immune cell markers in the maternal intestine. Maternal HFD resulted inimpaired gut barrier integrity, with corresponding increases in circulating maternal levels of LPS and TNF.Placentafromhigh-fat fed damsdemonstrated blood vessel immatu-rityand hypoxia, decreased freecarnitine, acylcarnitine derivatives, TMAO, as well as altered mRNA levels of inflammation, autophagy and ER stress markers. HFD exposed fetuses had increased activation of NF-{kappa}B and inhibition of the unfolded protein response in the developing intestine. Together, these data suggest that high-fat diet intake prior to and during pregnancy shifts the composition of the maternal gut microbiota and impairs gut barrier integrity, resulting in increased maternal circulating LPS, which may ultimate contribute to changes in placental vasculariza-tion and fetal gut development.\n\nFunding informationFarncombe Family Digestive Health Research Institute (KMK); Canadian Institute of Health Research (CJB); Canada Research Chairs Program (MGS, DMS); Natural Sciences and Engineering Research Council of Canada, Genome Canada (PBM).

developmental biology↗

bioRxiv2018

Quantitative Genetic Analysis of the Maize Leaf Microbiome

The degree to which an organism can affect its associated microbial communities (\"microbiome\") varies by organism and habitat, and in many cases is unknown. We address this question by analyzing the metabolically active bacteria of the maize phyllosphere across 300 diverse maize lines growing in a common environment. We performed comprehensive heritability analysis for 49 community diversity metrics, 380 bacterial clades (individual operational taxonomic units and higher-level groupings), and 9042 predicted metagenomic functions. We find that only a few few bacterial clades (5) and diversity metrics (2) are significantly heritable, while a much larger number of metabolic functions (200) are. Many of these associations appear to be driven by the amount of Methylobacteria present in each sample, and we find significant enrichment for traits relating to short-chain carbon metabolism, secretion, and nitrotoluene degradation. Genome-wide association analysis identifies a small number of associated loci for these heritable traits, including two loci (on maize chromosomes 7 and 10) that affect a large number of traits even after correcting for correlations among traits. This work is among the most comprehensive analyses of the maize phyllosphere to date. Our results indicate that while most of the maize phyllosphere composition is driven by environmental factors and/or stochastic founder events, a subset of bacterial taxa and metabolic functions is nonetheless significantly impacted by host plant genetics. Additional work will be needed to identify the exact nature of these interactions and what effects they may have on the phenotype of host plants.

genetics↗