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Caseys, C.

Publications and source records attributed to Caseys, C..

15 recordsLinked to original sources

A genetic toolkit to reduce wheat immunogenicity and incidence of celiac disease

Celiac disease (CeD) is an immune-mediated condition triggered by wheat gluten in genetically predisposed individuals. The immune reaction in people with CeD is driven by particular gluten amino acid sequences, or immunogenic epitopes. Some of these epitopes elicit strong immune responses in the majority of CeD patients and are designated as immunodominant epitopes. Previous research has shown correlations between the amount of immunogenic wheat epitopes consumed and the onset of CeD, suggesting that reducing wheat immunogenic epitopes may reduce CeD incidence at the population level. Gluten consists of gliadins and glutenins, with gliadins having the majority of the immunodominant epitopes and glutenins playing a major role in dough strength and breadmaking quality (BMQ). This study used radiation-induced deletions, chemical mutagenesis, and natural variation in wheat (Triticum aestivum) to generate genetic stocks with reduced immunogenic epitope content. Most lines were developed in the wheat cultivar Summit, for which we produced a full genome assembly and annotation. We used exome capture to characterize these deletions and identify prolamins located within and outside the deletions. We combined different deletions and developed molecular markers to facilitate their deployment. For chromosome arms 1BS and 1DS, we generated two alternative lines: one lacking immunogenic epitopes for the development of CeD-safe genetic stocks for research purposes, and another retaining selected glutenins for breeding commercial lines with reduced immunogenicity and adequate BMQ. By making these non-transgenic genetic stocks publicly available, we aim to accelerate the development of wheat varieties with reduced immunogenicity and, eventually, a fully CeD-safe wheat.

plant biology↗

A co-proteomic view of metabolite-specific interactions in the Botrytis cinerea-Arabidopsis pathosystem

To successfully infect their myriad hosts, generalist plant pathogens must tolerate a vast arsenal of plant specialized defense metabolites. To understand how host-specific metabolites influence plant-generalist pathogen interactions, we conducted a co-proteomic analysis of both Arabidopsis thaliana and Botrytis cinerea proteomes from the same samples during early infection. The Arabidopsis proteomic responses to Botrytis center around induction and suppression of defense metabolite pathways, particularly camalexin and glucosinolates. Several Botrytis proteins involved in key virulence pathways were induced within 32-48 hours, including potential defense metabolite detoxification proteins. Co-proteomic analysis using a panel of Arabidopsis genotypes with differing glucosinolate profiles revealed that disruptions to the glucosinolate pathway had broad changes on the Arabidopsis proteome, and that Botrytis induces specific proteins in response to presence/absence of Arabidopsis defense metabolites. Among the proteins that were induced quickly on infection and linked to the presence of glucosinolates, we validated a novel isothiocyanate hydrolase in Botrytis, BcSaxA, that catabolizes isothiocyanates in vitro. Gene expression data further indicated BcSaxA is expressed only in dicot hosts containing isothiocyanates. Our study describes a highly dynamic host proteome during infection with Botrytis and elucidates metabolite-specific infection strategies for a generalist pathogen.

plant biology↗

Host-Botrytis co-transcriptomics reveals finely tuned interactions with closely related legumes

Generalist pathogens infect diverse plant hosts, yet how these interactions differ across hosts is poorly understood. Here, we conduct a molecular analysis of a generalist pathogen interacting with closely related hosts. A co-transcriptomic framework is used to dissect host-pathogen interactions between the generalist necrotroph Botrytis cinerea and two closely related legume hosts, common bean (Phaseolus vulgaris) and cowpea (Vigna unguiculata). Using a diverse set of 72 Botrytis isolates, we quantified lesion development alongside host and pathogen gene expression. Although lesion formation was driven primarily by pathogen genetic variation, transcriptomic responses in both host and pathogen exhibited significant host x isolate interactions. This indicated that extensive, fine-scale transcriptional plasticity created similar disease outcomes. Botrytis genes showing host-specific expression were enriched for cell wall-modifying enzymes and some specialized metabolic genes, indicating greater host responsiveness of these core virulence mechanisms than previously appreciated. Co-expression network analysis in both host and pathogen further showed that in both organisms, gene membership for individual networks are restructured in response to genetic diversity. For example in Botrytis, we identify different sets of genes host-dependently co-expressing with a non-ribosomal peptide synthetase (NRPS) gene cluster, suggesting divergent functional deployment of the same virulence machinery across closely related hosts. Both legume species exhibited extensive isolate-dependent transcriptional reprogramming, with approximately two-thirds of expressed host genes responding to pathogen diversity. While conserved defense pathways such as jasmonate/ethylene signaling and phenylpropanoid metabolism were upregulated in both hosts, the specific genes in the networks differed markedly, highlighting lineage-specific rewiring of defense strategies. These results suggest that generalist pathogen success is underpinned by pervasive gene expression plasticity in both host and pathogen, allowing similar phenotypic outcomes to emerge from highly divergent molecular states. SummaryO_LIGeneralist pathogens infect diverse plant hosts, yet how these interactions differ across hosts is poorly understood. This study investigates how a generalist pathogen achieves successful infection across closely related hosts, and how these hosts respond. C_LIO_LIA co-transcriptomic approach was applied to interactions between 72 genetically diverse isolates of the fungal necrotroph Botrytis cinerea and two legume hosts, common bean and cowpea. Lesion development and host and pathogen gene expression were quantified. C_LIO_LILesion formation was primarily driven by pathogen genetic variation, yet both host and pathogen transcriptomes showed strong host x isolate interactions. Both host and pathogen balance conserved responses with finely tuned, host-specific mechanisms. Further, host-dependent transcriptional responses involve network modulation around a common core of genes in both host and pathogen. C_LIO_LIGeneralist pathogen success is underpinned by pervasive gene expression plasticity in both host and pathogen, allowing similar phenotypic outcomes to emerge from highly divergent molecular states. C_LI

plant biology↗

A multi-plant transcriptomic atlas reveals conserved and lineage specific defense architectures in response to Botrytis cinerea

Generalist pathogens pose a challenge to plant immunity by infecting diverse hosts while harboring extensive intraspecific genetic variation. Whether evolutionary distant plant lineages rely on a shared immune strategy or deploy distinct, lineage-specific defenses when confronted by these genetically variable members of the same pathogen species remains unresolved. Here, we employed a large-scale co-transcriptomic approach to map the immune landscape of ten diverse eudicot species infected with 72 genetically distinct Botrytis cinerea isolates. We identified a limited core of evolutionarily conserved defense orthologs, along with a vast landscape of lineage-specific transcriptional rewiring. While the broad physiological outcome such as metabolic reprogramming, cell wall modification, and suppression of growth-associated processes was shared across hosts, the regulatory pathways governing this were largely lineage-specific. Crucially, this immune landscape is dynamically shaped by pathogen diversity. Nearly three-quarters of host transcriptional responses were isolate-dependent, with the magnitude of defense activation defined by specific host-isolate combinations rather than a universal species-level response. Even host responses to shared virulence factors, including broadly expressed pathogens phytotoxins, were lineage specific. These findings show that plant immunity to generalist pathogens is built on conserved physiological outcomes executed through rapidly evolving, lineage-specific regulatory programs. This distinct regulatory architecture creates an immune landscape heavily modulated by specific host-isolate combinations, highlighting the necessity of integrating pathogen diversity into models of plant defense evolution and resistance breeding. Significance StatementAchieving durable, broad-spectrum crop protection remains difficult because plant immunity models often rely on limited species and overlook natural genetic diversity. Effective crop protection requires understanding how defense networks operate across diverse lineages. We tested this by measuring immune responses of ten phylogenetically diverse eudicots infected with 72 genetically distinct Botrytis cinerea isolates. We found that plants share conserved physiological defense outcomes achieved through highly divergent, lineage-specific regulatory networks. Host responses were strongly shaped by pathogen genetic diversity, with identical isolates eliciting different transcriptional responses in different hosts. This demonstrates that plant immunity quantitatively senses pathogen variation and disease outcomes emerge from specific host-isolate combinations. These findings explain the limits of resistance transfer and inform strategies for durable disease control.

genomics↗

Evaluating Regulatory Module Function within Mitochondrial Pyruvate Dehydrogenase Complex

Regulatory networks coordinate metabolism to control how plants adapt to biotic and abiotic stresses. This coordination can align transcriptional shifts across metabolic pathways using cis-regulatory elements shared across the enzyme genes within these pathways. While the role of transcription factors (TFs) in controlling this process across pathways is well known, less is known regarding the role of shared cis-regulatory elements across the genes in a pathway. Sharing cis-regulatory elements across the genes in an enzyme complex or pathway, can create coordinated regulation of the pathway by a TF. However, it is unclear if all the genes in a pathway or enzyme complex need to be fully coordinated for maximal function. For example, if one gene in an enzyme complex loses a cis-regulatory element, it may not alter the function of the enzyme complexes function if post-transcriptional or compensatory transcriptional changes are sufficient to balance the complex. To test how cis-modular membership shapes the function of an enzyme complex, we used CRISPR/Cas9 to abolish a common cis-regulatory element across the promoters of nine genes required for the mitochondrial pyruvate dehydrogenase complex (mtPDC). This complex is composed of three apoenzymes and is a central hub coordinating carbon flow between glycolysis and the tricarboxylic acid (TCA) cycle. Different combinations of these cis-element mutations were tested across the genes in the complex in Arabidopsis thaliana and the created genotypes were phenotyped for altered enzyme function using digital growth analysis, disease assays, metabolomics, and transcriptomics. This analysis revealed that mutating cis-element motifs of genes in this enzyme complex produced distinct phenotypes, displaying promoter-specific buffering and modularity.

molecular biology↗

Evaluating the Effectiveness of Data Reduction Techniques in QTL Mapping

Data reduction methods are frequently employed in large genomics and phenomics studies to extract core patterns, reduce dimensionality, and alleviate multiple testing effects. Principal component analysis (PCA), in particular, identifies the components that capture the most variance within omics datasets. While data reduction can simplify complex datasets, it remains unclear how the use of PCA impacts downstream analyses such as quantitative trait loci (QTL) or genome-wide association (GWA) approaches and their biological interpretation. In QTL studies, an alternative to data reduction is the use of post-hoc data summarization approaches, such as hotspot analysis, which involves mapping individual traits and consolidating results based on shared genomic locations. To evaluate how different analytical approaches may alter the biological insights derived from multi-dimensional QTL datasets, we compared individual trait hotspots with PCA-based QTL mapping using transcriptomic and metabolomic data from a structured recombinant inbred line population. Interestingly, these two approaches identified different genomic regions and genetic architectures. These findings suggest that mapping PCA-reduced data does not merely streamline analyses but may generate a fundamentally different view of the underlying genetic architecture compared to individual trait mapping and hotspot analysis. Thus, the use of PCA and other data reduction techniques prior to QTL or GWAS mapping should be carefully considered to ensure alignment with the specific biological question being addressed.

genetics↗

Combined generalist and host-specific transcriptional strategies enable host generalism in the fungal pathogen Botrytis cinerea

How generalist pathogens infect phylogenetically diverse hosts remains a central question in plant-pathogen biology. In particular, the extent to which broad host range is enabled by genetic variation versus transcriptional plasticity is unclear. To investigate how variation and plasticity contribute to generalism, we studied the generalist necrotrophic fungus Botrytis cinerea that infects more than 1,500 plant species. Using a cross-infection matrix of 72 B. cinerea isolates infected on 57 plant genotypes distributed across 15 eudicot species, we identified general and host-dependent fungal components of lesion formation. Transcriptome profiling at 48 hours post-inoculation revealed two distinct pathogen gene modules: (1) a set of general lesion-associated genes enriched in primary metabolism, showing similar expression across hosts but varied among isolates; and (2) a set of high-entropy, host specific-inducible genes, organized into distinct co-regulated modules that respond dynamically to specific host cues. Both gene sets were genomically dispersed, lacking structural clustering, and were under different levels of selective constraints. Our results demonstrate that B. cinerea employs a modular transcriptional strategy that integrates a core metabolic program along with a plastic, host-responsive regulatory network to achieve broad host colonization. This study presents the most comprehensive cross-species co-transcriptomic dataset to date for any fungal phytopathogen, highlighting transcriptional plasticity as a key mechanism underlying generalism in plant-fungal interactions. Moreover, the identification of conserved fungal gene targets across diverse hosts offers a foundation for developing broad-spectrum resistance strategies in multiple crops.

genomics↗

Shared selection and genetic architecture drive strikingly repeatable evolution in long-term experimental hybrid populations

The degree to which evolution repeats itself has implications regarding the major forces driving evolution and the potential for evolutionary biology to be a predictive (versus solely historical) science. To understand the factors that control evolutionary repeatability, we experimentally evolved four replicate hybrid populations of sunflowers at natural sites for up to 14 years and tracked ancestry across the genome. We found that there was very strong negative selection against introgressed ancestry in several chromosomes, but positive selection for introgressed ancestry in one chromosome. Further, the strength of selection was influenced by recombination rate. High recombination regions had lower selection against introgressed ancestry due to more frequent recombination away from incompatible backgrounds. Strikingly, evolution was highly parallel across replicates, with shared selection driving 88% of variance in introgressed allele frequency change. Parallel evolution was driven by both high levels of sustained linkage in introgressed alleles and strong selection on large-effect quantitative trait loci. This work highlights the repeatability of evolution through hybridization and confirms the central roles that natural selection, genomic architecture, and recombination play in the process.

evolutionary biology↗

The polygenic strategies of Botrytis cinerea for virulence across eight eudicot host species

Diverse qualitative and quantitative genetic architectures can successfully influence fungal virulence and host range. To model the quantitative genetic architecture of a generalist pathogen with an extensive host range, we conducted a genome-wide association study (GWAS) of the virulence of Botrytis cinerea across eight hosts. This approach revealed 4772 candidate genes, about one-third of the Botrytis genome, contributing to virulence with small effect sizes. The candidate genes are evenly spread across the core chromosomes with no indication of bipartite genomic architecture. The GWAS-identified polymorphisms and genes show that B. cinerea relies on genetic variants across hundreds of genes for infecting diverse hosts, with most genes influencing relatively few hosts. When pathogen genes associated to multiple hosts, they typically influenced more unrelated than related host species. Comparative genomics further suggested that the GWAS-identified genes are largely syntenic with other specialist Botrytis species and not novel to B. cinerea. Overall, B. cinereas generalist behavior is derived from the sum of the genome-wide genetic variation acting within gene networks that differentially coordinate the interaction with diverse hosts.

genetics↗

Leaf abaxial and adaxial surfaces differentially affect plant-fungal pathogen interactions

Eudicot plant species have bifacial leaves with each surface varying in a diversity of components, resulting in potentially different microhabitats for pathogens. We tested how Botrytis cinerea, a necrotroph fungal pathogen, interacts with the two different leaf surfaces across 16 crop species and 20 Arabidopsis genotypes. This showed that the abaxial surface is generally more susceptible to the pathogen than the adaxial surface. In Arabidopsis, the differential lesion area between leaf surfaces was associated to jasmonic acid (JA) and salicylic acid (SA) signaling and differential induction of defense chemistry. When infecting the adaxial surface, leaves mounted stronger defenses by producing more glucosinolates and camalexin defense compounds, partially explaining the differential susceptibility across surfaces. Testing a collection of 96 B. cinerea strains showed genetic heterogeneity of growth patterns, with a few strains preferring the adaxial surface while most are more virulent on the abaxial surface. Overall, we show that leaf-Botrytis interactions are complex with host-specific, surface-specific, and strain-specific behaviors. Within Arabidopsis, this mechanistically links to potential variation in JA/SA signaling across the two surfaces.

plant biology↗

Polygenic pathogen networks control of host and pathogen transcriptional plasticity in the Arabidopsis-Botrytis pathosystem

Bidirectional flow of information shapes the outcome of the host-pathogen interactions and depends on the genetics of each organism. Recent work has begun to use co-transcriptomic studies to shed light on this bidirectional flow, but it is unclear how plastic the co-transcriptome is in response to genetic variation in both the host and pathogen. To study co-transcriptome plasticity, we conducted transcriptomics using natural genetic variation in the pathogen, Botrytis cinerea, and large effect genetic variation abolishing defense signaling pathways within the host, Arabidopsis thaliana. We show that genetic variation in the pathogen has a greater influence on the co-transcriptome than mutations that abolish defense signaling pathways in the host. Genome wide association mapping using the pathogens genetic variation and both organisms transcriptomes allowed an assessment of how the pathogen modulates plasticity in response to the host. This showed that the differences in both organisms responses were linked to trans-eQTL hotspots within the pathogens genome. These hotspots control gene sets in either the host or pathogen and show differential allele sensitivity to the hosts genetic variation rather than qualitative host specificity. Interestingly, nearly all the trans-eQTL hotspots were unique to the host or pathogen transcriptomes. In this system of differential plasticity, the pathogen mediates the shift in the co-transcriptome more than the host.

plant biology↗

Epistatic Transcription Factor Networks Differentially Modulate Arabidopsis Growth and Defense

Plants integrate internal and external signals to finely coordinate growth and defense allowing for maximal fitness within a complex environment. One common model for the relationship between growth and defense is a trade-off model in which there is a simple negative interaction between growth and defense theoretically driven by energy costs. However, there is a developing consensus that the coordination of growth and defense likely involves a more conditional and intricate connection. To explore how a transcription factor network may coordinate growth and defense, we used high-throughput phenotyping to measure growth and flowering in a set of single and pairwise mutants previously linked to the aliphatic glucosinolate defense pathway. Showing the link between growth and aliphatic glucosinolate defense, 17 of the 20 tested TFs significantly influence plant growth and/or flowering time. These effects were conditional upon the environment, age of the plant and more critically varied amongst the phenotypes when using the same genotype. The phenotypic effects of the TF mutants on SC GLS accumulation and on growth did not display a simple correlation, supporting the coordination model. We propose that large transcription factor networks create a system to integrate internal and external signals and separately modulate growth and the accumulation of the defensive aliphatic GLS.

plant biology↗

A generalist pathogen view of plant evolution

Botrytis cinerea is a polyphagous fungal pathogen that causes necrotic disease on more than a thousand known hosts widely spread across the plant kingdom. While it is known that quantitative resistance in the host and quantitative virulence in the pathogen largely mediate this pathosystem, how this pathogen interacts with the extensive host diversity is unknown. Does this pathogen have quantitative virulence efficiency on all hosts or individual solutions for each host? To address this question, we generated an infectivity matrix of 98 strains of Botrytis cinerea on 90 genotypes representing eight host plants. This experimental infectivity matrix showed that the predominant sources of quantitative variation are between host species and among pathogen strains. Furthermore, the eight eudicot hosts interacted individually with Botrytis cinerea strains independently of the evolutionary relatedness between hosts. An additive quantitative model can explain the complexity of these interactions in which Botrytis host specificity and general virulence have distinct polygenic architectures.

plant biology↗

Admixture mapping in interspecific Populus hybrids identifies classes of genomic architectures for phytochemical, morphological and growth traits

The genomic architecture of functionally important traits is key to understanding the maintenance of reproductive barriers and trait differences when divergent populations or species hybridize. We conducted a Genome-Wide Association Study (GWAS) to study trait architecture in natural hybrids of two ecologically divergent Populus species. We genotyped 472 seedlings from a natural hybrid zone of Populus alba and P. tremula for genome-wide markers from reduced representation sequencing, phenotyped the plants in common gardens for 46 phytochemical (phenylpropanoid), morphological, and growth traits, and used a Bayesian polygenic model for mapping. We detected three classes of genomic architectures: (1) traits with finite, detectable associations of genetic loci with phenotypic variation in addition to highly polygenic heritability, (2) traits with indications for polygenic heritability only, (3) traits with no detectable heritability. For class (1), we identified genome regions with plausible candidate genes for phenylpropanoid biosynthesis or its regulation, including MYB transcription factors and glycosyl transferases. GWAS in natural, recombinant hybrids represents a promising step towards resolving the genomic architecture of phenotypic traits in long-lived species. This facilitates the fine-mapping and subsequent functional characterization of genes and networks causing differences in hybrid performance and fitness.

evolutionary biology↗

Combining Digital Imaging and Genome Wide Association Mapping to Dissect Uncharacterized Traits in Plant/Pathogen Interactions

Plant resistance to generalist pathogens with broad host ranges, such as Botrytis cinerea, is typically quantitative and highly polygenic. Recent studies have begun to elucidate the molecular genetic basis underpinning plant-pathogen interactions using commonly measured traits including lesion size and/or pathogen biomass. Yet with the advent of digital imaging and phenomics, there are a large number of additional resistance traits available to study quantitative resistance. In this study, we used high-throughput digital imaging analysis to investigate previously uncharacterized visual traits of plant-pathogen interactions related disease resistance using the Arabidopsis thaliana/Botrytis cinerea pathosystem. Using a large collection of 75 visual traits collected from every lesion, we focused on lesion color, lesion shape, and lesion size, to test how these aspects of the interaction are genetically related. Using genome wide association (GWA) mapping in A. thaliana, we show that lesion color and shape are genetically separable traits associated with plant-disease resistance. Using defined mutants in 23 candidate genes from the GWA mapping, we could identify and show that novel loci associated with each different plant-pathogen interaction trait, which expands our understanding of the functional mechanisms driving plant disease resistance.\n\nSummaryDigital imaging allows the identification of genes controlling novel lesion traits.

plant biology↗