Biology subjects

Cicconardi, F.

Publications and source records attributed to Cicconardi, F..

14 recordsLinked to original sources

bioRxiv2026

Characterisation of cis-regulatory elements and genome wide enhancer prediction in Heliconiini butterflies

The large-scale annotation and spatiotemporal characterisation of cis-regulatory promoters and enhancers remain limited in many non-model organisms, largely due to fragmented availability of multiomic datasets across species and developmental conditions. Heliconius butterflies, which have been used to investigate wing development and patterning, have provided key insights into the genetic architecture underlying morphological diversification and adaptive pigmentation. Despite this, we are limited in what we understand of how cis-regulatory dynamics drive phenotypic wing patterning variation. Here, by re-analysing and re-evaluating available data, we present a comprehensive new description of a cis-regulatory atlas for Heliconius wing development by integrating chromatin accessibility dynamics (ATAC-seq) with histone modification profiling (H3K27ac, H3K4me3). High-confidence peak sets enabled robust annotation of promoters and enhancers, revealing regulatory distributions consistent with canonical regulatory organisation. Promoters displayed comparatively higher conservation of transcription factor motifs, whilst enhancers exhibited greater sequence diversity, broader genomic distributions and more variable motif repertoires. Comparative analysis revealed cis-regulatory divergence to be primarily temporal, with substantial chromatin remodelling and differential histone enrichment across development, while regulatory dynamics were highly conserved between forewing and hindwing tissues. Extending these experimentally characterised regulatory landscapes, a convolutional neural network was trained for sequence-based enhancer prediction, achieving test AUROC and AUPRC values of 0.909 and 0.913, respectively. Predictions across 34 Heliconiini assemblies displayed significant enrichment within independently characterised accessible chromatin and conserved non-exonic elements (CNEEs), supporting their putative regulatory function. Overall, this study provides a large-scale characterisation of cis-regulatory dynamics and extends enhancer annotation across Heliconiini, providing a foundation for comparative investigations of regulatory conservation and diversification.

genomics↗

bioRxiv2026

Ancient Rapid Radiation Underlies Persistent Phylogenomic Conflict in Early Collembola Diversification

Collembola (springtails) are among the most abundant and ecologically important soil arthropods, representing one of the oldest extant terrestrial hexapod lineages, with a fossil record extending to the early Devonian. Despite their relevance, phylogenetic relationships among the four extant orders (Entomobryomorpha, Poduromorpha, Symphypleona, and Neelipleona) have remained unresolved for over two decades. Here, we present the most comprehensive phylogenomic analysis of Collembola to date, comprising 1,127 single-copy orthologues from 145 taxa representing 19 families. To improve orthology inference, we developed a novel HMM-based filtering pipeline that significantly reduced hidden paralogy in BUSCO-derived datasets. Across multiple dataset configurations, gene-jackknife replicates, and various maximum-likelihood analyses, we consistently recovered Poduromorpha as the earliest-diverging lineage. Coalescent-based methods instead highlighted discordant arrangements characterised by extremely short internal branches and low quartet support, a pattern consistent with pervasive incomplete lineage sorting and reticulate evolutionary history. We further dissected the phylogenetic signal by exhaustively evaluating all possible inter-order topological arrangements, both on the full concatenated dataset and gene-by-gene, to identify the most phylogenetically informative loci. These analyses rejected the great majority of previously proposed hypotheses, narrowing support to only two statistically indistinguishable topologies (T11 and T4), with the Poduromorpha-first arrangement consistently favoured across both site-homogeneous and site-heterogeneous substitution models. Finally, with molecular dating, we estimated the origin of crown Collembola in the Early Devonian, with the diversification of the extant orders in the Carboniferous. Several extant genera were estimated to be older than many currently recognized families, highlighting the exceptional evolutionary persistence of springtail lineages and suggesting that lineage longevity should be considered when interpreting higher-level taxonomic diversity.

evolutionary biology↗

bioRxiv2026

Genomic insights into bacterial isolates dominating honeypot ant crop microbiomes reveal metabolically distinct Fructilactobacillus sp.

Honeypot ants engage in a convergently-evolved phenotype called repletism, where specialized workers expand their crops and gasters to store vast amounts of food internally. They then store that food for months to support colonies during times of food scarcity. This fascinating phenotype is not well-understood and very little is known about the microbial interactions happening within the fructose-rich replete crop. Previous research using amplicon sequencing showed that Fructilactobacillus makes up nearly 100% of the crop microbiomes of Myrmecocystus mexicanus repletes. This striking result and successful isolation of those strains led to the present investigation into the phylogenetic diversity of these strains and any clues to the nature of the symbiotic relationship between them and the ant host. We find that the isolates from these repletes represented two evolutionary lineages, both most closely related to F. fructivorans. One of those lineages was also found to be phylogenetically and metabolically distinct from all other Fructilactobacillus reference genomes used in this study. This discovery in a genus of bacteria that are highly relevant for fermented human foods and will also lay the groundwork for future understanding of the convergent evolutionary mechanisms of repletism in ants. 3. Impact statementThese analyses add to the literature by identifying a new microbe within a genus that is relevant to food systems. In addition, the host phenotype is convergently evolved and likely microbe-mediated (or at least highly exposed). Understanding this system allows for the testing of ideas of coevolutionary hypotheses with natural replicates. We expect interest to come from food safety and probiotics researchers, evolutionary biologists that think about the impacts of microbes, microbial ecologists interested in novel systems, and those interested in bacteria that may display unique metabolic possibilities. This output allows for the clear future examination of this system with many clear hypotheses. Our analysis allows for the creation of a new and unique model system of host-microbe symbiosis. 4. Data summaryNew genomes assembled in this work can be found under BioProject ID PRJNA1449409. https://www.ncbi.nlm.nih.gov/bioproject/?term=PRJNA1449409. Reference bacterial genomes were obtained from NCBI at the following accession numbers: SAMN20557570, SAMN28081009, SAMN28081010, SAMN02597458, SAMN43111088, SAMN28081011, SAMN28535231, SAMN04505734, SAMN28081013, SAMN33452149, SAMN37926504, SAMN02849426, SAMN02470196, SAMN02369432, SAMN02797779, SAMN02797782, SAMN02797768, SAMN09762388, SAMN12785275, SAMEA117660288. The honeypot ant genome was obtained from SAMN37666067. Raw proteomics files will be uploaded to ProteomeXchange with a unique identifier upon manuscript acceptance.

genomics↗

bioRxiv2025

Polygenic basis of incipient reproductive isolation in hybridizing populations is revealed by pangenomic and epigenetic divergence

Incipient reproductive isolation in the presence of gene flow has traditionally been attributed to a small number of major-effect loci under strong selection, representing only a minor fraction of the genome. Using the Heliconius erato adaptive radiation--a butterfly species with populations at varying levels of genetic differentiation--we employed a pan-genome approach to investigate genome-wide mutational divergence, epigenetic changes, and structural variation contributing to divergence. In contrast to earlier studies that identified only a few highly divergent loci, our high-resolution analysis reveals widespread divergence across the genome, suggesting polygenic barriers to gene flow. Epigenetic divergence assessed using ATAC-seq, highlights population-specific differences in chromatin accessibility, which may reflect SNPs distribution or precede genetic differentiation by reshaping regulatory landscapes. We also identified new structural variants, including polymorphic indels in open chromatin, that further associate with genomic divergence. Together, these findings show that the genomic landscape of H. erato is shaped by a dynamic interplay of mutational changes, epigenetic modifications, and structural variation. We emphasize the role of developmental, behavioral, and ecological adaptations and provide a broader perspective on the functional genetic basis of genome-wide divergence in Heliconius. This emerging polygenic framework offers a more detailed understanding of how reproductive barriers evolve under ongoing gene flow.

evolutionary biology↗

bioRxiv2025

Gene loss associated with plasticity-first evolution in Heliconius butterflies

Phenotypic plasticity occurs when a genotype can produce more than one phenotype under different environmental conditions. Genetic accommodation allows plastic phenotypes to be tuned to new environments and eventually to even be lost. The colourful and toxic Heliconiini butterflies have biochemical plasticity - they either sequester their cyanogenic glucosides (CG) from their larval hostplant or biosynthesize them when compounds for sequestration are not available. Here, we trace the evolution of CG biosynthesis in Heliconiini butterflies, a fundamental component of this biochemical plasticity. We first reconstructed the evolutionary history of biochemical plasticity in Heliconiini butterflies using chemical data from over 700 individuals, demonstrating that plasticity was ancestral in the tribe and further lost in a few clades, such as the Sapho clade that only sequester CGs. In lepidopterans, CG biosynthesis has been fully characterized in the moth Zygaena filipendulae, and is composed of CYP405A2, CYP332A3, and UGT33A1. Thus, we CRISPR-edited the gene CYP405 in Heliconius erato, and confirmed that CYP405-knockout caterpillars do not biosynthesize CGs. We identified the CYP405s and CYP332s in other lepidopterans and found that both genes were independently co-opted into CG biosynthesis in the Heliconiinae butterflies and Zygaena moths. Most lepidopterans have a CYP332, but CYP405 is mostly restricted to butterflies and has been duplicated in all Heliconius species. Although several CYP405 copies were found in the Sapho clade, they lack structurally important P450 domains, which explain they loss of biochemical plasticity via specialization in CG sequestration. Our findings represent one of the few examples of plasticity-first evolution in which the genetic mechanisms associated with its accommodation/assimilation are known.

evolutionary biology↗

bioRxiv2025

Evolution of the olfactory system during the radiation of Heliconiini butterflies

Sensory system evolution plays a crucial role in shaping species interactions with their environment, yet the extent to which olfactory system diversity reflects ecological and evolutionary pressures at a macroevolutionary scale remains unclear. Here, we investigate the evolution of the olfactory system across the Heliconiini butterfly tribe, an ecologically diverse but closely related group. Using a comparative approach, we examined variation in antennal lobe morphology and its constituent structures, the glomeruli and antennal lobe hub, as well as olfactory receptor repertoires across species. We found that antennal lobe size variation is driven by independent shifts in glomerular and antennal lobe hub volumes, with species-specific differences occurring against a backdrop of broader phylogenetic stability. While no direct associations with ecological traits were observed, certain species showed large expansions in total glomerular volume and olfactory receptor numbers, warranting further investigation into unmeasured ecological or behavioural factors. Additionally, comparisons between wild-caught and insectary-reared individuals revealed a surprising pattern of developmental plasticity, with antennal lobe hub volumes increasing and glomeruli volumes decreasing in captivity, highlighting the influence of environmental conditions on neural development. These findings suggest that olfactory evolution in Heliconiini is shaped by both evolutionary divergence and developmental plasticity, emphasizing the need to integrate phylogenetic, ecological, and developmental perspectives to fully understand sensory system adaptation.

evolutionary biology↗

bioRxiv2025

Convergent molecular evolution associated with repeated transition to gregarious larval behaviour in Heliconiini

Collective behaviour forms the basis for many anti-predator strategies. Within Lepidoptera, larval gregariousness has evolved convergently across many phylogenetically disparate lineages. While the selection pressures shaping variation in larval social behaviours are well investigated, much less is known about the mechanisms that control social attraction and behavioural coordination. Similarly, little is known about how secondary selection pressures associated with social living shape genome evolution. Here, using genomic data for over 60 species from an adaptive radiation of Neotropical butterflies, the Heliconiini, in which gregarious behaviour has evolved repeatedly, we explore the molecular basis of repeated convergent shifts towards gregarious larvae. We focus on three main areas of genomic evolution: differential selection on homologous genes, accelerated rates of evolution on non-coding regions of key genes, and differential gene expression in the brains of solitary and gregarious larvae. We identify strong signatures of convergent molecular evolution, on both coding and non-coding loci, in Heliconiini lineages which evolved gregarious behaviour. Molecular convergence is also detected at the transcriptomic level in larval brains, suggesting convergent shifts in gene regulation in neural tissue. Among loci showing strong signals of convergent evolution in gregarious lineages, we identify several strong candidates linked to neural activity, feeding behaviour, and immune pathways. Our results suggest sociality profoundly changes the selection pressures acting on multiple physiological, immunological and behavioural traits.

evolutionary biology↗

bioRxiv2023

Multiple axes of visual system diversity in Ithomiini, an ecologically diverse tribe of mimetic butterflies

The striking structural variation seen in arthropod visual systems can be explained by the overall quantity and spatio-temporal structure of light within habitats coupled with developmental and physiological constraints. However, little is currently known about how fine-scale variation in visual structures arise across shorter evolutionary and ecological scales. In this study, we characterise patterns of interspecific (between species), intraspecific (between sexes) and intraindividual (between eye regions) variation in the visual system of four ithomiine butterfly species. These species are part of a diverse 26-Myr-old Neotropical radiation where changes in mimetic colouration are associated with fine-scale shifts in ecology, such as microhabitat preference. By using a combination of selection analyses on visual opsin sequences, in-vivo ophthalmoscopy, micro-computed tomography (micro-CT), immunohistochemistry, confocal microscopy, and neural tracing, we quantify and describe physiological, anatomical, and molecular traits involved in visual processing. Using these data, we provide evidence of substantial variation within the visual systems of Ithomiini, including: i) relaxed selection on visual opsins, perhaps mediated by habitat preference, ii) interspecific shifts in visual system physiology and anatomy, and iii) extensive sexual dimorphism, including the complete absence of a butterfly-specific optic neuropil in the males of some species. We conclude that considerable visual system variation can exist within diverse insect radiations, hinting at the evolutionary lability of these systems to rapidly develop specialisations to distinct visual ecologies, with selection acting at both the perceptual, processing, and molecular level. Summary statementPhysiological, anatomical, and molecular evidence of extensive visual system variation within a diverse butterfly radiation, hinting at the lability of visual systems to evolve specialisations to distinct visual environments.

evolutionary biology↗

bioRxiv2022

Rapid expansion and visual specialization of learning and memory centers in Heliconiini butterflies

How do neural systems evolve to support new behaviors? Changes in the abundance and diversity of neural cell types, and their connectivity, shape brain composition and provide the substrate for behavioral variation. We describe a striking example of neural elaboration in an ecologically diverse tribe of Heliconiini butterflies. By building extensive new datasets of neural traits across the tribe, we identify major bursts in the size and cellular composition of the mushroom bodies, central brain structures essential for learning and memory. These expansion events are associated with increased innervation form visual centers and coincide with enhanced performance in multiple cognitive assays. This suite of neural and cognitive changes is likely tied to the emergence of derived foraging behaviors, facilitated by localized specialization of neural networks. One-Sentence SummaryMajor shifts in brain composition and behavior in butterflies with unique foraging and dietary behaviors.

evolutionary biology↗

bioRxiv2022

Evolutionary dynamics of genome size and content during the adaptive radiation of Heliconiini butterflies

Heliconius butterflies, a speciose genus of Mullerian mimics, represent a classic example of an adaptive radiation that includes a range of derived dietary, life history, physiological and neural traits. However, key lineages within the genus, and across the broader Heliconiini tribe, lack genomic resources, limiting our understanding of how adaptive and neutral processes shaped genome evolution during their radiation. We have generated highly contiguous genome assemblies for nine new Heliconiini, 29 additional reference-assembled genomes, and improve 10 existing assemblies. Altogether, we provide a major new dataset of annotated genomes for a total of 63 species, including 58 species within the Heliconiini tribe. We use this extensive dataset to generate a robust and dated heliconiine phylogeny, describe major patterns of introgression, explore the evolution of genome architecture, and the genomic basis of key innovations in this enigmatic group, including an assessment of the evolution of putative regulatory regions at the Heliconius stem. Our work illustrates how the increased resolution provided by such dense genomic sampling improves our power to generate and test gene-phenotype hypotheses, and precisely characterize how genomes evolve.

evolutionary biology↗

bioRxiv2022

A butterfly pan-genome reveals a large amount of structural variation underlies the evolution of chromatin accessibility

Despite insertions and deletions being the most common structural variants (SVs) found across genomes, not much is known about how much these SVs vary within populations and between closely related species, nor their significance in evolution. To address these questions, we characterized the evolution of indel SVs using genome assemblies of three closely related Heliconius butterfly species. Over the relatively short evolutionary timescales investigated, up to 18.0% of the genome was composed of indels between two haplotypes of an individual H. charithonia butterfly and up to 62.7% included lineage-specific SVs between the genomes of the most distant species (11 Mya). Lineage-specific sequences were mostly characterized as transposable elements (TEs) inserted at random throughout the genome and their overall distribution was similarly affected by linked selection as single nucleotide substitutions. Using chromatin accessibility profiles (i.e., ATAC-seq) of head tissue in caterpillars to identify sequences with potential cis-regulatory function, we found that out of the 31,066 identified differences in chromatin accessibility between species, 30.4% were within lineage-specific SVs and 9.4% were characterized as TE insertions. These TE insertions were localized closer to gene transcription start sites than expected at random and were enriched for several transcription factor binding site candidates with known function in neuron development in Drosophila. We also identified 24 TE insertions with head-specific chromatin accessibility. Our results show high rates of structural genome evolution that were previously overlooked in comparative genomic studies and suggest a high potential for structural variation to serve as raw material for adaptive evolution.

genomics↗

bioRxiv2021

The effects of chromosome fusions on genetic diversity and evolutionary turnover of functional loci consistently depends on chromosome size

Major changes in chromosome number and structure are linked to a series of evolutionary phenomena, including intrinsic barriers to gene flow or suppression of recombination due to chromosomal rearrangements. However, chromosome rearrangements can also affect the fundamental dynamics of molecular evolution within populations by changing relationships between linked loci and altering rates of recombination. Here, we build chromosome-level assembly Eueides isabella and, together with the chromosome-level assembly of Dryas iulia, examine the evolutionary consequences of multiple chromosome fusions in Heliconius butterflies. These assemblies pinpoint fusion points on 10 of the 21 autosomal chromosomes and reveal striking differences in the characteristics of fused and unfused chromosomes. The ten smallest autosomes in D. iulia and E. isabella, which have each fused to a longer chromosome in Heliconius, have higher repeat and GC content, and longer introns than predicted by their chromosome length. Following fusion, these characteristics change to become more in line with chromosome length. The fusions also led to reduced diversity, which likely reflects increased background selection and selection against introgression between diverging populations, following a reduction in per-base recombination rate. We further show that chromosome size and fusion impact turnover rates of functional loci at a macroevolutionary scale. Together these results provide further evidence that chromosome fusion in Heliconius likely had dramatic effects on population level processes shaping rates of neutral and adaptive divergence. These effects may have impacted patterns of diversification in Heliconius, a classic example of an adaptive radiation.

evolutionary biology↗

bioRxiv2020

Unveiling diffusion pattern and structural impact of the most invasive SARS-CoV-2 spike mutation

Starting in Wuhan, China, SARS-CoV-2 epidemics quickly propagated worldwide in less than three months, geographically sorting genomic variants in newly established propagules of infections. Stochasticity in transmission within and between countries and/or actual advantage in virus transmissibility could explain the high frequency reached by some genomic variants during the course of the outbreak. Using a suite of statistical, population genetics, and theoretical approaches, we show that the globally most represented spike protein variant (i.e., the G clade, A [->] G nucleotide change at genomic position 23,403; D [->] G amino acid change at spike protein position 614) i) underwent a significant demographic expansion in most countries not explained by stochastic effects or enhanced pathogenicity; ii) affects the spike S1/S2 furin-like site increasing its conformational plasticity (short range effect), and iii) modifies the internal motion of the receptor-binding domain affecting its cross-connection with other functional domains (long-range effect). Our study unambiguously links the spread of the G614 with a non-random process, and we hypothesize that this process is related to the selective advantage produced by a specific structural modification of the spike protein. We conclude that the different conformation of the S1/S2 proteolytic site is at the basis of the higher transmission rate of this invasive SARS-CoV-2 variant, and provide structural information to guide the design of selective and efficient drugs.

evolutionary biology↗

bioRxiv2019

Genomic signature of shifts in selection in a sub-alpine ant and its physiological adaptations

Understanding how organisms adapt to extreme environments is fundamental and can provide insightful case studies for both evolutionary biology and climate-change biology. Here, we take advantage of the vast diversity of lifestyles in ants to identify genomic signatures of adaptation to extreme habitats such as high altitude. We hypothesised two parallel patterns would occur in a genome adapting to an extreme habitat: i) strong positive selection on genes related to adaptation and, ii) a relaxation of previous purifying selection. We tested this hypothesis by sequencing the high-elevation specialist Tetramorium alpestre and four other phylogenetic related species. In support of our hypothesis, we recorded a strong shift of selective forces in T. alpestre, in particular a stronger magnitude of diversifying and relaxed selection when compared to all other ants. We further disentangled candidate molecular adaptations in both gene expression and protein-coding sequence that were identified by our genome-wide analyses. In particular, we demonstrate that T. alpestre has i) a derived level of expression for stv and other heat-shock proteins in chill shock tests, and ii) enzymatic enhancement of Hex-T1, a rate-limiting regulatory enzyme that controls the entry of glucose into the glycolytic pathway. Together, our analyses highlight the adaptive molecular changes that support colonisation of high-altitude environments.

evolutionary biology↗