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Widespread decoupling between abundance and genetic diversity, and strong local genetic structuring in marine unicellular eukaryotes

How abundance, connectivity, and genetic diversity covary across space remains a central question in evolutionary biology. Using a global environmental DNA metabarcoding dataset targeting mitochondrial cytochrome c oxidase subunit I (COI), we examined relationships among metabarcoding-derived abundance, nucleotide diversity, and spatial genetic connectivity across marine eukaryotes. We found a widespread but incomplete decoupling between abundance and genetic diversity, particularly in unicellular lineages. By contrast, distance-decay patterns in abundance, diversity, and connectivity were broadly similar across eukaryotes and more strongly associated with cellularity. Focusing on the Iberian Peninsula, we reconstructed locality-centred haplotype networks to quantify the spatial organization of intraspecific genetic variation. Unicellular lineages showed stronger local genetic structuring, whereas multicellular organisms exhibited clearer large-scale biogeographic partitioning. Overall, our results reveal shared biogeographic organization across eukaryotes while demonstrating that abundance-based biodiversity metrics alone cannot fully capture evolutionary and demographic processes, especially in microbial organisms.

ecology↗

Three annotated tiger beetle genomes (Coleoptera, Adephaga, Cicindelidae)

Advances and accessibility to next-generation technology provide opportunities to sequence non-model organisms. Despite this increase in whole-genome sequencing data, annotation and the production of reference genomes remain limited. Reference genomes are a critical tool for a variety of studies in evolutionary biology, functional genomics, and conservation genetics. Tiger beetles (Cicindelidae) are a diverse and globally distributed family of beetles that serve as bioindicator taxa and flagship species for insect conservation. Here, we report highly complete, contiguous, and annotated genome assemblies representing draft reference genomes for three species of tiger beetle spanning the phylogeny. These draft reference genomes are for Audouins night-stalking tiger beetle, Omus audouini; the montane giant tiger beetle, Amblycheila baroni; and the western red-bellied tiger beetle, Cicindelidia sedecimpunctata. Article SummaryTiger beetles are a charismatic group with [~]3000 species distributed globally. Despite their popularity among insect enthusiasts and their role as bioindicators of ecosystem health, the group currently lacks a reference genome. This article outlines genome assembly and annotation for three tiger beetle species that span evolutionary relationships within the lineage. Quality control analyses show that the assemblies are reference quality and demonstrate high contiguity, completeness, and accuracy. The resulting draft genome annotations will be a valuable resource for scientific endeavors and allow for continued research on tiger beetles and their allies.

genomics↗

A Curated Genome-Scale Nucleotide Diversity Panel of Non-Human Primates

BackgroundPrimates constitute one of the most phylogenetically and ecologically diverse Eutherian mammalian orders, with a central role in advancing our knowledge of human evolution, speciation processes and conservation biology. While thousands of whole-genome sequences have been generated across a multitude of primate taxa, discrepancies in data processing - particularly the lack of ploidy-aware variant calling in sex-linked regions - have limited the utility of existing datasets for large-scale comparative analyses. ResultsHere, we utilized publicly available short-read sequencing data of non-human primates, recently published primate genome assemblies and a ploidy-aware variant calling procedure to generate a genome-scale nucleotide diversity panel comprising 3,240 individuals from 269 species and 71 genera. To further facilitate cross-species comparisons, we generated a multiple-genome alignment of primate assemblies used for variant calling. ConclusionThis curated resource of non-human primate diversity provides a foundation for future research in primate evolutionary biology, speciation, and sex chromosome evolution (https://pure.au.dk/portal/en/datasets/primate-diversity-panel/).

genomics↗

Chi.Bio: An open-source automated experimental platform for biological science research

The precise characterisation and manipulation of in vivo biological systems is critical to their study.1 However, in many experimental frameworks this is made challenging by non-static environments during cell growth,2, 3 as well as variability introduced by manual sampling and measurement protocols.4 To address these challenges we present Chi.Bio, a parallelised open-source platform that offers a new experimental paradigm in which all measurement and control actions can be applied to a bulk culture in situ. In addition to continuous-culturing capabilities (turbidostat functionality, heating, stirring) it incorporates tunable light outputs of varying wavelengths and spectrometry. We demonstrate its application to studies of cell growth and biofilm formation, automated in silico control of optogenetic systems, and readout of multiple orthogonal fluorescent proteins. By combining capabilities from many laboratory tools into a single low-cost platform, Chi.Bio facilitates novel studies in synthetic, systems, and evolutionary biology, and broadens access to cutting-edge research capabilities.

bioengineering↗

Pleistocene climate change and the formation of regional species pools

This preprint has been reviewed and recommended by Peer Community In Evolutionary Biology (https://doi.org/10.24072/pci.evolbiol.100053). Despite the description of bioregions dates back from the origin of biogeography, the processes originating their associated species pools have been seldom studied. Ancient historical events are thought to play a fundamental role in configuring bioregions, but the effects of more recent events on these regional biotas are largely unknown. We use a network approach to identify regional and sub-regional faunas of European Carabus beetles, and analyse the effects of dispersal barriers, niche similarities and phylogenetic history on their configuration. We identify a transition zone matching the limit of the ice sheets at Last Glacial Maximum. While southern species pools are mostly separated by dispersal barriers, in the north species are mainly sorted by their environmental niches. Strikingly, most phylogenetic structuration of Carabus faunas occurred during the Pleistocene. Our results show how extreme recent historical events -such as Pleistocene climate cooling, rather than just deep-time evolutionary processes, can profoundly modify the composition and structure of geographic species pools.

ecology↗

The genetic architecture of local adaptation is historically contingent

Revealing the genetic basis of local adaptation is a common goal of evolutionary biology, but despite theoretical progress, general expectations for the genetic architecture of local adaptation are still unclear. Theoretical analyses usually model simplified ecologies or simplified genetic architectures of adaptive traits, so the interplay of these factors is missing from our understanding. In this study, we use simulations to explore how the interplay of ecological and genetic parameters influences the evolution and genetic architecture of local adaptation. With these simulations, we ask: i) What are the features of alleles that made the largest contribution to local adaptation, and how are they affected by polygenicity of adaptive traits, migration rates, demographic history, and the spatial pattern of the environment? And ii) does allele age moderate the confounding effect from population structure in genotype-environmental associations (GEA)? We find that the frequency, number, and phenotypic effect size of locally adaptive alleles are sensitive to trait polygenicity and demographic history, and that these factors shape the evolutionary dynamics of local adaptation. We find that population expansions can leave legacies in the genetic architecture of local adaptation, reducing the expected number of adaptive alleles relative to models with constant population size, and this effect is long-lasting. Compared to range expansion, other ecological variables known to affect the genetic basis of local adaptation had limited effects. Finally, allele age moderated the confounding effect of population structure and modified the causal effect of environmental variables on genotypes. Alleles that arose around the time of environmental changes often made large contributions to local adaptation, but young alleles often had the highest false positive rates and were the most common age category. We describe how incorporating allele age and its interactions with population structure and environmental variables may increase the sensitivity and specificity of GEA analysis. Overall, this work demonstrates the critical importance that a species demographic history can have on its genetic architecture of local adaptation.

genetics↗

Genetic transformation of the frog-killing chytrid fungus Batrachochytrium dendrobatidis

Batrachochytrium dendrobatidis (Bd), a causative agent of chytridiomycosis, is decimating amphibian populations around the world. Bd belongs to the chytrid lineage, a group of early-diverging fungi that are widely used to study fungal evolution. Like all chytrids, Bd develops from a motile form into a sessile, growth form, a transition that involves drastic changes in its cytoskeletal architecture. Efforts to study Bd cell biology, development, and pathogenicity have been limited by the lack of genetic tools with which to test hypotheses about underlying molecular mechanisms. Here, we report the development of a transient genetic transformation system for Bd. We used electroporation to deliver exogenous DNA into Bd cells and detected transgene expression for up to three generations under both heterologous and native promoters. We also adapted the transformation protocol for selection using an antibiotic resistance marker. Finally, we used this system to express fluorescent protein fusions and, as a proof of concept, expressed a genetically encoded probe for the actin cytoskeleton. Using live-cell imaging, we visualized the distribution and dynamics of polymerized actin at each stage of the Bd life cycle, as well as during key developmental transitions. This transformation system allows, for the first time, direct testing of key hypotheses regarding mechanisms of Bd pathogenesis. This technology also paves the way for answering fundamental questions of chytrid cell, developmental, and evolutionary biology.

cell biology↗

Algorithms to reconstruct past indels: the deletion-only parsimony problem

Ancestral sequence reconstruction is an important task in bioinformatics, with applications ranging from protein engineering to the study of genome evolution. When sequences can only undergo substitutions, optimal reconstructions can be efficiently computed using well-known algorithms. However, accounting for indels in ancestral reconstructions is much harder. First, for biologically-relevant problem formulations, no polynomial-time exact algorithms are available. Second, multiple reconstructions are often equally parsimonious or likely, making it crucial to correctly display uncertainty in the results. Here, we consider a parsimony approach where any indel event has the same cost, irrespective of its size or the branch where it occurs. We thoroughly examine the case where only deletions are allowed, while addressing the aforementioned limitations. First, we describe an exact algorithm to obtain all the optimal solutions. The algorithm runs in polynomial time if only one solution is sought. Second, we show that all possible optimal reconstructions for a fixed node can be represented using a graph computable in polynomial time. While previous studies have proposed graph-based representations of ancestral reconstructions, this result is the first to offer a solid mathematical justification for this approach. Finally we discuss the relevance of the deletion-only case for the general case. Author summaryAn exciting frontier in evolutionary biology is the ability to reconstruct DNA or protein sequences from species that lived in the distant past. By analyzing sequences from present-day species, we aim to infer the sequences of their common ancestors --a process known as ancestral sequence reconstruction. This task has far-reaching applications, such as resurrecting ancient proteins and studying the biology of extinct organisms. However, a significant challenge remains: the lack of well-established methods for inferring past deletions and insertions ---mutations that remove or add segments of genetic code. In this paper, we present algorithms that lay the groundwork for addressing this gap. We show that finding the reconstructions involving only deletion events, while minimizing their number, can be done efficiently. Additionally, we show that all optimal solutions can be represented using specialized graphs. While previous studies have proposed graph-based representations of ancestral reconstructions, we are the first to provide a rigorous mathematical foundation for the use of these graphs.

bioinformatics↗

Novel compartmental models of infectious disease transmission

For decades, mathematical models of disease transmission have provided researchers and public health officials with critical insights into the progression, control, and prevention of disease spread. Of these models, one of most fundamental is the SIR differential equation model. However, this ubiquitous model has one significant and rarely acknowledged shortcoming: it is unable to account for a diseases true infectious period distribution. As the misspecification of such a biological characteristic is known to significantly affect model behavior, there is a need to develop new modeling approaches that capture such information. Therefore, we illustrate an innovative take on compartmental models, derived from their general formulation as systems of nonlinear Volterra integral equations, to capture a broader range of infectious period distributions, yet maintain the desirable formulation as systems of differential equations. Our work illustrates a compartmental model that captures any Erlang distributed duration of infection with only 3 differential equations, instead of the typical inflated model sizes required by differential equation compartmental models, and a compartmental model that capture any mean, standard deviation, skewness, and kurtosis of an infectious period distribution with merely 4 differential equations. The significance of our work is that it opens up a new class of easy-to-use compartmental models to predict disease outbreaks that does not require a complete overhaul of existing theory, and thus provides a starting point for multiple research avenues of investigation under the contexts of mathematics, public health, and evolutionary biology.

microbiology↗

The Genome of the Wasp Anastatus disparis Reveals Energy Metabolism Adaptations for Extreme Aggression.

Extremely aggressive behavior is rare in most species, as contestants can be severely injured or killed. Such high level of aggression can evolve when critical resources are limited, as the benefits of winning outweigh the potential costs of conflict. Currently, studies of extreme aggression are mainly from the perspectives of behavioral ecology and evolutionary biology, displaying distinct results from common nonextreme aggression. Here, we provide a high-quality genome of the generalist endoparasitoid Anastatus disparis, the males of which exhibit extreme mate-competition aggression, to shed light on possible genomic adaptations for extreme aggressive behavior. We combined Nanopore PromethION sequencing with Hi-C technology to assemble a high-quality chromosome-level genome of A. disparis. The size of the genome of this species (939.58 Mb) is larger than that of most hymenopterans (ranging from 180 Mb to 340 Mb) due to the expansion of repeated sequences (612.90 Mb, 65.23% of the whole genome). With the aid of RNA sequencing, 19,246 protein-coding genes were identified, and a great expansion of genes involved in detoxification was detected, which could represent an adaptation of this species to exploit a diverse range of known hosts. The integrated multiomics analysis highlighted genes involved in energy metabolism (especially from lipids) and antibacterial activity, both of which are possibly major aspects of adaptation for extreme aggression in A. disparis. Our study provides insight into molecular and evolutionary studies of extreme aggression in A. disparis and provides a valuable genomic resource for further research into the molecular basis of trait evolution in Hymenoptera.

genomics↗

Perspective: Genomic inference using diffusion models and the allele frequency spectrum

Evolutionary, biological, and demographic processes combine to shape the variation observed in populations. Understanding how these processes are expected to influence variation allows us to infer past demographic events and the nature of selection in human populations. Forward models such as the diffusion approximation provide a powerful tool for analyzing the distribution of allele frequencies in contemporary populations due to their computational tractability and model flexibility. Here, we discuss recent computational developments and their application to reconstructing human demographic history and patterns of selection at new mutations. We also reexamine how some classical assumptions that are still commonly used in inference studies fare when applied to modern data. We use whole-genome sequence data for 797 French Canadian individuals to examine the neutrality of synonymous sites. We find that selection can lead to strong biases in the inferred demography, mutation rate, and distributions of fitness effects. We use these distributions of fitness effects together with demographic and phenotype-fitness models to predict the relationship between effect size and allele frequency, and contrast those predictions to commonly used models in statistical genetics. Thus the simple evolutionary models investigated by Kimura and Ohta still provide important insight into modern genetic research.

genetics↗

Chlomito: a novel tool for precise elimination of organelle genome contamination in nuclear genome assemblies

Accurate genome assemblies are crucial for understanding biological evolution, mechanisms of disease, and biodiversity. However, contamination from organelle genomes in nuclear genome analyses often leads to inaccuracies and unreliability in results. To address this issue, we developed a tool named Chlomito, which employs innovative algorithms to precisely identify and eliminate organelle genome contamination sequences from nuclear genome assemblies. Compared to conventional approaches, Chlomito can not only detect and eliminate organelle sequences but also effectively distinguish true organelle sequences from those transferred into the nucleus via horizontal gene transfer. To evaluate the accuracy of Chlomito, we conducted tests using sequencing data from Plum and Mango. The results confirmed that Chlomito can accurately detect contigs originating from the organelle genome, and the identified contigs covered most regions of the organelle reference genomes, demonstrating its efficiency and precision in comprehensively recognizing organelle genome sequences. Additionally, for user convenience, we packaged this method into a Docker image, simplifying the data processing workflow. Overall, Chlomito provides a highly efficient and accurate method for identifying and removing contigs derived from organelle genomes in genomic assembly data, thereby contributing to the improvement of genome assembly quality and advancing research in genomics and evolutionary biology.

bioinformatics↗

A global perspective of codon usage

Codon usage in 2730 genomes is analyzed for evolutionary patterns in the usage of synonymous codons and amino acids across prokaryotic and eukaryotic taxa. We group genomes together that have similar amounts of intra-genomic bias in their codon usage, and then compare how usage of particular different codons is diversified across each genome group, and how that usage varies from group to group. Inter-genomic diversity of codon usage increases with intra-genomic usage bias, following a universal pattern. The frequencies of the different codons vary in robust mutual correlation, and the implied synonymous codon and amino acid usages drift together. This kind of correlation indicates that the variation of codon usage across organisms is chiefly a consequence of lateral DNA transfer among diverse organisms. The group of genomes with the greatest intra-genomic bias comprises two distinct subgroups, with each one restricting its codon usage to essentially one unique half of the genetic code table. These organisms include eubacteria and archaea thought to be closest to the hypothesized last universal common ancestor (LUCA). Their codon usages imply genetic diversity near the hypothesized base of the tree of life. There is a continuous evolutionary progression across taxa from the two extremely diversified usages toward balanced usage of different codons (as approached, e.g. in mammals). In that progression, codon frequency variations are correlated as expected from a blending of the two extreme codon usages seen in prokaryotes.\n\nAUTHOR SUMMARYThe redundancy intrinsic to the genetic code allows different amino acids to be encoded by up to six synonymous codons. Genomes of different organisms prefer different synonymous codons, a phenomenon known as codon usage bias. The phenomenon of codon usage bias is of fundamental interest for evolutionary biology, and is important in a variety of applied settings (e.g., transgene expression). The spectrum of codon usage biases seen in current organisms is commonly thought to have arisen by the combined actions of mutations and selective pressures. This view focuses on codon usage in specific genomes and the consequences of that usage for protein expression.\n\nHere we investigate an unresolved question of molecular genetics: are there global rules governing the usage of synonymous codons made by genomic DNA across organisms? To answer this question, we employed a data-driven approach to surveying 2730 species from all kingdoms of the tree of life in order to classify their codon usage. A first major result was that the large majority of these organisms use codons rather uniformly on the genome-wide scale, without giving preference to particular codons among possible synonymous alternatives. A second major result was that two compartments of codon usage seem to co-exist and to be expressed in different proportions by different organisms. As such, we investigate how individual different codons are used in different organisms from all taxa. Whereas codon usage is generally believed to be the evolutionary result of both mutations and natural selection, our results suggest a different perspective: the usage of different codons (and amino acids) by different organisms follows a superposition of two distinct patterns of usage. One distinction locates to the third base pair of all different codons, which in one pattern is U or A, and in the other pattern is G or C. This result has two major implications: (1) the variation of codon usage as seen across different organisms is best accounted for by lateral gene transfer among diverse organisms; (2) the organisms that are by protein homology grouped near the base of the tree of life comprise two genetically distinct lineages.\n\nWe find that, over evolutionary time, codon usages have converged from two distinct, non-overlapping usages (e.g., as evident in bacteria and archaea) to a near-uniform, balanced usage of synonymous codons (e.g., in mammals). This shows that the variations of codon (and amino acid) biases reveal a distinct evolutionary progression. We also find that codon usage in bacteria and archaea is most diverse between organisms thought to be closest to the hypothesized last universal common ancestor (LUCA). The dichotomy in codon (and amino acid usages) present near the origin of the current tree of life might provide information about the evolutionary development of the genetic code.

Genetics↗

Dispersal ability predicts evolutionary success among mammalian carnivores

Understanding why some clades contain more species than others is a major challenge in evolutionary biology, and variation in dispersal ability and its connection to diversification rate may be part of the explanation. Several studies have suggested a negative relationship between dispersal capacity and diversification rate among living mammals. However, this pattern may differ when also considering extinct species, given known extinction biases. The colonization of new areas by various lineages may be associated with both diversity increases in those colonising lineages and declines in the lineages already present. Past diversity declines are, however, effectively impossible to infer based on phylogenies of extant taxa, and the underlying process may, therefore, be difficult to determine. Here we produce a novel species-level phylogeny of all known extant and extinct species of the order Carnivora and related extinct groups (1,723 species in total) to show that there is instead a positive relationship between dispersal rate and diversification rate when all extinct species are included. Species that disperse between continents leave more descendant species than non-dispersers, and dispersing species belong to lineages that at the time of dispersal were diversifying faster than the average non-disperser. Our study showcases the importance of combining fossils and phylogenies to better understand evolutionary and biogeographic patterns.View Full Text

paleontology↗

A new high-quality genome assembly and annotation for the threatened Florida Scrub-Jay (Aphelocoma coerulescens)

The Florida Scrub-Jay (Aphelocoma coerulescens), a Federally Threatened, cooperatively-breeding bird, is an emerging model system in evolutionary biology and ecology. Extensive individual-based monitoring and genetic sampling for decades has yielded a wealth of data, allowing for the detailed study of social behavior, demography, and population genetics of this natural population. Here, we report a linkage map and a chromosome-level genome assembly and annotation for a female Florida Scrub-Jay made with long-read sequencing technology, chromatin conformation data, and the linkage map. We constructed a linkage map comprising 4,468 SNPs that had 34 linkage groups and a total sex-averaged autosomal genetic map length of 2446.78 cM. The new genome assembly is 1.32 Gb in length, consisting of 33 complete or near-complete autosomes and the sex chromosomes (ZW). This highly contiguous assembly has an N50 of 68 Mb and a Benchmarking Universal Single-Copy Orthologs (BUSCO) completeness score of 97.1% with respect to the Aves database. The annotated gene set has a BUSCO transcriptome completeness score of 95.5% and 18,051 identified protein-coding genes, 92.2% of which have associated functional annotations. This new, high-quality genome assembly and linkage map of the Florida Scrub-Jay provides valuable tools for future research into the evolutionary dynamics of small, natural populations of conservation concern. ARTICLE SUMMARYWe present a new high-quality genome assembly and annotation for the Florida Scrub-Jay (Aphelocoma coerulescens), a Federally Threatened bird species. In comparison to other genome assemblies of this species, our assembly is the first to be made using long-read sequencing technology and is the first generated from a female individual. We also constructed the first linkage map for this species using a population pedigree. Our genome assembly is highly contiguous and is of similar quality to other bird genome assemblies.

genomics↗

ngsAMOVA: A Probabilistic Framework for Analysis of Molecular Variance, dXY and Neighbor-Joining Trees with Low Depth Sequencing Data

MotivationNext-generation sequencing (NGS) has transformed population genetics and evolutionary biology, but the data produced in studies of non-model organisms, ancient DNA, and environmental DNA often consist of low- or medium-depth sequencing. Analyses of these data rely on computational methods that utilize genotype likelihoods (GLs) to account for genotype uncertainty. Nevertheless, many widely-used analysis methods, such as analysis of molecular variance (AMOVA) and methods for estimating phylogenetic trees using nucleotide divergence (dXY) still lack the probabilistic frameworks necessary to accommodate GLs. ResultsWe introduce ngsAMOVA, a novel probabilistic framework for analyzing molecular variation in population hierarchies with low- and medium-depth sequencing data. It employs an Expectation Maximization algorithm to first estimate the joint genotype probabilities for pairs of individuals, accounting for genotype uncertainty using GLs. It then uses these estimates to generate a pairwise distance matrix, which can be used for AMOVA, estimation of dXY, and for estimating phylogenetic trees using Neighbor-Joining. Hypothesis testing is facilitated using genomic block-bootstrapping. Through extensive simulations, we demonstrate that ngsAMOVA provides more accurate results compared to genotype calling at low and medium read depths. Overall, ngsAMOVA represents a methodological advance in the analysis of molecular variance and divergence under sequencing uncertainty. It provides a robust framework, opening up numerous possibilities for gaining insights into the evolutionary histories through its applications. ngsAMOVA is available as a fast, efficient, and user-friendly program written in C/C++. AvailabilityngsAMOVA is freely available at https://github.com/isinaltinkaya/ngsAMOVA. Contactisin.altinkaya@sund.ku.dk Supplementary informationSupplementary data are available online.

bioinformatics↗

Taking advantage of reference-guided assembly in a slowly-evolving lineage: application to Testudo graeca.

BackgroundObtaining de novo chromosome-level genome assemblies greatly enhances conservation and evolutionary biology studies. For many research teams, long-read sequencing technologies (that produce highly contiguous assemblies) remain unaffordable or unpractical. For the groups that display high synteny conservation, these limitations can be overcome by a reference-guided assembly using a close relative genome. Of chelonians, terrestrial tortoises are considered one of the most endangered taxa, which calls for more genomic resources. Here we make the most of high synteny conservation in chelonians to produce the first chromosome-level genome assembly of genus Testudo with one of the most iconic tortoise species in the Mediterranean basin: T. graeca. ResultsWe used high quality, paired-end Illumina sequences to build a reference-guided assembly with the chromosome level assembly of Gopherus evgoodei. We reconstructed a 2.29 Gb haploid genome with a scaffold N50 of 107.598 Mb and 5.37% gaps. We sequenced 25998 protein-coding genes, and a 41.2% fraction was determined as repetitive in our assembled genome. Demographic history reconstruction based on the genome revealed two events (population decline and recovery) consistent with previously suggested phylogeographic patterns for the species. This outlines the value of genomes like this for phylogeographic studies. ConclusionsOur results highlight the value of using close relatives to produce de novo draft assemblies in species where such resources are unavailable. Our Testudo graeca annotated genome paves the way to delve deeper into the species evolutionary history and provides a valuable resource to increase making direct conservation efforts on their threatened populations.

genomics↗

Cytogenetic constraints on hybridization: A meta-analysis investigating the role of chromosome number in monocot hybrid evolution using a newly developed tool, the ploidy deviation index (PDI).

Background and aimsHybridization is a major driver of plant diversity, yet the role of cytogenetic compatibility, particularly differences in chromosome number, remains poorly understood. Differences in parental chromosome number can present barriers to hybrid formation by disrupting meiotic stability, but the extent to which biological and ecological factors influence the chromosomal architecture of hybrids remains poorly quantified, especially in monocots. This study aims to investigate how chromosome number divergence interacts with biological and ecological factors to shape hybrid formation in monocots, using a novel quantitative metric, the Ploidy Deviation Index (PDI), to standardize comparisons of hybrid cytogenetic architecture. Material and methodsWe developed and applied the PDI, a continuous index quantifying chromosome-number deviation between a hybrid and its two parents, across approximately 200 hybrid cases with documented parental karyotypes. Hybrids were categorized as homoploid, uniparentally homoploid, intermediate, or polyploid based on their PDI values. We analyzed the distribution of PDI scores in relation to type of hybrid origin (natural vs. artificial), growth habit, size of the genus (a proxy for richness), and range of chromosome number within a genus (proxy for diversity). Comparisons across categories employed Anderson-Darling k-sample tests, multinomial logistic regression, and Mann-Whitney U tests to determine significance. Key resultsHomoploid hybrids were found to be the most frequent. We found no significant difference in PDI distributions between natural and artificial hybrids. Significant variation in PDI distributions was found among growth habits, with aquatic hybrids more likely to be homoploid and geophytic hybrids showing higher proportions of polyploidy. Intermediate hybrids were common in larger genera with broader chromosome-number ranges, whereas polyploid hybrids showed the highest PDI values in large and karyotypically diverse genera. ConclusionThese results challenge long-held assumptions that polyploidy dominates hybrid formation and reveal that homoploid and intermediate chromosomal configurations are common in monocots. The PDI framework offers a powerful, standardized approach for assessing cytogenetic constraints on hybridization, with implications for systematics, evolutionary biology, and conservation.

plant biology↗