Noland H. Martin

Aim: This study sought to evaluate how historical drainage connectivity and contemporary basin isolation have shaped population genetic structure and diversity in an imperilled species of freshwater mussel, Pleurobema riddellii. Location: Covers the range of P. riddellii , spanning seven river basins of the Gulf Coastal Plains of the southern USA, including the San Jacinto, Neches, Sabine, Calcasieu, Red, Ouachita and Pearl river systems. Methods: A genome‐wide single nucleotide polymorphism (SNP) dataset was generated using a restriction‐site associated DNA sequencing approach. After filtering and assembly, a dataset consisting of 12,062 SNP loci from 351 individuals was retained. Population genetic structure was evaluated using principal components analysis and a Bayesian clustering model, ENTROPY. Genetic differentiation among river basins was quantified using Nei's G _ST and patterns of genetic diversity were assessed using observed heterozygosity and Watterson's θ . Results: Population genetic analyses revealed structuring that corresponded closely with river basin boundaries. Centrally distributed populations from the Neches, Sabine and Calcasieu basins formed a genetically cohesive group characterised by low genetic differentiation and elevated genetic diversity. Marginal populations from the San Jacinto, Red, Ouachita and Pearl basins exhibited higher levels of genetic differentiation and reduced genetic diversity. Main Conclusions: Here, we demonstrate a central‐marginal pattern in genomic structure and diversity across the range of P. riddellii . These results suggest that historical hydrological connectivity, coupled with prolonged isolation and reduced effective population sizes at range margins, likely played a role in shaping standing genomic variation in this system. More broadly, the genomic structure observed in P. riddellii is consistent with other unionid mussels in the region, highlighting the contributions of historical hydrological configurations in structuring freshwater biodiversity.

The evolution of reproductive isolation (RI) results in the reduction of interspecific hybridization and the maintenance of species boundaries. Asymmetries in RI, where one species more frequently serves as the maternal or paternal parent in initial F₁ hybrid formation, are commonly observed in plants. Asymmetric introgression, the predominantly unidirectional transfer of genetic material through hybridization and backcrossing, has also been frequently documented in hybridizing plant taxa as well. This study investigates whether asymmetries in total RI measured between species can predict the direction of introgression in naturally hybridizing plant taxa. A meta-analysis was conducted on 19 plant species pairs with published data on both asymmetric total RI, and asymmetric introgression. Species pairs that met these criteria were identified through a comprehensive literature review. A two-tailed binomial test was performed to evaluate whether asymmetric RI was associated with asymmetries in introgression. No significant relationship was found between asymmetries in total RI and the direction of introgression (p = 0.3593). Asymmetric RI largely does not predict the direction of introgression. Rather, introgression patterns may be better understood by examining F₁ and later-generation hybrids in natural settings, focusing on their fitness, mating behaviors, and the ecological and demographic factors that shape hybrid zones.

Background/Objectives: The evolution of reproductive isolation (RI) results in the reduction of interspecific hybridization and the maintenance of species boundaries. Asymmetries in RI, where one species more frequently serves as the maternal or paternal parent in initial F1 hybrid for-mation, are commonly observed in plants. Asymmetric introgression, the predominantly unidi-rectional transfer of genetic material through hybridization and backcrossing, has also been fre-quently documented in hybridizing plant taxa as well. This study investigates whether asymme-tries in total RI measured between species can predict the direction of introgression in naturally hybridizing plant taxa. Methods: A meta-analysis was conducted on 19 plant species pairs with published data on both asymmetric total RI, and asymmetric introgression. Species pairs that met these criteria were identified through a comprehensive literature review. A two-tailed binomial test was performed to evaluate whether asymmetric RI was associated with asymmetries in in-trogression. Results: No significant relationship was found between asymmetries in total RI and the direction of introgression (p = 0.3593). Conclusions: Asymmetric RI largely does not predict the direction of introgression. Rather, introgression patterns may be better understood by exam-ining F1 and later-generation hybrids in natural settings, focusing on their fitness, mating behav-iors, and the ecological and demographic factors that shape hybrid zones.

Limiting harm to organisms caused by genetic sampling is an important consideration for rare species, and a number of non-destructive sampling techniques have been developed to address this issue in freshwater mussels. Two methods, visceral swabbing and tissue biopsies, have proven to be effective for DNA sampling, though it is unclear as to which method is preferable for genotyping-by-sequencing (GBS). Tissue biopsies may cause undue stress and damage to organisms, while visceral swabbing potentially reduces the chance of such harm. Our study compared the efficacy of these two DNA sampling methods for generating GBS data for the unionid freshwater mussel, the Texas pigtoe (Fusconaia askewi). Our results find both methods generate quality sequence data, though some considerations are in order. Tissue biopsies produced significantly higher DNA concentrations and larger numbers of reads when compared with swabs, though there was no significant association between starting DNA concentration and number of reads generated. Swabbing produced greater sequence depth (more reads per sequence), while tissue biopsies revealed greater coverage across the genome (at lower sequence depth). Patterns of genomic variation as characterized in principal component analyses were similar regardless of the sampling method, suggesting that the less invasive swabbing is a viable option for producing quality GBS data in these organisms.

Premise: When divergent lineages come into secondary contact, reproductive isolation may be incomplete, thus providing an opportunity to investigate how speciation is manifested in the genome. The Louisiana Irises (Iris, series Hexagonae) comprise a group of three or more ecologically and reproductively divergent lineages that can produce hybrids where they come into contact. We estimated standing genetic variation to understand the current distribution of population structure in the Louisiana Irises.Methods: We used genotyping‐by‐sequencing techniques to sample the genomes of Louisiana Iris species across their ranges. We sampled 20 populations (n = 632 individuals) across 11,249 loci and used Entropy and PCA models to assess population genetic data.Results: We discovered evidence for interspecific gene flow in parts of the range. Our analysis revealed patterns of population structure at odds with widely accepted nominal taxonomy. We discovered undescribed hybrid populations, designated as belonging to the I. brevicaulis lineage. Iris nelsonii shared significant ancestry with only one of the purported parent species, I. fulva, evidence inconsistent with a hybrid origin.Conclusions: This study provides several key findings important to the investigation of standing genetic variation in the Louisiana Iris species complex. Compared to the other nominal species, I. brevicaulis contains a large amount of genetic diversity. In addition, we discovered a previously unknown hybrid zone between I. brevicaulis and I. hexagona along the Texas coast. Finally, our results do not support the long‐standing hypothesis that I. nelsonii has mixed ancestry from three parental taxa.

When ecologically divergent taxa encounter one another, hybrid zones can form when reproductive isolation is incomplete. The location of such hybrid zones can be influenced by environmental variables, and an ecological context can provide unique insights into the mechanisms by which species diverge and are maintained. Two ecologically differentiated species of small benthic fishes, the endemic and imperiled prairie chub,Macrhybopsis australis, and the shoal chub,Macrhybopsis hyostoma, are locally sympatric within the upper Red River Basin of Texas. We integrated population genomic data and environmental data to investigate species divergence and the maintenance of species boundaries in these two species. We found evidence of advanced‐generation asymmetric hybridization and introgression, with shoal chub alleles introgressing more frequently into prairie chubs than the reciprocal. Using a Bayesian Genomic Cline framework, patterns of genomic introgression were revealed to be quite heterogeneous, yet shoal chub alleles were found to have likely selectively introgressed across species boundaries significantly more often than prairie chub alleles, potentially explaining some of the observed asymmetry in hybridization. These patterns were remarkably consistent across two sampled geographic regions of hybridization. Several environmental variables were found to significantly predict individual admixture, suggesting ecological isolation might maintain species boundaries.

Hybrid zones provide unique opportunities to examine reproductive isolation and introgression in nature. We utilized 45,384 single nucleotide polymorphism (SNP) loci to perform association mapping of 14 floral, vegetative and ecological traits that differ betweenIris hexagonaandIris fulva,and to investigate, using a Bayesian genomic cline (BGC) framework, patterns of genomic introgression in a large and phenotypically diverse hybrid zone in southern Louisiana. Many loci of small effect size were consistently found to be associated with phenotypic variation across all traits, and several individual loci were revealed to influence phenotypic variation across multiple traits. Patterns of genomic introgression were quite heterogeneous throughout the Louisiana Iris genome, withI. hexagonaalleles tending to be favoured over those ofI. fulva. Loci that were found to have exceptional patterns of introgression were also found to be significantly associated with phenotypic variation in a small number of morphological traits. However, this was the exception rather than the rule, as most loci that were associated with morphological trait variation were not significantly associated with excess ancestry. These findings provide insights into the complexity of the genomic architecture of phenotypic differences and are a first step towards identifying loci that are associated with both trait variation and reproductive isolation in nature.

Background: Hybridization among Louisiana Irises has been well established and the genetic architecture of reproductive isolation is known to affect the potential for and the directionality of introgression between taxa. Here we use co-dominant markers to identify regions where QTL are located both within and between backcross maps to compare the genetic architecture of reproductive isolation and fitness traits across treatments and years. Results: QTL mapping was used to elucidate the genetic architecture of reproductive isolation between Iris fulva and Iris brevicaulis. Homologous co-dominant EST-SSR markers scored in two backcross populations between I. fulva and I. brevicaulis were used to generate genetic linkage maps. These were used as the framework for mapping QTL associated with variation in 11 phenotypic traits likely responsible for reproductive isolation and fitness. QTL were dispersed throughout the genome, with the exception of one region of a single linkage group (LG) where QTL for flowering time, sterility, and fruit production clustered. In most cases, homologous QTL were not identified in both backcross populations, however, homologous QTL for flowering time, number of growth points per rhizome, number of nodes per inflorescence, and number of flowers per node were identified on several linkage groups. Conclusions: Two different traits affecting reproductive isolation, flowering time and sterility, exhibit different genetic architectures, with numerous QTL across the Iris genome controlling flowering time and fewer, less distributed QTL affecting sterility. QTL for traits affecting fitness are largely distributed across the genome with occasional overlap, especially on LG 4, where several QTL increasing fitness and decreasing sterility cluster. Given the distribution and effect direction of QTL affecting reproductive isolation and fitness, we have predicted genomic regions where introgression may be more likely to occur (those regions associated with an increase in fitness and unlinked to loci controlling reproductive isolation) and those that are less likely to exhibit introgression (those regions linked to traits decreasing fitness and reproductive isolation).

The aim of this study is to investigate the evolution of intrinsic postzygotic isolation within and between populations of Mimulus guttatus and Mimulus nasutus . We made 17 intraspecific and interspecific crosses, across a wide geographical scale. We examined the seed germination success and pollen fertility of reciprocal F ₁ and F ₂ hybrids and their pure-species parents, and used biometrical genetic tests to distinguish among alternative models of inheritance. Hybrid seed inviability was sporadic in both interspecific and intraspecific crosses. For several crosses, Dobzhansky–Muller incompatibilities involving nuclear genes were implicated, while two interspecific crosses revealed evidence of cytonuclear interactions. Reduced hybrid pollen fertility was found to be greatly influenced by Dobzhansky–Muller incompatibilities in five out of six intraspecific crosses and nine out of 11 interspecific crosses. Cytonuclear incompatibilities reduced hybrid fitness in only one intraspecific and one interspecific cross. This study suggests that intrinsic postzygotic isolation is common in hybrids between these Mimulus species, yet the particular hybrid incompatibilities responsible for effecting this isolation differ among the populations tested. Hence, we conclude that they evolve and spread only at the local scale.