Hybrid Zone Research Articles

Mitonuclear interactions impact aerobic metabolism in hybrids and may explain mitonuclear discordance in young, naturally hybridizing bird lineages.

Understanding genetic incompatibilities and genetic introgression between incipient species are major goals in evolutionary biology. Mitochondrial genes evolve rapidly and exist in dense gene networks with coevolved nuclear genes, suggesting that mitochondrial respiration may be particularly susceptible to disruption in hybrid organisms. Mitonuclear interactions have been demonstrated to contribute to hybrid dysfunction between deeply divergent taxa crossed in the laboratory, but there are few empirical examples of mitonuclear interactions between younger lineages that naturally hybridize. Here, we use controlled hybrid crosses and high-resolution respirometry to provide the first experimental evidence in a bird that inter-lineage mitonuclear interactions impact mitochondrial aerobic metabolism. Specifically, respiration capacity of the two mitodiscordant backcrosses (with mismatched mitonuclear combinations) differs from one another, although they do not differ significantly from the parental groups or mitoconcordant backcrosses as we would expect of mitonuclear disruptions. In the wild hybrid zone between these subspecies, the mitochondrial cline centre is shifted west of the nuclear cline centre, which is consistent with the direction of our experimental results. Our results therefore demonstrate asymmetric mitonuclear interactions that impact the capacity of cellular mitochondrial respiration and may help to explain the geographic discordance between mitochondrial and nuclear genomes observed in the wild.

Selection leads to remarkable variability in the outcomes of hybridisation across replicate hybrid zones.

Hybrid zones have been viewed as an opportunity to see speciation in action. When hybrid zones are replicated, it is assumed that if the same genetic incompatibilities are maintaining reproductive isolation across all instances of secondary contact, those incompatibilities should be identifiable by consistent patterns in the genome. In contrast, changes in allele frequencies due to genetic drift should be idiosyncratic for each hybrid zone. To test this assumption, we simulated 20 replicates of each of 12 hybrid zone scenarios with varied genetic incompatibilities, rates of migration, selection and different starting population size ratios of parental species. We found remarkable variability in the outcomes of hybridisation in replicate hybrid zones, particularly with Bateson-Dobzhansky-Muller incompatibilities and strong selection. We found substantial differences among replicates in the overall genomic composition of individuals, including admixture proportions, inter-specific ancestry complement and number of ancestry junctions. Additionally, we found substantial variation in genomic clines among replicates at focal loci, regardless of locus-specific selection. We conclude that processes other than selection are responsible for some consistent outcomes of hybridisation, whereas selection on incompatibilities can lead to genomically widespread and highly variable outcomes. We highlight the challenge of mapping between pattern and process in hybrid zones and call attention to how selection against incompatibilities will commonly lead to variable outcomes. We hope that this study informs future research on replicate hybrid zones and encourages further development of statistical techniques, theoretical models and exploration of additional axes of variation to understand reproductive isolation.

Association between the gut microbiome and carotenoid plumage phenotype in an avian hybrid zone.

Vertebrates host complex microbiomes that impact their physiology. In many taxa, including colourful wood-warblers, gut microbiome similarity decreases with evolutionary distance. This may suggest that as host populations diverge, so do their microbiomes, because of either tight coevolutionary dynamics, or differential environmental influences, or both. Hybridization is common in wood-warblers, but the effects of evolutionary divergence on the microbiome during secondary contact are unclear. Here, we analyse gut microbiomes in two geographically disjunct hybrid zones between blue-winged warblers (Vermivora cyanoptera) and golden-winged warblers (Vermivora chrysoptera). We performed 16S faecal metabarcoding to identify species-specific bacteria and test the hypothesis that host admixture is associated with gut microbiome disruption. Species identity explained a small amount of variation between microbiomes in only one hybrid zone. Co-occurrence of species-specific bacteria was rare for admixed individuals, yet microbiome richness was similar among admixed and parental individuals. Unexpectedly, we found several bacteria that were more abundant among admixed individuals with a broader deposition of carotenoid-based plumage pigments. These bacteria are predicted to encode carotenoid biosynthesis genes, suggesting birds may take advantage of pigments produced by their gut microbiomes. Thus, host admixture may facilitate beneficial symbiotic interactions which contribute to plumage ornaments that function in sexual selection.

Italian peninsula as a hybridization zone of Ixodes inopinatus and I. ricinus and the prevalence of tick-borne pathogens in I. inopinatus, I. ricinus, and their hybrids

BackgroundIxodes inopinatus was described from Spain on the basis of morphology and partial sequencing of 16S ribosomal DNA. However, several studies suggested that morphological differences between I. inopinatus and Ixodes ricinus are minimal and that 16S rDNA lacks the power to distinguish the two species. Furthermore, nuclear and mitochondrial markers indicated evidence of hybridization between I. inopinatus and I. ricinus. In this study, we tested our hypothesis on tick dispersal from North Africa to Southern Europe and determined the prevalence of selected tick-borne pathogens (TBPs) in I. inopinatus, I. ricinus, and their hybrids.MethodsTicks were collected in Italy and Algeria by flagging, identified by sequencing of partial TROSPA and COI genes, and screened for Borrelia burgdorferi s.l., B. miyamotoi, Rickettsia spp., and Anaplasma phagocytophilum by polymerase chain reaction and sequencing of specific markers.ResultsOut of the 380 ticks, in Italy, 92 were I. ricinus, 3 were I. inopinatus, and 136 were hybrids of the two species. All 149 ticks from Algeria were I. inopinatus. Overall, 60% of ticks were positive for at least one TBP. Borrelia burgdorferi s.l. was detected in 19.5% of ticks, and it was significantly more prevalent in Ixodes ticks from Algeria than in ticks from Italy. Prevalence of Rickettsia spotted fever group (SFG) was 51.1%, with significantly greater prevalence in ticks from Algeria than in ticks from Italy. Borrelia miyamotoi and A. phagocytophilum were detected in low prevalence (0.9% and 5.2%, respectively) and only in ticks from Italy.ConclusionsThis study indicates that I. inopinatus is a dominant species in Algeria, while I. ricinus and hybrids were common in Italy. The higher prevalence of B. burgdorferi s.l. and Rickettsia SFG in I. inopinatus compared with that in I. ricinus might be due to geographical and ecological differences between these two tick species. The role of I. inopinatus in the epidemiology of TBPs needs further investigation in the Mediterranean Basin.Graphical

Tufted titmice, black-crested titmice, and their hybrids occupy different types of habitat within their hybrid zone

Tufted titmice, black-crested titmice, and their hybrids occupy different types of habitat within their hybrid zone

Phenotypic selection patterns in a hybrid zone between two Calceolaria species with contrasting pollinators: insights from field surveys and fitness assessments.

Hybrid zones provide natural experimental settings to test hypotheses about species divergence. We concentrated on a hybrid swarm in which oil-collecting bees and flower-pecking birds act as pollinators of two Calceolaria species. We asked whether both pollinators contributed to flower divergence by differentially promoting prezygotic fitness at the phenotypic extremes that represent parentals. We studied pollinator-mediated selection on phenotypic traits critical in plant-pollinator mechanical interaction, namely plant height, reward-to-stigma distance, and flower shape. We utilised the quantity and quality of pollen deposited as fitness measures and distinguished between the contribution of the two pollinator types. Results showed uni- and bivariate disruptive selection for most traits through pollen grains deposited by both pollinators. Bird-mediated fitness favoured low plants with a long reward-to-stigma distance and a straight corolla, while bee-mediated fitness favoured tall plants with a short reward-to-stigma distance and curved corolla. In addition, stabilising selection at one end of the phenotypic range showed a bird-mediated reproductive asymmetry within the swarm. The disruptive pattern was countered, albeit weakly, by hybrids receiving higher-quality pollen on the stigmas. Results suggest that pollinator-mediated selection promotes divergence of integrated flower phenotypes mechanically adjusted either to bees or birds underscoring the importance of pollinator specialisation in diversification.

Coupling of twelve putative chromosomal inversions maintains a strong barrier to gene flow between snail ecotypes

Abstract Chromosomal rearrangements can lead to the coupling of reproductive barriers, but whether and how they contribute to the completion of speciation remains unclear. Marine snails of the genus Littorina repeatedly form hybrid zones between populations segregating for multiple inversion arrangements, providing opportunities to study their barrier effects. Here, we analyzed 2 adjacent transects across hybrid zones between 2 ecotypes of Littorina fabalis (“large” and “dwarf”) adapted to different wave exposure conditions on a Swedish island. Applying whole-genome sequencing, we found 12 putative inversions on 9 of 17 chromosomes. Nine of the putative inversions reached near differential fixation between the 2 ecotypes, and all were in strong linkage disequilibrium. These inversions cover 20% of the genome and carry 93% of divergent single nucleotide polymorphisms (SNPs). Bimodal hybrid zones in both transects indicated that the 2 ecotypes of Littorina fabalis maintain their genetic and phenotypic integrity following contact. The bimodality reflects the strong coupling between inversion clines and the extension of the barrier effect across the whole genome. Demographic inference suggests that coupling arose during a period of allopatry and has been maintained for > 1,000 generations after secondary contact. Overall, this study shows that the coupling of multiple chromosomal inversions contributes to strong reproductive isolation. Notably, 2 of the putative inversions overlap with inverted genomic regions associated with ecotype differences in a closely related species (Littorina saxatilis), suggesting the same regions, with similar structural variants, repeatedly contribute to ecotype evolution in distinct species.

A genomic basis of vocal rhythm in birds

Vocal rhythm plays a fundamental role in sexual selection and species recognition in birds, but little is known of its genetic basis due to the confounding effect of vocal learning in model systems. Uncovering its genetic basis could facilitate identifying genes potentially important in speciation. Here we investigate the genomic underpinnings of rhythm in vocal non-learning Pogoniulus tinkerbirds using 135 individual whole genomes distributed across a southern African hybrid zone. We find rhythm speed is associated with two genes that are also known to affect human speech, Neurexin-1 and Coenzyme Q8A. Models leveraging ancestry reveal these candidate loci also impact rhythmic stability, a trait linked with motor performance which is an indicator of quality. Character displacement in rhythmic stability suggests possible reinforcement against hybridization, supported by evidence of asymmetric assortative mating in the species producing faster, more stable rhythms. Because rhythm is omnipresent in animal communication, candidate genes identified here may shape vocal rhythm across birds and other vertebrates.

Mapping seasonal migration in a songbird hybrid zone -- heritability, genetic correlations, and genomic patterns linked to speciation

Seasonal migration is a widespread behavior relevant for adaptation and speciation, yet knowledge of its genetic basis is limited. We leveraged advances in tracking and sequencing technologies to bridge this gap in a well-characterized hybrid zone between songbirds that differ in migratory behavior. Migration requires the coordinated action of many traits, including orientation, timing, and wing morphology. We used genetic mapping to show these traits are highly heritable and genetically correlated, explaining how migration has evolved so rapidly in the past and suggesting future responses to climate change may be possible. Many of these traits mapped to the same genomic regions and small structural variants indicating the same, or tightly linked, genes underlie them. Analyses integrating transcriptomic data indicate cholinergic receptors could control multiple traits. Furthermore, analyses integrating genomic differentiation further suggested genes underlying migratory traits help maintain reproductive isolation in this hybrid zone.

Hybrid Fabrication of Cold Metal Transfer Additive Manufacturing and Laser Metal Deposition for Ti6Al4V: The Microstructure and Dynamic/Static Mechanical Properties.

The titanium alloy components utilized in the aviation field are typically large in size and possess complex structures. By utilizing multiple additive manufacturing processes, the precision and efficiency requirements of production can be met. We investigated the hybrid additive manufacturing of Ti-6Al-4V using a combination of cold metal transfer additive manufacturing (CMTAM) and laser metal deposition (LMD), as well as the feasibility of using the CMT-LMD hybrid additive manufacturing process for fabricating Ti-6Al-4V components. Microstructural examinations, tensile testing coupled with digital image correlation and dynamic compressive experiments (by the split Hopkinson pressure bar (SHPB) system) were employed to assess the parts. The results indicate that the interface of the LMD and CMTAM zone formed a compact metallurgical bonding. In the CMTAM and LMD zone, the prior-β grains exhibit epitaxial growth, forming columnar prior-β grains. Due to laser remelting, the CMT-LMD hybrid additive zone experiences grain refinement, resulting in equiaxed prior-β grains at the interface with an average grain size smaller than that of the CMTAM and LMD regions. The microstructures reveal significant differences in grain orientation and morphology among the zones, with distinct textures forming in each zone. In the CMT-LMD hybrid zone, due to interfacial strengthening, strain concentration occurs in the arc additive zone during tensile testing, leading to fracture on the CMTAM zone. Under high-strain-rate dynamic impact conditions, the LMD region exhibits ductile fracture, while the CMTAM zone demonstrates brittle fracture. The hybrid zone combines ductile and brittle fracture modes, and the CMT-LMD hybrid material exhibits superior dynamic impact performance compared to the single deposition zone.

Aberrant microbiomes are associated with increased antibiotic resistance gene load in hybrid mice

Abstract Antibiotic resistance is a priority public health problem resulting from eco-evolutionary dynamics within microbial communities and their interaction at a mammalian host interface or geographical scale. The links between mammalian host genetics, bacterial gut community and antimicrobial resistance gene (ARG) content must be better understood in natural populations inhabiting heterogeneous environments. Hybridisation, the interbreeding of genetically divergent populations, influences different components of the gut microbial communities. However, its impact on bacterial traits such as antibiotic resistance is unknown. Here, we present that hybridisation might shape bacterial communities and ARG occurrence. We used amplicon sequencing to study the gut microbiome and to predict ARG composition in natural populations of house mice (Mus musculus). We compared gastrointestinal bacterial and ARG diversity, composition and abundance across a gradient of pure and hybrid genotypes in the European house mouse hybrid zone. We observed an increased overall predicted richness of ARG in hybrid mice. We found bacteria - ARG interactions by their co-abundance and detected phenotypes of extreme abundances in hybrid mice at the level of specific bacterial taxa and ARGs, mainly multidrug resistance genes. Our work suggests that mammalian host genetic variation impacts the gut microbiome and chromosomal ARGs. However, it raises further questions on how the mammalian host genetics impact ARGs via microbiome dynamics or environmental covariates.

Transgressive gene expression and expression plasticity under thermal stress in a stable hybrid zone.

Interspecific hybridization can lead to myriad outcomes, including transgressive phenotypes in which the hybrids are more fit than either parent species. Such hybrids may display important traits in the context of climate change, able to respond to novel environmental conditions not previously experienced by the parent populations. While this has been evaluated in an agricultural context, the role of transgressive hybrids under changing conditions in the wild remains largely unexplored; this is especially true regarding transgressive gene expression. Using the blue mussel species complex (genus Mytilus) as a model system, we investigated the effects of hybridization on temperature induced gene expression plasticity by comparing expression profiles in parental species and their hybrids following a 2-week thermal challenge. Hybrid expression plasticity was most often like one parent or the other (50%). However, a large fraction of genes (26%) showed transgressive expression plasticity (i.e. the change in gene expression was either greater or lesser than that of both parent species), while only 2% were intermediately plastic in hybrids. Despite their close phylogenetic relationship, there was limited overlap in the differentially expressed genes responding to temperature, indicating interspecific differences in the responses to high temperature in which responses from hybrids are distinct from both parent species. We also identified differentially expressed long non-coding RNAs (lncRNAs), which we suggest may contribute to species-specific differences in thermal tolerance. Our findings provide important insight into the impact of hybridization on gene expression under warming. We propose transgressive hybrids may play an important role in population persistence under future warming conditions.

A taxonomic revision of the Palaearctic species of the ant genus Tapinoma Mayr 1861 (Hymenoptera: Formicidae)

A taxonomic revision of the Palaearctic species of the ant genus Tapinoma Emery 1861 is provided. Due to the extreme rarity of discernable characters, the female castes of 64% of the species are not separable by primary visual perception of an expert and are thus truly cryptic. The main working rationale of the revision is Numeric Morphology-Based Alpha-Taxonomy (NUMOBAT) with formation of species hypotheses largely based on exploratory data analyses and checking these hypotheses by discriminant analysis. NUMOBAT data of all considered species comprise 648 worker samples with 2014 individuals and 30,000 primary data. Species are depicted by z-stack imaging in three standard visual positions. Numeric data on 15 phenotypical characters are presented in comparative tables and supplementary verbal descriptions are given. Analysis of nuclear DNA was based on evaluation of 15 microsatellite loci in 274 nest samples. The NUMOBAT-based classification of eight species of the Tapinoma nigerrimum and T. simrothi group which are inseparable by subjective visual inspection was confirmed by nuclear DNA data in 98.3% of 169 samples where both morphological and nuDNA data were available. The three cases of disagreement between NUMOBAT and nuDNA classification were discussed. Excluding Nomina Nuda and unavailable names, the revision recognized 34 available names which divide into 21 valid species, 11 junior synonyms and two names placed under Incertae Sedis. Four species are described as new: Tapinoma hispanicum n. sp., T. insularis n. sp., T. dabashanica n. sp. and T. sichuense n. sp. The following synonymies were stated: T. collina Foerster 1850, T. erraticum platyops Emery 1925 and T. erraticum bononiensis Emery 1925 as synonyms of T. erraticum (Latreille 1798); T. tauridis Emery 1925 and T. breve Emery 1925 as synonyms of T. glabrella (Nylander 1849) that is erected to species level; T. minor Bernard 1945 as synonym of T. simrothi Krausse-Heldrungen 1911; T. simrothi azerbaidzhanica Karavajev 1932 as synonym of Tapinoma karavaievi Emery 1925 that is erected here to species level; T. erraticum ambiguum Emery 1925 as synonym of T. madeirense Forel 1895 and T. emeryanum Kuznetzov-Ugamsky 1927 as synonym of T. sinense Emery 1925. Despite an insufficient separation of the female castes of Tapinoma madeirense and T. subboreale Seifert 2011 by NUMOBAT characters and detection of a hybrid zone in southern France, the heterospecific status of the two taxa was maintained based on sufficient differences in male genitalia and of nuclear DNA over a larger geographic scale. Formica caerulescens Losana 1834 is removed from genus Tapinoma and recognized as member of the subfamily Formicinae.

A Geometric Morphometrics Approach to the Study of Natural Variations and Hybrid Detection in Populations of Alnus incana (L.) Moench and Alnus rohlenae Vít, Douda and Mandák.

Landmark-based geometric morphometrics (GM) was used to examine, for the first time, spontaneous hybridization between Alnus incana (L.) Moench and Alnus rohlenae Vít, Douda and Mandák, and to assess inter- and intrapopulation variability in leaf shape, leaf size and venation in natural populations in Serbia (Western Balkans). Two geographically distant (30 km) and two close (1.2 km) populations were selected to examine hybridization. The variability in leaf shapes was assessed by canonical variate analysis and linear discriminant analysis performed on the symmetric component of variation. Covariation between the symmetric component of shape variation and the number of pairs of secondary leaf veins was investigated with partial least squares analysis. Static allometry was examined for the first time in the genus Alnus Mill. A higher proportion of A. incana leaves was classified as A. rohlenae in geographically close populations, which is in accordance with the hypothesis about spontaneous hybridization. No single leaf of A. rohlenae was classified as A. incana, indicating that putative hybrids can only be found in grey alder populations. This study demonstrates that GM is a powerful tool for species delimitation and hybrid detection in the genus Alnus and it can be used for preliminary screening in hybrid zones.

Hybridization mediated range expansion and climate change resilience in two keystone tree species of boreal forests.

Current global climate change is expected to affect biodiversity negatively at all scales leading to mass biodiversity loss. Many studies have shown that the distribution of allele frequencies across a species' range is often influenced by specific genetic loci associated with local environmental variables. This association reflects local adaptation and allele changes at those loci could thereby contribute to the evolutionary response to climate change. However, predicting how species will adapt to climate change from this type of data alone remains challenging. In the present study, we combined exome capture sequences and environmental niche reconstruction, to test multiple methods for assessing local adaptation and climate resilience in two widely distributed conifers, Norway spruce and Siberian spruce. Both species are keystone species of the boreal forest and share a vast hybrid zone. We show that local adaptation in conifers can be detected through allele frequency variation, population-level ecological preferences, and historical niche movement. Moreover, we integrated genetic and ecological information into genetic offset predictive models to show that hybridization plays a central role in expanding the niche breadth of the two conifer species and may help both species to cope better with future changing climates. This joint genetic and ecological analysis also identified spruce populations that are at risk under current climate change.

Genomic islands of speciation harbor genes underlying coloration differences in a pair of Neotropical seedeaters.

Incomplete speciation can be leveraged to associate phenotypes with genotypes, thus providing insights into the traits relevant to the reproductive isolation of diverging taxa. We investigate the genetic underpinnings of the phenotypic differences between Sporophila plumbea and S. beltoni. S. beltoni has only recently been described based, most notably, on differences in bill coloration (yellow vs. black in S. plumbea). Both species are indistinguishable through mtDNA or reduced-representation genomic data, and even whole-genome sequencing revealed low genetic differentiation. Demographic reconstructions attribute this genetic homogeneity to gene flow, despite divergence in the order of millions of generations. We found a narrow hybrid zone in southern Brazil where genetically, yet not phenotypically, admixed individuals appear to be prevalent. Despite the overall low genetic differentiation, we identified three narrow peaks along the genome with highly differentiated SNPs. These regions harbor six genes, one of which is involved in pigmentation (EDN3) and is a candidate for controlling bill color. Within the outlier peaks we found signatures of resistance to gene flow, as expected for islands of speciation. Our study shows how genes related to coloration traits are likely involved in generating prezygotic isolation and establishing species boundaries early in speciation.

Hybrid genomics (short review)

Widespread in nature, hybridization between species has a wide range of consequences, and hybrid zones have long been viewed as “windows into evolutionary processes”. The discovery and understanding of the role of hybridization has led to tremendous success in the study of evolution and speciation. Genomic studies make a major contribution to elucidating the role of hybridization in the evolution of modern and extinct taxa. Based on genomic data, the article briefl y reviews modern methods for detecting hybrids, various aspects of the formation and dynamics of hybrid genomes during separate crosses and in the interspecifi c hybridization zones, as well as the genetic and evolutionary consequences of natural hybridization, and highlights current tasks for further research.

The spatial ecology of Mojave rattlesnakes (Crotalus scutulatus), prairie rattlesnakes (Crotalus viridis), and their hybrids in southwestern New Mexico

Abstract Hybridization between species provides unique opportunities to understand evolutionary processes that are linked to reproductive isolation and, ultimately, speciation. However, the extrinsic factors that limit hybridization are poorly understood for most animal systems. Although the spatial ecology of individuals in natural habitats is fundamental to shaping reproductive success and survival, analyses of the spatial ecology of hybrids and their parental groups are rarely reported. Here, we used radiotelemetry to monitor wild rattlesnakes across an interspecific hybrid zone (Crotalus scutulatus and Crotalus viridis) and measured movement parameters and space use (utilization distributions) of individuals to evaluate the hypothesis that hybridization resulted in transgressive or atypical movement patterns. Unexpectedly, of the spatial metrics we investigated, we found that hybrids were very similar to parental individuals. Nonetheless, hybrids did show increased patchiness of core utilization distributions, but this result is likely to be driven by increased habitat patchiness in the hybrid zone. Overall, we did not find evidence for overt extrinsic barriers to hybridization associated with spatial ecology; thus, we suggest that the close evolutionary history between the two parental species and their ecological and behavioural similarities are likely to increase the probability of hybridization events in this unique region of New Mexico.

Do Genetic Loci that Cause Reproductive Isolation in the Lab Inhibit Gene Flow in Nature?

The genetic dissection of reproductive barriers between diverging lineages provides enticing clues into the origin of species. One strategy uses linkage analysis in experimental crosses to identify genomic locations involved in phenotypes that mediate reproductive isolation. A second framework searches for genomic regions that show reduced rates of exchange across natural hybrid zones. It is often assumed that these approaches will point to the same loci, but this assumption is rarely tested. In this perspective, we discuss the factors that determine whether loci connected to postzygotic reproductive barriers in the laboratory are inferred to reduce gene flow in nature. We synthesize data on the genetics of postzygotic isolation in house mice, one of the most intensively studied systems in speciation genetics. In a rare empirical comparison, we measure the correspondence of loci tied to postzygotic barriers via genetic mapping in the laboratory and loci at which gene flow is inhibited across a natural hybrid zone. We find no evidence that the two sets of loci overlap beyond what is expected by chance. In light of these results, we recommend avenues for empirical and theoretical research to resolve the potential incongruence between the two predominant strategies for understanding the genetics of speciation.

Nectary microstructure and nectar production in two species of Cephalocereus (Cactaceae) and their natural hybrid

Variation in the quantity and quality of reward substances produced by flowers influences reproductive success in angiosperms. In this study, a comparative analysis of nectaries and nectar traits (volume, concentration and sugar composition) was performed on the hybrid Cephalocereus columna-trajani × C. tetetzo and its parental species to identify phenotypic variation that could affect reproductive success. The presence of nectarostomata and two types of nectary cells are significant, as well as the presence of xylem and phloem. Furthermore, possible interspecific differences in the amount of starch and mucilage may be relevant. Interspecific variation in nectar volume and concentration was found, with the hybrid exhibiting the highest concentration. All three taxa presented hexose-dominated nectar. In conclusion, C. columna-trajani × C. tetetzo, like its parental species has a functional and anatomically well-formed nectary. Differences in traits such as the amount of starch and mucilage associated with nectar volume, in combination with differences in nectar concentration could have potential consequences for fitness. Our results confirm previous reports on the development of functional structures in hybrid flowers and serve as the basis for future studies testing whether the differences observed here govern the dynamics of competition in hybridization zones.