Reproductive Isolation Research Articles

The only purple-flower species in Actaea L. is pollinated by a hornet

Abstract Examining the pollination biology of plant species is not only crucial for enhancing our understanding of their reproductive biology, but also essential for elucidating their adaptation and evolutionary history. Here, we investigated the breeding system and pollination biology of two closely related species in Actaea. The flower of A. purpurea is unique in the genus with purple and chartaceous (paper-like) sepals, fewer stamens with yellow anthers and purple filaments. Through three seasons of field observation and exclusion experiments, we determined that A. purpurea was primarily pollinated by a hornet species, Vespa bicolor, which also served as the most efficient pollinator. In contrast, A. japonica was primarily pollinated by large flies. Actaea purpurea exhibited a significantly higher cumulative nectar volume than A. japonica, which could be a crucial factor in attracting V. bicolor. A control experiment further demonstrated that removing the nectar leaf (petal) significantly decreased the visiting frequency of V. bicolor. Breeding system studies revealed that both species were self-compatible, yet they primarily underwent outcrossing in natural habitats. Our study presents a compelling case of possible pollinator shift in A. purpurea accompanied by morphological divergence. A more in-depth investigation of this system would offer crucial insights into the extent to which pollinators are involved in the plant speciation process and whether they contribute to reproductive isolation between closely related species.

Genomic evidence reveals three W-autosome fusions in Heliconius butterflies.

Sex chromosomes are evolutionarily labile in many animals and sometimes fuse with autosomes, creating so-called neo-sex chromosomes. Fusions between sex chromosomes and autosomes have been proposed to reduce sexual conflict and to promote adaptation and reproductive isolation among species. Recently, advances in genomics have fuelled the discovery of such fusions across the tree of life. Here, we discovered multiple fusions leading to neo-sex chromosomes in the sapho subclade of the classical adaptive radiation of Heliconius butterflies. Heliconius butterflies generally have 21 chromosomes with very high synteny. However, the five Heliconius species in the sapho subclade show large variation in chromosome number ranging from 21 to 60. We find that the W chromosome is fused with chromosome 4 in all of them. Two sister species pairs show subsequent fusions between the W and chromosomes 9 or 14, respectively. These fusions between autosomes and sex chromosomes make Heliconius butterflies an ideal system for studying the role of neo-sex chromosomes in adaptive radiations and the degeneration of sex chromosomes over time. Our findings emphasize the capability of short-read resequencing to detect genomic signatures of fusion events between sex chromosomes and autosomes even when sex chromosomes are not explicitly assembled.

Ecological niche contributes to the persistence of the western × glaucous-winged gull hybrid zone.

Hybrid zones occur in nature when populations with limited reproductive barriers overlap in space. Many hybrid zones persist over time, and different models have been proposed to explain how selection can maintain hybrid zone stability. More empirical studies are needed to elucidate the role of ecological adaptation in maintaining stable hybrid zones. Here, we investigated the role of exogenous factors in maintaining a hybrid zone between western gulls (Larus occidentalis) and glaucous-winged gulls (L. glaucescens). We used ecological niche models (ENMs) and niche similarity tests to quantify and examine the ecological niches of western gulls, glaucous-winged gulls, and their hybrids. We found evidence of niche divergence between all three groups. Our results support the bounded superiority model, providing further evidence that exogenous selection favoring hybrids may be an important factor in maintaining this stable hybrid zone.

Analysis of the Relationship between Adolescents Need for Reproductive Health Services and Barriers to Access to Reproductive Health Services for Adolescents in Bata Laiworu District, Muna Regency

Introduction: Reproductive health problems are of particular concern, especially those that occur in Indonesian teenagers, both in terms of knowledge and actions for prevention and treatment that need to be taken into account. Based on Indonesian Demographic Health Survey Data, only a small number of young women aged 15 to 19 years have sufficient knowledge. Negative impacts on adolescent reproductive health can be avoided with good sexual and reproductive health information, both directly and indirectly, therefore it is interesting for researchers to conduct an analysis of the relationship between adolescent needs for reproductive health services and barriers to access to adolescent reproductive health services in Bata Laiworu District, Regency. Muna Method: Quantitative observational analytical research using a cross sectional approach, a population of 1,088 and a sample size of 285 students using lemeshow. The sampling method uses Proportional Random Sampling. Result: Based on the Chi Square test that has been carried out, it can be seen (continuity correction) with a p value of 0.000<0.05, meaning there is a relationship between adolescent needs and the use of adolescent reproductive health services. Conclusion: It is necessary for schools to consider including a reproductive health curriculum for students through more in-depth counseling guidance.

Combining biotelemetry and genetics provides complementary insights relevant to the management and conservation of a freshwater predator (Esox lucius) living in brackish lagoons

To inform the management of wild fish populations, it is equally important to understand both the ecological connectivity of habitat patches, apparent at annual and seasonal scales, and the genetic connectivity, emerging at evolutionary scales across generations. Ecological connectivity indicates the potential for rapid recolonization upon local depletion, while genetic connectivity informs about the conservation needs related to the evolution of subpopulations and ecotypes in metapopulations. We combined acoustic biotelemetry and pooled-genome sequencing to study a northern pike (Esox lucius) population as a model of a freshwater piscivore that inhabits a network of shallow brackish lagoons in the southern Baltic Sea. We found limited ecological connectivity among genetically similar subpopulations of pike, suggesting a metapopulation structure characterized by discrete local subpopulations with infrequent migrations between them. Connectivity of different lagoons increased during spawning, suggesting directed spawning migrations to either freshwater rivers or low salinity patches in connected lake-like bays. Spawning site fidelity to either brackish or freshwater spawning sites was observed, further contributing to the reproductive isolation of certain subpopulations. The genetic population structure aligned with salinity gradients and geographical distance and was significant between pairs of rivers draining into the lagoon network, but it was unrelated to ecological connectivity. The results collectively suggest that local subpopulations may not rapidly replenish upon local depletion and that even weak connectivity among subpopulations was sufficient to maintain genetic homogeneity across lagoons with similar salinity levels. Effective management and conservation of species forming metapopulations, such as the coastal northern pike studied here, necessitate localized approaches that adapt fishing mortality to local abundance and promote access to specific habitats, especiallyrivers, during spawning to conserve the entire genetic biodiversity and foster resilience of the metapopulation.

Not so cryptic-differences between mating calls of Hyla arborea and Hyla orientalis from Bulgaria.

Anurans are among the most vocally active vertebrate animals and emit calls with different functions. In order to attract a mate, during the breeding season male frogs produce mating calls which have species-specific structure and parameters, and have been successfully used to resolve issues in taxonomy and phylogenetic relations. This is particularly useful when closely related taxa are concerned, as many species are morphologically almost identical, but still their status is well-supported by molecular and genetic data, suggesting the existence of mechanisms for reproductive isolation. Such is the case for treefrogs from the Hyla arborea group, which are now recognized as several distinct species. The present study aims to establish differences in call parameters between the European tree frog, Hyla arborea, and the Eastern tree frog, Hyla orientalis, which both occur on the territory of Bulgaria. Using autonomous audio loggers, calls from six sites (three in the range of H. arborea and three in the range of H. orientalis) were recorded between 7 p.m. and 12 a.m. during the breeding season in 2020-2023. The following parameters in a total of 390 mating calls were analyzed: call count, pulse count, call series duration, call period, peak (dominant) frequency, entropy. Results indicated that sites formed two distinct groups, which corresponded to the known distribution ranges of H. arborea and H. orientalis. The first two components of the PCA explained 71% of the total variance, with variables call count, call series duration, peak frequency and entropy being most important for differentiation between the sites. This study presents the first attempt to differentiate between the calls of these two sister taxa, which both fall within the "short-call treefrogs" group, and results are discussed in terms of known data for mating calls in Hyla sp., as well as limitations and future perspectives.

The neurodevelopmental genes alan shepard and Neuroglian contribute to female mate preference in African Drosophila melanogaster.

Mate choice is a key trait that determines fitness for most sexually reproducing organisms, with females often being the choosy sex. Female preference often results in strong selection on male traits that can drive rapid divergence of traits and preferences between lineages, leading to reproductive isolation. Despite this fundamental property of female mate choice, very few loci have been identified that contribute to mate choice and reproductive isolation. We used a combination of population genetics, quantitative complementation tests, and behavioural assays to demonstrate that alan shepard and Neuroglian contribute to female mate choice, and could contribute to partial reproductive isolation between populations of Drosophila melanogaster. Our study is among the first to identify genes that contribute to female mate preference in this historically important system, where female preference is an active premating barrier to reproduction. The identification of loci that are primarily known for their roles in neurodevelopment provides intriguing questions of how female mate preference evolves in populations via changes in sensory system and higher learning brain centres.

Assessing the temporal isolation barrier between two closely related species of the genus Fissurella off the southwestern coast of South America

ABSTRACT Knowledge of the reproductive isolation mechanisms that prevent gene flow between species is essential for understanding the process of speciation. In this study, we investigate the spawning periods in Fissurella cumingi and F. latimarginata, two closely related sympatric species of keyhole limpets with external fertilization, as a potential prezygotic barrier between them. The reproductive cycle was assessed in two distinct locations in northern Chile using the gonadosomatic index (GSI) and gonadal histology. Species, locality, and sex were designated as predictive variables, while average GSI values, as well as those of partial and total spawning, were considered dependent variables. Both species exhibit a continuous gametogenic cycle throughout the year, characterized by multiple spawning events. A generalized linear model (GLM) factorial ANOVA design revealed that neither species, locality nor sex were significant factors with respect to the GSI and partial spawning scores. However, sex was found to be the only significant factor related to total spawning. No significant interactions were observed among predictive factors. These findings suggest that reproductive periods may not serve as effective barriers to gamete encounters between species. Our study prompts questions about how closely-related externally-fertilizing species maintain reproductive isolation.

Microscopic observation and transcriptome analysis provide insights into mechanisms of hybrid incompatibility in Rhododendron

Hybridization across species has been widely utilized to improve agriculturally important traits in Rhododendron. These interspecific hybridizations are often inhibited by reproductive barriers at the prezygotic and postzygotic stages. Thus, in order to character the barriers and reveal the underlying regulation mechanism of interspecific hybrid incompatibility in Rhododendron, we compared the pollen germination and pollen tube growth, fruit growth, seed number and germination rate between intraspecific (Rhododendron ‘XXL’ × R. ‘XXL’) and interspecific (R. ‘XXL’ × R. delavayi) hybridizations. After pollination, the pollen germination rate and pollen tube growth rate of interspecific hybridization (Inter) were always higher than that of intraspecific hybridization (Intra) during the whole observation period. Conversely, the fruits and ovules of Intra grew faster than that of Inter after 20 DAP. The ratio of shrunken embryos of Inter was 73.27 ± 10.19 %, which was significantly higher than that of Intra with 28.41 ± 8.33 % at 40 DAP. Moreover, the fruit setting rate, the average number of seeds per fruit and seed germination rate of Intra were significantly higher than that of Inter at 150 DAP. These results indicated that the barriers of interspecific incompatibility between R. ‘XXL’ × R. delavayi could be mainly ascribed to post-fertilization isolation. Consequently, RNA-Seq analysis on ovaries at three developmental stages from two hybridizations indicated that plant hormone signal transduction, flavonoid biosynthesis, MAPK signaling pathway and plant-pathogen interaction pathways might play important roles in regulation of hybrid incompatibility between R. ‘XXL’ × R. delavayi. The results can provide a basis for the study of the hybrid incompatibility mechanism of Rhododendron, and provide useful information for the breeding and conservation of genetic resources of Rhododendron.

Introgression of the gamete recognition molecule, bindin, in the sea urchin Diadema.

Hybridization is important in evolution, because it is a necessary (though not sufficient) step in the introgression of potentially adaptive variation between species. Bindin is a gamete recognition protein in echinoids and asteroids, capable of blocking cross-fertilization between species to varying degrees. Four species of the sea urchin genus Diadema are broadly sympatric in the Indo-Pacific: D. paucispinum, D. savignyi, D. clarki, and D. setosum. Data from three published studies, one of identification of hybrids through allozymes, one of the phylogeography of mitochondrial DNA, and one of the phylogeny of bindin, were combined to assess the degree of bindin introgression between these four species. I analyzed sequences of the ATPase 8 and ATPase 6 mitochondrial genes and of bindin, sampled throughout the species ranges, with an Isolation-Migration algorithm, IMa3. IMa3 uses a coalescent approach to produce Bayesian estimates of effective population sizes and gene flow between populations. The results showed that bindin alleles coalesce completely within the species bounds of D. clarki, and of D. setosum. The sister species D. paucispinum and D. savignyi, however, were estimated as having exchanged a bindin allele at an average of every one to two-and-a-half generations since they speciated from each other. As the allozyme study detected nine hybrids between three of these species in Okinawa (most of them between D. setosum and D. savignyi) in a single sample, hybrids between these species are produced, but bindin does not introgress. Therefore, bindin must not be efficient in blocking heterospecific fertilizations. Complete, or almost complete, reproductive isolation between species of Diadema must result from low hybrid fitness.

Ploidy as a leaky reproductive barrier: mechanisms, rates and evolutionary significance of interploidy gene flow.

Whole genome duplication (polyploidization) is a dominant force in sympatric speciation, particularly in plants. Genome doubling instantly poses a barrier to gene flow owing to the strong crossing incompatibilities between individuals differing in ploidy. The strength of the barrier, however, varies from species to species and recent genetic investigations revealed cases of rampant interploidy introgression in multiple ploidy-variable species. Here, we review novel insights into the frequency of interploidy gene flow in natural systems and summarize the underlying mechanisms promoting interploidy gene flow. Field surveys, occasionally complemented by crossing experiments, suggest frequent opportunities for interploidy gene flow, particularly in the direction from diploid to tetraploid, and between (higher) polyploids. However, a scarcity of accompanying population genetic evidence and a virtual lack of integration of these approaches leave the underlying mechanisms and levels of realized interploidy gene flow in nature largely unknown. Finally, we discuss potential consequences of interploidy genome permeability on polyploid speciation and adaptation and highlight novel avenues that have just recently been opened by the very first genomic studies of ploidy-variable species. Standing in stark contrast with rapidly accumulating evidence for evolutionary importance of homoploid introgression, similar cases in ploidy-variable systems are yet to be documented. The genomics era provides novel opportunity to re-evaluate the role of interploidy introgression in speciation and adaptation. To achieve this goal, interdisciplinary studies bordering ecology and population genetics and genomics are needed.

Maintenance of a narrow hybrid zone between native and introduced red foxes (Vulpes vulpes) despite conspecificity and high dispersal capabilities.

Human-facilitated introductions of nonnative populations can lead to secondary contact between allopatric lineages, resulting in lineage homogenisation or the formation of stable hybrid zones maintained by reproductive barriers. We investigated patterns of gene flow between the native Sacramento Valley red fox (Vulpes vulpes patwin) and introduced conspecifics of captive-bred origin in California's Central Valley. Considering their recent divergence (20-70 kya), we hypothesised that any observed barriers to gene flow were primarily driven by pre-zygotic (e.g. behavioural differences) rather than post-zygotic (e.g. reduced hybrid fitness) barriers. We also explored whether nonnative genes could confer higher fitness in the human-dominated landscape resulting in selective introgression into the native population. Genetic analysis of red foxes (n = 682) at both mitochondrial (cytochrome b + D-loop) and nuclear (19,051 SNPs) loci revealed narrower cline widths than expected under a simulated model of unrestricted gene flow, consistent with the existence of reproductive barriers. We identified several loci with reduced introgression that were previously linked to behavioural divergence in captive-bred and domestic canids, supporting pre-zygotic, yet possibly hereditary, barriers as a mechanism driving the narrowness and stability of the hybrid zone. Several loci with elevated gene flow from the nonnative into the native population were linked to genes associated with domestication and adaptation to human-dominated landscapes. This study contributes to our understanding of hybridisation dynamics in vertebrates, particularly in the context of species introductions and landscape changes, underscoring the importance of considering how multiple mechanisms may be maintaining lineages at the species and subspecies level.

Can developmental plasticity shape sexual competition and promote reproductive isolation?

Abstract Environmental factors, such as dietary nutrients, can shape the expression of developmentally plastic sexual traits in many species. However, while there has been extensive research into the developmental plasticity of sexual traits at the individual level, the broader consequences of this variation at the population scale remain poorly understood. Here, we asked whether plastic responses to the developmental environment can shape sexual competition and initiate reproductive isolation between populations. We reared neriid flies, Telostylinus angusticollis, on nutrient-rich and nutrient-poor larval diets, generating adult flies that differed in body size and secondary sexual trait expression. We then investigated sexual competition in experimental populations from each developmental environment and tested for reproductive isolation between flies from mismatched environments. We found that, compared with poor-diet populations, rich-diet populations exhibited more frequent and escalated male–male combat and more frequent mating and mate-guarding. However, we found no evidence that sexual selection was affected by the developmental environment. Mismatched female–male pairs tended to take longer to mate and rich-diet females often rejected poor-diet males, but mismatched pairs were not less likely to mate within 1 h or produce viable offspring. Our findings suggest that developmental plasticity could generate dramatic differences in sexual competition between populations and could contribute to reproductive isolation.

Bulked Segregant RNA-Seq Analysis of Pollinated Pistils Reveals Genes Influencing Spikelet Fertility in Rice

Prezygotic isolation is important for successful fertilization in rice, significantly affecting yield. This study focused on F5:6 generation plants derived from inter-subspecific crosses (Nipponbare × KDML105) with low (LS) and high seed-setting rates (HS), in which normal pollen fertility was observed. However, LS plants showed a reduced number of pollen grains adhering to the stigma and fewer pollen tubes reaching the ovules at 4‒5 h post-pollination, compared with HS plants. Bulked segregant RNA-Seq analysis of pollinated pistils from the HS and LS groups revealed 249 and 473 differentially expressed genes (DEGs), respectively. Kyoto Encyclopedia of Genes and Genomes analysis of the HS and LS- specific DEGs indicated enrichment in metabolic pathways, pentose and glucuronate interconversions, and flavonoid biosynthesis. Several of these DEGs exhibited co-expression with pollen development genes and formed extensive clusters of co-expression networks. Compared with LS pistils, enzyme genes controlling pectin degradation, such as OsPME35 and OsPLL9, showed similar expression patterns, with higher levels in HS pistils pre-pollination. Os02g0467600, similar to cinnamate 4-hydroxylase gene (CYP73), involved in flavonoid biosynthesis, displayed higher expression in HS pistils post-pollination. Our findings suggest that OsPME35, OsPLL9, and Os02g0467600 contribute to prezygotic isolation by potentially modifying the stigma cell wall (OsPME35 and OsPLL9) and controlling later processes such as pollen-stigma adhesion (Os02g0467600) genes. Furthermore, several DEGs specific to HS and LS were co-localized with QTLs and functional genes associated with spikelet fertility. These findings provide valuable insights for further research on rice spikelet fertility, ultimately contributing to the development of high-yielding rice varieties.

Lake Baikal amphipods and their genomes, great and small.

Endemic amphipods (Crustacea: Amphipoda) of Lake Baikal represent an outstanding example of large species flocks occupying a wide range of ecological niches and originating from a handful of ancestor species. Their development took place at a restricted territory and is thus open for comprehensive research. Such examples provide unique opportunities for studying behavioral, anatomic, or physiological adaptations in multiple combinations of environmental conditions and thus attract considerable attention. The existing taxonomies of this group list over 350 species and subspecies, which, according to the molecular phylogenetic studies of marker genes, full transcriptomes and mitochondrial genomes, originated from at least two introductions into the lake. The studies of allozymes and marker genes have revealed a significant cryptic diversity in Baikal amphipods, as well as a large variance in genetic diversity within some morphological species. Crossing experiments conducted so far for two morphological species suggest that the differences in the mitochondrial marker (cytochrome c oxidase subunit I gene) can potentially be applied for making predictions about reproductive isolation. For about one-tenth of the Baikal amphipod species, nuclear genome sizes and chromosome numbers are known. While genome sizes vary within one order of magnitude, the karyotypes are relatively stable (2n = 52 for most species studied). Moreover, analysis of the diversity of repeated sequences in nuclear genomes showed significant between-species differences. Studies of mitochondrial genomes revealed some unusual features, such as variation in length and gene order, as well as duplications of tRNA genes, some of which also underwent remolding (change in anticodon specificity due to point mutations). The next important steps should be (i) the assembly of whole genomes for different species of Baikal amphipods, which is at the moment hampered by complicated genome structures with high repeat content, and (ii) updating species taxonomy taking into account all the data.

Variations in the phenological patterns of a caddisfly inhabiting the same mountain massifs: Life‐history differences in different altitudinal zones

AbstractOrganisms inhabiting mountainous regions can experience large vertical environmental changes, and show different ecological characteristics between altitudes, thus facilitating allopatric fragmentation even in geographically close populations. This study compared the life‐history patterns of a species of limnephilid caddisfly, Asynarchus sachalinensis, in several genetically differentiated populations between alpine and sub‐alpine zones in a temperate mountainous region. We showed that in the sub‐alpine populations, larval development started earlier with increasing water temperature in spring, and adult emergence was also earlier. The occurrence of adults was extremely low in mid‐summer, probably due to summer diapause, followed by a larger number of ovary‐developed females in autumn. On the other hand, in the alpine zone, increasing water temperature was delayed compared to the sub‐alpine zone, and larval development occurred from early to mid‐summer. Adult emergence and ovary‐developed individuals were concentrated in mid‐summer. Hence, summer diapause was not observed. These results indicated life‐history differences between genetically differentiated populations at different altitudes. As the timing of adult occurrence and ovarian developmental patterns differ between populations at different altitudes, it is possible that reproductive isolation is facilitated or maintained between populations.

Selection on the joint actions of pairs leads to divergent adaptation and coadaptation of care-giving parents during pre-hatching care.

The joint actions of animals in partnerships or social groups evolve under both natural selection from the wider environment and social selection imposed by other members of the pair or group. We used experimental evolution to investigate how jointly expressed actions evolve upon exposure to a new environmental challenge. Our work focused on the evolution of carrion nest preparation by pairs of burying beetles Nicrophorus vespilloides, a joint activity undertaken by the pair but typically led by the male. In previous work, we found that carrion nest preparation evolved to be faster in experimental populations without post-hatching care (No Care: NC lines) than with post-hatching care (Full Care: FC lines). Here, we investigate how this joint activity evolved. After 15 generations of experimental evolution, we created heterotypic pairs (NC females with FC males and NC males with FC females) and compared their carrion nest making with homotypic NC and FC pairs. We found that pairs with NC males prepared the nest more rapidly than pairs with FC males, regardless of the female's line of origin. We discuss how social coadaptations within pairs or groups could act as a post-mating barrier to gene flow.

Maternal investment evolves with larger body size and higher diversification rate in sharks and rays

Across vertebrates, live bearing evolved at least 150 times from ancestral egg laying into diverse forms and degrees of prepartum maternal investment.1,2 A key question is how reproductive diversity arose and whether reproductive diversification underlies species diversification.3,4,5,6,7,8,9,10,11 To test this, we evaluate the most basal jawed vertebrates: the sharks, rays, and chimaeras, which have one of the greatest ranges of reproductive and ecological diversity among vertebrates.2,12 We reconstruct the sequence of reproductive mode evolution across a phylogeny of 610 chondrichthyans.13 We reveal egg laying as ancestral, with live bearing evolving at least seven times. Matrotrophy evolved at least 15 times, with evidence of one reversal. In sharks, transitions to live bearing and matrotrophy are more prevalent in larger-bodied tropical species. Further, the evolution of live bearing is associated with a near doubling of the diversification rate, but there is only a small increase associated with the appearance of matrotrophy. Although pre-copulatory sexual selection is associated with increased rates of speciation in teleosts,3 sexual size dimorphism in chondrichthyans does not appear to be related to sexual selection,14,15 and instead we find increased rates of speciation associated with the colonization of novel habitats. This highlights a potential key difference between chondrichthyans and other fishes, specifically a slower rate of evolution of reproductive isolation following speciation, suggesting different rate-limiting mechanisms for diversification between these clades.16 The chondrichthyan diversification and radiation, particularly throughout shallow tropical shelf seas and oceanic pelagic habitats, appear to be associated with the evolution of live bearing and proliferation of a wide range of maternal investment in developing offspring.

Song and genetic divergence within a subspecies of white-crowned sparrow (Zonotrichia leucophrys nuttalli).

Animal culture evolves alongside genomes, and the two modes of inheritance-culture and genes-interact in myriad ways. For example, stable geographic variation in culture can act as a reproductive barrier, thereby facilitating genetic divergence between "cultural populations." White-crowned sparrows (Zonotrichia leucophrys) are a well-established model species for bird song learning and cultural evolution, as they have distinct, geographically discrete, and culturally transmitted song types (i.e., song dialects). In this study, we tested the hypothesis that divergence between culturally transmitted songs drives genetic divergence within Nuttall's white-crowned sparrows (Z. l. nuttalli). In accordance with sexual selection theory, we hypothesized that cultural divergence between mating signals both preceded and generated genetic divergence. We characterized the population structure and song variation in the subspecies and found two genetically differentiated populations whose boundary coincides with a major song boundary at Monterey Bay, California. We then conducted a song playback experiment that demonstrated males discriminate between songs based on their degree of divergence from their local dialect. These results support the idea that discrimination against non-local songs is driving genetic divergence between the northern and southern populations. Altogether, this study provides evidence that culturally transmitted bird songs can act as the foundation for speciation by sexual selection.

Reproductive isolation arises during laboratory adaptation to a novel hot environment

BackgroundReproductive isolation can result from adaptive processes (e.g., ecological speciation and mutation-order speciation) or stochastic processes such as “system drift” model. Ecological speciation predicts barriers to gene flow between populations from different environments, but not among replicate populations from the same environment. In contrast, reproductive isolation among populations independently adapted to the same/similar environment can arise from both mutation-order speciation or system drift.ResultsIn experimentally evolved populations adapting to a hot environment for over 100 generations, we find evidence for pre- and postmating reproductive isolation. On one hand, an altered lipid metabolism and cuticular hydrocarbon composition pointed to possible premating barriers between the ancestral and replicate evolved populations. On the other hand, the pronounced gene expression differences in male reproductive genes may underlie the postmating isolation among replicate evolved populations adapting to the same environment with the same standing genetic variation.ConclusionOur study confirms that replicated evolution experiments provide valuable insights into the mechanisms of speciation. The rapid emergence of the premating reproductive isolation during temperature adaptation showcases incipient ecological speciation. The potential evidence of postmating reproductive isolation among replicates gave rise to two hypotheses: (1) mutation-order speciation through a common selection on early fecundity leading to an inherent inter-locus sexual conflict; (2) system drift with genetic drift along the neutral ridges.