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Evidence for historical introgression along a contact zone between two species of char (Pisces: Salmonidae) in northwestern North America.

Phylogeographic analyses can yield valuable insights into the geographic and historical contexts of contact and hybridization between taxa. Two species of char (Salmonidae), Dolly Varden (Salvelinus malma) and bull trout (S. confluentus) have largely parapatric distributions in watersheds of northwestern North America. They are, however, sympatric in several localities and hybridization and some introgression occurs across a broad area of contact. We conducted a comparative phylogenetic analysis of Dolly Varden and bull trout to gain a historical perspective of hybridization between these species and to test for footprints of historical introgression. We resolved two major Dolly Varden mitochondrial DNA (mtDNA) clades (with 1.4-2.2% sequence divergence between haplotypes) that had different geographical distributions. Clade N is distributed across most of the range of Dolly Varden, from southern British Columbia through to the Kuril Islands in Asia. Clade S had a much more limited distribution, from Washington state, at the southern limit of the Dolly Varden range, to the middle of Vancouver Island. The distribution and inferred ages of the mtDNA clades suggested that Dolly Varden survived the Wisconsinan glaciation in a previously unsuspected refuge south of the ice sheet, and that Dolly Varden and bull trout were probably in continuous contact over most of the last 100,000 years. When bull trout were included in the phylogenetic analysis, however, the mtDNA of neither species was monophyletic: Clade S Dolly Varden clustered within the bull trout mtDNA clade. This pattern was discordant with two nuclear phylogenies produced (growth hormone 2 and rRNA internal transcribed sequence 1), in which Dolly Varden and bull trout were reciprocally monophyletic. This discordance between mtDNA- and nDNA-based phylogenies indicates that historical introgression of bull trout mtDNA into Dolly Varden occurred. Percent sequence divergence within these introgressed Dolly Varden (clade S) was 0.2-0.6%, implying that the introgression occurred prior to the most recent glaciation. Our analysis and other evidence of contact between divergent lineages in northwestern North America strongly suggests that the area may be the site of previously unsuspected suture zones of aquatic biotas.

Animals↗

[Analysis of inheritance of morphological and biochemical characters introgressed into common wheat from Aegilops speltoides Tausch].

Genetic control of some morphological traits and the gliadin composition were examined in plants of two lines of common wheat carrying genes introgressed from the wild diploid cereal Aegilops speltoides. Leaf hairiness was shown to be controlled by a single introgressed dominant gene that was not allelic to the known common wheat gene Hl1. Waxlessness of the whole plant is controlled by the introgressed from Ae. speltoides inhibitor gene allelic to gene W1 located on chromosome 2B. This gene was epistatic to the introgressed gene controlling spike waxlessness. The introgressed gene of spike color was shown to be allelic to Rg1 located on chromosome 1B of common wheat. However, the former gene proved to be linked to an allele of the Gli-B1 locus other than in wheat.

Alleles↗

Cryptic interspecific introgression and genetic differentiation within Gossypium aridum (Malvaceae) and its relatives.

Interspecific gene flow is increasingly recognized as an important evolutionary phenomenon in plants. A surprising observation is that historical introgression is often inferred between species that presently have geographic and reproductive barriers that would appear to prohibit the inferred sexual exchange. A striking example concerns Gossypium aridum (subsection Erioxylum); previous analyses have shown that populations from Colima (southwestern Mexico) have a chloroplast genome (cpDNA) similar to that of a different taxonomic subsection (Integrifolia) that presently is confined to Baja California and the Galapagos Islands, whereas other G. aridum populations share a cpDNA lineage with each other and with other species in subsection Erioxylum. To evaluate further the possibility that this cpDNA evidence reflects introgression as opposed to some other evolutionary process, as well as to explore patterns of genetic diversity and similarity in both subsections, we conducted amplified fragment-length polymorphism (AFLP) analysis using 50 populations representing all seven species in the two subsections. Genetic diversity is high in G. aridum, and is strongly correlated with geography, as are similarities among the five species in subsection Erioxylum. This subsection is genetically distant from the two species in subsection Integrifolia, whose populations are highly similar inter se. Populations of G. aridum from Colima are genetically distinct from the remainder of the species, and exhibit a comparatively high frequency of AFLP fragments that otherwise are diagnostic of the Integrifolia lineage. These data implicate intersubsectional introgression between presently allopatric and genetically isolated clades, giving rise to a morphologically cryptic, introgressant entity. Biogeographic considerations suggest that this history was initiated following migration of one or more seeds from Baja California to the Colima coast, perhaps during the Pleistocene. We suggest that cryptic and seemingly improbable interspecific introgression and molecular differentiation may be more common than appreciated in angiosperm evolution.

Analysis of Variance↗

Genomic Footprints of Historical Introgression Between Ancient Lineages of Wild Oryza AA-Genome Species With Widely Separated Contemporary Distributions.

Phylogenetic incongruence is increasingly recognized as pervasive, yet the extent to which reticulate evolution occurs between groups separated by substantial geographical distances and deep phylogenetic divergence remains poorly characterized. In the Oryza AA-genome group-a model for plant speciation and domestication-the traditional bifurcation model posits that Australian Oryza meridionalis and African Oryza longistaminata occupy basal branches, distinct from the more recently diversified monophyletic clade comprising Asian and other African lineages, including major cultivars. However, recent evidence from endogenous viral sequences has hinted at unexpected genetic relatedness between African O. longistaminata and Asian Oryza sativa, which are geographically and phylogenetically distant. Here, we conducted a genome-wide survey across 11 Oryza species to systematically identify genomic regions exhibiting phylogenetic incongruence. Widespread phylogenetic discordance was observed, notably involving genomic segments in which O. longistaminata showed phylogenetic proximity to Asian species, contradicting their established deep divergence. To distinguish between introgression and incomplete lineage sorting, we performed four-taxon ABBA-BABA tests, which provided statistical support for introgression. Furthermore, divergence time estimates for these incongruent regions were younger than the species divergence times, suggesting historical introgression between the ancestors of lineages that are currently separated by vast geographical distances. Systematic assessments indicated that potential analytical artifacts, such as compositional bias and substitution saturation, were unlikely to explain the observations. These convergent lines of evidence suggest that ancient introgression had occurred between currently geographically separated and evolutionarily divergent Oryza lineages, leaving detectable footprints across their modern genomes.

Oryza↗

Strategies to optimize marker-assisted introgression of multiple unlinked QTL.

To optimize designs to implement marker-assisted introgression programs aiming to introgress three unlinked quantitative trait loci (QTL), the present paper studies different alternatives versus a traditional backcross or intercross phase. Four alternative backcross strategies appear to be more advantageous by having 50% less genotyping load than a traditional backcross strategy tracking all three QTL at a time through a single line. A multiplication phase following the selection of homozygous animals at the three QTL as an intercross alternative allows doubling of the number of homozygous animals in a mouse model compared with the first intercross generation. Within the same model, a second intercross alternative with individuals carrying all three QTL at the first intercross results in a 12-fold increase in the number of homozygous animals obtained in the first intercross generation. The same ranges of decrease are observed in the number of animals to be genotyped and the number of genotypings when targeting a fixed number of homozygous animals. An option, with two lines each carrying two QTL through the backcross phase and coupled with the second intercross alternative, appears to be the best introgression alternative. This option requires 76% fewer genotypings, 68% fewer animals to be genotyped, and costs 75% less than an option in which all three QTL are introgressed through a single line.

Animals↗

Genetics of hybrid inviability and sterility in Drosophila: dissection of introgression of D. simulans genes in D. melanogaster genome.

Interspecific crosses between Drosophila melanogaster and Drosophila simulans usually produce sterile unisexual hybrids. The barrier preventing genetic analysis of hybrid inviability and sterility has been taken away by the discovery of a D. simulans strain which produces fertile female hybrids. D. simulans genes in the cytological locations of 21A1 to 22C1-23B1 and 30F3-31C5 to 36A2-7 have been introgressed into the D. melanogaster genetic background by consecutive backcrosses. Flies heterozygous for the introgression are fertile, while homozygotes are sterile both in females and males. The genes responsible for the sterility have been mapped in the introgression. The male sterility is caused by the synergistic effect of multiple genes, while the female sterility genes have been localized to a 170 kb region (32D2 to 32E4) containing 20 open reading frames. Thus, the female sterility might be attributed to a single gene with a large effect. We have also found that the Lethal hybrid rescue mutation which prevents the inviability of male hybrids from the cross of D. melanogaster females and D. simulans males cannot rescue those carrying the introgression, suggesting that D. simulans genes maybe non-functional in this hybrid genotype. The genes responsible for the inviability have not been separated from the female sterility genes by recombination.

Animals↗

Genomic in situ hybridization analysis for identification of introgressed segments in alloplasmic lines from Zea mays x Zea diploperennis.

In this study, the tested four alloplasmic inbred lines, a2-4, a2-5, b1-1 and b2-1 were propagated from the same disease resistant individual in the parthenogenetic progenies of Zea mays L. cv. Lu 9 x Zea diploperennis (DP). All the lines except a2-5 were resistant to Helminthosporium turcium Pass and H. maydis Nisik. Introgressed DP segments in these lines were detected by both Southern hybridization and genomic in situ hybridization (GISH). The results of Southern hybridization showed that DP species-specific DNA sequences had been introgressed into the genomes of alloplasmic lines. The Southern hybridization band patterns in all of the tested lines were consistent with those of DP. Genomic in situ hybridization (GISH) signals were detected on 7 different chromosome pairs in lines a2-4 and a2-5, on 5 chromosome pairs in b1-1 and on 4 chromosome pairs in b2-1. The features of introgression, and disease resistant genes in the introgressed segments, as well as the gene silence or elimination in some alloplasmic lines are discussed.

Blotting, Southern↗

Gene introgression into Coffea arabica by way of triploid hybrids (C. arabica x C. canephora).

Interspecific triploid hybrid plants between the tetraploid species Coffea arabica L. and the diploid species C. canephora P. were backcrossed to C. arabica. Although characterised by a low production and an important fruit dropping, all attempted crosses (ie, 6) generated BC(1) progenies. Flow cytometric analysis of the nuclear DNA content revealed that most of the BC1 individuals were nearly tetraploid. Among the male gametes produced by the interspecific triploid hybrids, those presenting a high number of chromosomes appeared strongly favoured. Only pollen mother cells having nearly 22 chromosomes were effective, the others leading to deficient endosperm and fruit dropping. Molecular markers (ie, microsatellite and AFLP) combined with evaluations of morphological characteristics and resistance to leaf rust were applied to verify the occurrence of gene transfer from C. canephora into C. arabica, and to estimate the amount of introgression present in BC(1) individuals. The results reveal a strong deficiency in the C. canephroa alleles indicating a severe counter-selection against the introgression of genetic material from C. canephora into C. arabica by way of triploid hybrids. However, introgressants displaying desirable traits such as a high resistance to leaf rust were obtained. The low level of introgression could be an advantage by facilitating the recovery of the recurrent parent and possibly reducing the number of required backcrosses. On the other hand, this could be a limitation when attempting the transfer of a complex trait or several simply inherited traits.

Coffea↗

Genetic introgression between wild and stocked salmonids and the prospects for using molecular markers in population rehabilitation: the case of the Adriatic grayling (Thymallus thymallus L. 1785).

In the north Adriatic basin, a morphologically and genetically distinct lineage of grayling is found, designated as the Adriatic grayling. In Slovenia, the Adriatic grayling is restricted to the Soca river system, where it is critically endangered. The most pertinent threat is stocking with non-native, highly divergent Sava (Danubian) drainage stock, and this activity has been going on for more than four decades. The present study was designed to characterise the genetic structure of the Adriatic grayling in Slovenia, with particular emphasis on estimating the degree of introgression with non-indigenous stocked grayling. We analysed polymorphism at 154 microsatellite loci in samples representing grayling from the Adriatic and Danubian drainage stock. A relatively high number (12) of alleles, diagnostic for the Adriatic grayling, were identified. However, a correspondence analysis based on individual multilocus genotypes also revealed that there is no distinctive Adriatic group but rather a dispersed multitude of individuals that cannot be unambiguously distinguished from the more homogenous Danubian population. A Bayesian analysis of individual admixture coefficients confirmed this pattern and revealed extensive introgression between the Adriatic grayling and stocked grayling of Danubian origin. Average individual admixture coefficients showed that only between 50 and 60% of the original gene pools remained, and only few non-introgressed indigenous individuals could be identified. Microsatellite-based individual admixture analysis appear to be an important tool for identifying remaining non-introgressed indigenous individuals that could be used for restoring the original populations.

Alleles↗

Differential introgression of uniparentally inherited markers in bison populations with hybrid ancestries.

Historical hybridization between Bison bison (bison) and Bos taurus (cattle) has been well documented and resulted in cattle mitochondrial DNA (mtDNA) introgression, previously identified in six different bison populations. In order to examine Y chromosome introgression, a microsatellite marker (BYM-1) with non-overlapping allele size distributions in bison and cattle was isolated from a bacterial artificial chromosome (BAC) clone, and was physically assigned to the Y chromosome by fluorescence in situ hybridization. BYM-1 genotypes for a sample of 143 male bison from 10 populations, including all six populations where cattle mtDNA haplotypes were previously identified, indicated that cattle Y chromosome introgression had not occurred in these bison populations. The differential permeability of uniparentally inherited markers to introgression is consistent with observations of sterility among first generation hybrid males and a sexual asymmetry in the direction of hybridization favouring matings between male bison and female cattle.

Animals↗

Introgressive hybridization in Rorippa (Brassicaceae): gene flow and its consequences in natural and anthropogenic habitats.

Introgressive hybridization between three Rorippa species (R. amphibia, R. palustris and R. sylvestris) in northern Germany has been studied using isozymes and noncoding chloroplast DNA (trnL/F spacer). Our results provide substantial evidence for different patterns of gene flow in natural and in anthropogenic environments. Hybridization and bi-directional introgression (chloroplast DNA and allozymes) between R. amphibia and R. sylvestris were detected at the river Elbe, which is one of the last rivers in Central Europe showing a natural dynamic of erosion and sedimentation. The natural dynamic of the Elbe leads to periodic habitat disturbance and the temporal breakdown of ecological isolation barriers between R. amphibia and R. sylvestris. However, the high dynamic does not provide the opportunity for persistence of the morphologically intermediate hybrid R. x anceps (R. amphibia x R. sylvestris). We did not find hybrid zones between R. amphibia and R. sylvestris in the more anthropogenic landscape of northwest Germany. However, contact zones between R. amphibia and R. palustris were detected in drainage ditches in northwest Germany. We found substantial evidence for unidirectional introgression of R. palustris markers (chloroplast DNA and allozymes) into R. amphibia in the man-made habitats. The R. amphibia introgressants in the drainage ditches often showed strongly serrate upper cauline leaves instead of the entire upper cauline leaves typical for R. amphibia. We argue that landscape melioration in northwest Germany, particularly the creation of drainage ditches, favoured both hybrid-zone formation and ecotypic differentiation within R. amphibia.

Brassicaceae↗

Adaptive introgression of herbivore resistance traits in the weedy sunflower Helianthus annuus.

The role of hybridization in adaptive evolution is contentious. While many cases of adaptive trait introgression have been proposed, the relevant traits have rarely been identified, resulting in a lack of clear examples of this process. Here, we examine a purported case of adaptive introgression in which the annual sunflower Helianthus annuus annuus has captured alleles from a congener (Helianthus debilis) to form a stabilized hybrid, Helianthus annuus texanus. We tested the hypotheses that herbivore resistance traits have introgressed from H. debilis to H. annuus and have increased adaptation in the latter. In two common gardens, fitness (estimated by seed production) was on average 55% higher in H. a. texanus than in H. a. annuus. For H. a. texanus, three damage traits (of seven tested) differed significantly from the H. a. annuus parent in one or both sites and were shifted in the direction of the more resistant H. debilis. Natural selection favored H. a. annuusxH. debilis BC(1) hybrids (synthesized to mimic the ancestors of H. a. texanus) with H. debilis-like resistance to seed midges Neolasioptera helianthis and to receptacle/seed feeding Lepidoptera at one or both sites. Assuming similar herbivore pressures in the past, these results suggest that introgression of biotic resistance traits was important in the adaptation of H. annuus to central and southern Texas.

Adaptation, Biological↗

KCFtools: rapid alignment-free method for introgression screening and GWAS using k-mer profiles.

MOTIVATION: In the era of multiple genome references, researchers often align sequencing reads against distinct assemblies or even multiple references simultaneously. This enables applications such as the detection of introgressed segments or highly variable genomic regions, which are especially prevalent in large-genome crop species such as lettuce or wheat. However, these applications come at the cost of increased computational burden, inconsistencies in mapping methods, and reduced reproducibility across studies. To address these limitations, we developed KCFtools, a Java-based toolkit that identifies the presence and absence of k-mers in nonoverlapping genomic or transcriptomic windows by comparing query and reference genomes. This alignment-free approach enables the efficient computation of an identity score for each window, thereby facilitating robust detection of introgressed or variable regions across genomes. RESULTS: We systematically evaluated the performance and accuracy of the k-mer-based method implemented in KCFtools, benchmarking it against conventional single nucleotide variation-based introgression detection pipelines. Our results demonstrate that KCFtools effectively captures introgressed segments and structurally diverse regions, even in species with fragmented or highly divergent reference genomes. In addition, we extended KCFtools to generate genotype matrices from k-mer variation tables. These matrices are compatible with genome-wide association studies software and allow the identification of loci associated with phenotypic traits. We showcase the utility of this approach by detecting known and novel associations for downy mildew resistance in lettuce, underscoring the pipeline's potential for high-resolution, reference-agnostic population genetic analysis. AVAILABILITY AND IMPLEMENTATION: https://github.com/sivasubramanics/kcftools.

Software↗

A genome-wide survey of hybrid incompatibility factors by the introgression of marked segments of Drosophila mauritiana chromosomes into Drosophila simulans.

In hybrids between Drosophila simulans and D. mauritiana, males are sterile and females are fertile, in compliance with HALDANE's rule. The genetic basis of this phenomenon was investigated by introgression of segments of the mauritiana genome into a simulans background. A total of 87 positions throughout the mauritiana genome were marked with P-element insertions and replicate introgressions were made by repeated backcrossing to simulans for 15 generations. The fraction of hemizgyous X chromosomal introgressions that are male sterile is approximately 50% greater than the fraction of homozygous autosomal segments. This result suggests that male sterility factors have evolved at a higher rate on the X, but chromosomal differences in segment length cannot be ruled out. The fraction of homozygous autosomal introgression that are male sterile is several times greater than the fraction that are either female sterile or inviable. This observation strongly indicates that male sterility factors have evolved more rapidly than either female sterility or inviability factors. These results, combined with previous work on these and other species, suggest that HALDANE's rule has at least two causes: recessivity of incompatibility factors and differential accumulation of sterility factors affecting males and females.

Animals↗

Genetic architecture of species differences in annual sunflowers: implications for adaptive trait introgression.

Genetic architecture may profoundly influence the ability of adaptive traits to spread between species via introgressive hybridization. Here, we examine the genomic location of quantitative trait loci (QTL) associated with pollen sterility and morphological traits distinguishing two annual sunflowers, Helianthus annuus and H. debilis ssp. cucumerifolius. These species are of particular interest since they hybridize naturally, and the form of H. annuus in Texas (called ssp. texanus) is thought to have arisen through introgression. Analysis of 226 BC(1) progeny from a cross between H. annuus and H. debilis revealed 56 QTL for 15 morphological traits and 2 QTL for pollen sterility. Four morphological QTL are tightly linked (<10 cM) to one or more sterility factors and 7 are closely allied with underrepresented and presumably negatively selected chromosomal blocks. Although these 11 QTL seem unlikely to move between the species, no barrier to introgression was detected for the remaining 45 morphological QTL. In fact, due to widespread pleiotropy (or tight linkage), the introgression of just three small chromosomal blocks appears sufficient to largely recover the phenotype of ssp. texanus. Subsequent work will test for the occurrence and fitness consequences of the identified QTL in natural populations of ssp. texanus.

Base Sequence↗

Morphological, cytogenetic, and molecular evidence for introgressive hybridization in birch.

Extensive morphological variation of tetraploid birch (Betula pubescens) in Iceland is believed to be due to gene flow from diploid dwarf birch (B. nana) by means of introgressive hybridization. A combined morphological and cytogenetic approach was used to investigate this phenomenon in two geographically separated populations of natural birch woodland in Iceland. The results not only confirmed introgressive hybridization in birch, but also revealed bidirectional gene flow between the two species via triploid interspecific hybrids. The populations showed continuous morphological variation connecting the species, but karyotypically they consisted of only three types of plants: diploids, triploids, and tetraploids. No aneuploids were found. Some of the tetraploid plants had B. pubescens morphology as expected, but most of them had intermediate characters. Most of the diploid plants were B. nana, but some were intermediates and a few had B. pubescens morphology. The triploid plants were either intermediates or they resembled one of the two species. Similar introgressive variation was observed among the diploid and triploid progeny of open-pollinated B. nana in a garden. Birch samples including field plants and artificial hybrids were further examined using a molecular method based on genomic Southern hybridization. The experiments verified introgression at the DNA level.

Betula↗

Introgression of Luxilus cornutus mtDNA into allopatric populations of Luxilus chrysocephalus (Teleostei: Cyprinidae) in Missouri and Arkansas.

The cyprinid fishes Luxilus cornutus and Luxilus chrysocephalus hybridize extensively in a zone extending through the Great Lakes region with extensive introgression of L. cornutus mtDNA occurring in populations of L. chrysocephalus south of the present hybrid zone in Ohio. Western populations of these two species occur adjacent to one another in Missouri but hybridization has never been observed. In order to determine if hybridization has occurred historically in Missouri, allopatric populations of L. chrysocephalus were analysed for mtDNA introgression. Extensive introgression of L. cornutus mtDNA was observed in most populations of L. chrysocephalus in Missouri resulting in the elimination of L. chrysocephalus mtDNA in many populations. Luxilus cornutus mtDNA in L. chrysocephalus is found approximately 300 km south of extant L. cornutus populations in Missouri. Luxilus chrysocephalus mtDNA was replaced by four unique L. cornutus mtDNA haplotypes, with one particular haplotype becoming fixed in several L. chrysocephalus populations. The pattern of introgression suggests that historically L. cornutus occupied a more southern distribution in Missouri bringing it into contact with western populations of L. chrysocephalus and resulting in a hybrid zone.

Animals↗

Phylogeographic analysis of mitochondrial gene flow and introgression in the salamander, Plethodon shermani.

Plethodon shermani comprises a series of geographically disjunct populations occupying high-elevation mountain isolates. These populations hybridize at their borders with salamanders of the Plethodon glutinosus species complex, and past range expansions inferred from Pleistocene climatic cycles may have increased the possible genetic interactions between P. shermani and species of the P. glutinosus complex. Because mitochondrial DNA haplotypes often show introgression across species borders, we survey mtDNA variation for evidence of past and ongoing genetic interactions between P. shermani, its close relative Plethodon cheoah, and species of the P. glutinosus complex. Ongoing hybridization with the P. glutinosus-complex species Plethodon teyahalee is accompanied by extensive mitochondrial introgression in some Unicoi populations of P. shermani, but it has little genetic impact on P. shermani populations outside hybrid zones at three other isolates (Tusquitee, Wayah Bald, Standing Indian). Some Unicoi populations of P. shermani exhibit mtDNA evidence of past hybridization with diverse lineages from P. aureolus and P. glutinosus. The Tusquitee isolate of P. shermani is also characterized by mtDNA haplotypes most closely related to Plethodon aureolus and P. glutinosus, presumably introduced by past genetic contact with these species or with introgressed populations of Unicoi P. shermani. The mtDNA variation in sampled populations of the Wayah Bald and Standing Indian isolates of P. shermani appears largely unaffected by ongoing hybridization. Principal components analyses of allozymic data indicate that P. shermani isolates collectively form a genetically homogeneous unit clearly demarcated from species with which they have had current or past genetic interactions. Rapid mtDNA introgression associated with transient contacts between P. shermani and other species permits a fine-level resolution of evolutionary lineages not evident from allozymic data.

Animals↗