Separate Hybridization Events Research Articles

Calling Phenology and Call Structure of Sympatric Treefrogs in Eastern Texas

Anurans often call in acoustically complex choruses, which can consist of multiple closely related species. Closely related sympatric species often differ in phenology and the acoustic properties of male advertisement calls, which, in some species, are phenotypically plastic. The tetraploid Eastern Gray Treefrog (Hyla versicolor) originates from multiple, separate hybridization events of the diploid Cope's Gray Treefrog (Hyla chrysoscelis) with now-extinct diploid anurans. The two species are visually indistinguishable, exhibit highly similar ecology and behavior, and co-occur frequently across their range. Males of these species can be distinguished from each other by the structure of their advertisement calls. Daily audio recordings and weather data were collected for five consecutive years from eight breeding sites of sympatric H. versicolor and H. chrysoscelis. We examined spatial and temporal patterns in calling phenology and responses of both species to environmental cues. Additionally, we measured three advertisement call properties to evaluate differences in call structure based on community composition. The phenologies of the two gray treefrog species were nearly identical, and the presence of the other species did not significantly affect call pulse rate or dominant frequency in either species. Chorus composition affected call duration in both species, but in H. chrysoscelis, this effect was temperature dependent. Our results indicate that while calling phenology does not contribute to reproductive isolation in sympatric gray treefrogs, modulation of phenotypically plastic call properties could affect male mating success and prezygotic reproductive isolation.

A distinct and genetically diverse lineage of the hybrid fungal pathogen Verticillium longisporum population causes stem striping in British oilseed rape.

Population genetic structures illustrate evolutionary trajectories of organisms adapting to differential environmental conditions. Verticillium stem striping disease on oilseed rape was mainly observed in continental Europe, but has recently emerged in the United Kingdom. The disease is caused by the hybrid fungal species Verticillium longisporum that originates from at least three separate hybridization events, yet hybrids between Verticillium progenitor species A1 and D1 are mainly responsible for Verticillium stem striping. We reveal a hitherto un-described dichotomy within V. longisporum lineage A1/D1 that correlates with the geographic distribution of the isolates with an 'A1/D1 West' and an 'A1/D1 East' cluster. Genome comparison between representatives of the A1/D1 West and East clusters excluded population distinctiveness through separate hybridization events. Remarkably, the A1/D1 West population that is genetically more diverse than the entire A1/D1 East cluster caused the sudden emergence of Verticillium stem striping in the UK, whereas in continental Europe Verticillium stem striping is predominantly caused by the more genetically uniform A1/D1 East population. The observed genetic diversity of the A1/D1 West population argues against a recent introduction of the pathogen into the UK, but rather suggests that the pathogen previously established in the UK and remained latent or unnoticed as oilseed rape pathogen until recently.

Ranunculus schmalhausenii (section Batrachium, Ranunculaceae), a neglected water crowfoot endemic to Fennoscandia—a case of rapid hybrid speciation in postglacial environment of North Europe

The taxa of Ranunculus section Batrachium (Ranunculaceae) have been variably and unsatisfactorily treated in North Europe. Since the description of Ranunculus schmalhausenii (Batrachium dichotomum), probably the most common species in the area, its taxonomic status and identity have been unclear and differently implied. In the majority of treatments, individuals of R. schmalhausenii were ascribed to R. peltatus but sometimes also to the other morphologically similar, heterophyllous taxa. Based on detailed morphological study combined with geographical, ecological and biological evaluation the separate species status of this taxon was finally evidenced. The additive ITS polymorphism pattern of R. schmalhausenii confirmed its hybridogenous origin, however identification of the parental species was impeded by the heterogeneous character of the polymorphism detected. Genetic variation expressed by R. schmalhausenii samples may provide evidence of its multiple origin and suggests sexual reproduction of the taxon. Analysis of a sequence variation of two noncoding cpDNA regions, namely psbE-petL and rpl32-trnL, showed that individuals of R. schmalhausenii inherited cpDNA from two lineages of Batrachium, indicating that this taxon was created at least in two separate hybridization events. Ranunculus schmalhausenii may have originated from sexual ancestral species as multiple created hybrids which have been stabilized by polyploidisation. Genetic structure of R. schmalhausenii is somehow similar to also hybridogenous R. penicillatus. In this study, a detailed morphological, geographical, ecological, and biological description of R. schmalhausenii was presented and the differences between this species and similar taxa were outlined. The name was lectotypified and its synonymy was provided. In contrast to many other heterophyllous species of Ranunculus section Batrachium, R. schmalhausenii occurs mainly in young, postglacial landscapes of Fennoscandia, prefering deep and clear waters with current or wave action and a hard bottom, which perfectly corresponds with a relict, postglacial nature of the species. The species probably presents an example of rapid hybrid speciation (less than 10 000 years) in postglacial environment of North Europe and may be considered as endemic to Fennoscandia. Moreover, R. schmalhausenii, as a weak competitor and pollution sensitive taxon, can be regarded as an indicator of clean waters. Phylogenetic relations within section Batrachium indicates convergent evolution of some species and two cases of possible cpDNA capture.

A large set of newly created interspecific Saccharomyces hybrids increases aromatic diversity in lager beers.

Lager beer is the most consumed alcoholic beverage in the world. Its production process is marked by a fermentation conducted at low (8 to 15°C) temperatures and by the use of Saccharomyces pastorianus, an interspecific hybrid between Saccharomyces cerevisiae and the cold-tolerant Saccharomyces eubayanus. Recent whole-genome-sequencing efforts revealed that the currently available lager yeasts belong to one of only two archetypes, "Saaz" and "Frohberg." This limited genetic variation likely reflects that all lager yeasts descend from only two separate interspecific hybridization events, which may also explain the relatively limited aromatic diversity between the available lager beer yeasts compared to, for example, wine and ale beer yeasts. In this study, 31 novel interspecific yeast hybrids were developed, resulting from large-scale robot-assisted selection and breeding between carefully selected strains of S. cerevisiae (six strains) and S. eubayanus (two strains). Interestingly, many of the resulting hybrids showed a broader temperature tolerance than their parental strains and reference S. pastorianus yeasts. Moreover, they combined a high fermentation capacity with a desirable aroma profile in laboratory-scale lager beer fermentations, thereby successfully enriching the currently available lager yeast biodiversity. Pilot-scale trials further confirmed the industrial potential of these hybrids and identified one strain, hybrid H29, which combines a fast fermentation, high attenuation, and the production of a complex, desirable fruity aroma.

Loss of lager specific genes and subtelomeric regions define two different Saccharomyces cerevisiae lineages for Saccharomyces pastorianus Group I and II strains

Lager yeasts, Saccharomyces pastorianus, are interspecies hybrids between S. cerevisiae and S. eubayanus and are classified into Group I and Group II clades. The genome of the Group II strain, Weihenstephan 34/70, contains eight so-called 'lager-specific' genes that are located in subtelomeric regions. We evaluated the origins of these genes through bioinformatic and PCR analyses of Saccharomyces genomes. We determined that four are of cerevisiae origin while four originate from S. eubayanus. The Group I yeasts contain all four S. eubayanus genes but individual strains contain only a subset of the cerevisiae genes. We identified S. cerevisiae strains that contain all four cerevisiae 'lager-specific' genes, and distinct patterns of loss of these genes in other strains. Analysis of the subtelomeric regions uncovered patterns of loss in different S. cerevisiae strains. We identify two classes of S. cerevisiae strains: ale yeasts (Foster O) and stout yeasts with patterns of 'lager-specific' genes and subtelomeric regions identical to Group I and II S. pastorianus yeasts, respectively. These findings lead us to propose that Group I and II S. pastorianus strains originate from separate hybridization events involving different S. cerevisiae lineages. Using the combined bioinformatic and PCR data, we describe a potential classification map for industrial yeasts.

Polyphyletic origin of Brassica juncea with B. rapa and B. nigra (Brassicaceae) participating as cytoplasm donor parents in independent hybridization events.

• Premise of the study: Brassica juncea is a major source of edible oil in the Indian subcontinent and northern China. It is also used as a root and leaf vegetable in China and as a condiment in Europe and America. There is a long-standing view that B. juncea originated from multiple hybridization events between B. rapa and B. nigra and that hybridizations were always unidirectional with B. rapa as the cytoplasmic donor. These conclusions were, however, centered primarily on nuclear markers.• Methods: Two hundred forty-six accessions of B. juncea, B. rapa, and B. nigra were genotyped using chloroplast and nuclear simple sequence repeat (SSR) markers.• Key results: A structure analysis assigned B. juncea germplasm (122) into three major groups based on plasmotype variation. The bulk of Indian B. juncea genotypes were grouped along with Chinese and Australian accessions. This plasmotype was absent in sampled accessions of B. rapa (97), B. nigra (27), and other wild crucifers (10). The second group of B. juncea included East European genotypes and four accessions from India. It showed unambiguous homology with the predominant B. nigra plasmotype. The neighbor joining tree produced seven subgroups, arranged into two broad lineages. The first lineage included Indian, Australian, and Chinese B. juncea genotypes; it was associated with wild species belonging to the "rapa" lineage. Nuclear SSR marker-based analyses were largely supportive of results from chloroplast SSR analyses.• Conclusions: Based on these results, we provide the first report that B. juncea originated several times with both B. rapa and B. nigra as cytoplasmic donors in separate hybridization events.

Investigating temporal changes in hybridization and introgression in a predominantly bimodal hybridizing population of invasive sika (Cervus nippon) and native red deer (C. elaphus) on the Kintyre Peninsula, Scotland

We investigated temporal changes in hybridization and introgression between native red deer (Cervus elaphus) and invasive Japanese sika (Cervus nippon) on the Kintyre Peninsula, Scotland, over 15 years, through analysis of 1513 samples of deer at 20 microsatellite loci and a mtDNA marker. We found no evidence that either the proportion of recent hybrids, or the levels of introgression had changed over the study period. Nevertheless, in one population where the two species have been in contact since approximately 1970, 44% of individuals sampled during the study were hybrids. This suggests that hybridization between these species can proceed fairly rapidly. By analysing the number of alleles that have introgressed from polymorphic red deer into the genetically homogenous sika population, we reconstructed the haplotypes of red deer alleles introduced by backcrossing. Five separate hybridization events could account for all the recently hybridized sika-like individuals found across a large section of the Peninsula. Although we demonstrate that low rates of F1 hybridization can lead to substantial introgression, the progress of hybridization and introgression appears to be unpredictable over the short timescales.

Phylogeography of sexualHeteronotia binoei(Gekkonidae) in the Australian arid zone: climatic cycling and repetitive hybridization

The biota of much of continental Australia have evolved within the context of gradual aridification of the region over several million years, and more recently of climatic cycling between relatively dry and humid conditions. We performed a phylogeographical study of three sexual chromosome races of the Heteronotia binoei complex of geckos found throughout the Australian arid zone. Two of these three races were involved in two separate hybridization events leading to parthenogenetic lineages (also H. binoei), and the third is widespread and broadly sympatric with the parthenogens. Based on our analyses, the three sexual races diversified approximately 6 million years ago in eastern Australia, during a period of aridification, then each moved west through northern, southern, and central dispersal corridors to occupy their current ranges. In each case, the timing of major phylogeographical inferences corresponds to inferred palaeoclimatic changes in continental Australia. This scenario provides a simple explanation for diversification, secondary contact, and hybridization between the races. However, data presented elsewhere indicate that formation of the parthenogens was considerably more recent than the westward expansion of the hybridizing races, and that multiple hybridization events were geographically and temporally distinct. We suggest that cyclical climate changes may have led to regional range changes that facilitated hybridization between the races, which are not currently known to be in sympatry.

Isolates of Verticillium dahliae Pathogenic to Crucifers Are of at Least Three Distinct Molecular Types

ABSTRACT Diverse isolates of the soilborne wilt fungi Verticillium dahliae and V. albo-atrum were studied to understand the nature and origins of those infecting cruciferous hosts. All isolates from cruciferous crops produced microsclerotia, and the majority produced long conidia with a high nuclear DNA content; these isolates were divided into two groups by amplified fragment length polymorphism (AFLP) analysis. One group could be subdivided by other criteria such as rRNA sequences and mitochondrial DNA restriction fragment length polymorphism (RFLP) analysis. Two crucifer isolates were short spored and had a low nuclear DNA content. The results are consistent with the crucifer isolates being interspecific hybrids. The long-spored isolates are best regarded as amphihaploids (or allodiploids) with the AFLP groups probably each representing separate interspecific hybridization events. The short-spored crucifer isolates appear to be derived from interspecific hybrids and are here called 'secondary haploids'. Molecular evidence suggests that one parent in the crosses was similar to V. dahliae. The other parent of the amphihaploids seems to have been more similar to V. albo-atrum than to V. dahliae, but was distinct from all isolates of either species so far studied. The implications for the taxonomy of crucifer isolates are discussed and the use of the name V. longisporum, proposed elsewhere for just some of these isolates, is discouraged.

Divergence of the CytochromebGene in theLacerta raddeiComplex and Its Parthenogenetic Daughter Species: Evidence for Recent Multiple Origins

Abstract Questions concerning the origin of parthenogenesis in Caucasian Rock Lizards and genetic divergence among bisexual lizards of the Lacerta raddei complex were examined using sequences from the mitochondrial cytochrome b gene. The maternal parent of the parthenogenetic L. uzzelli, L. sapphirina, and L. bendimahiensis was confirmed to be L. raddei. Although substantial variation was revealed among bisexual populations of L. raddei and L. nairensis, very low or no variation was found among the parthenogenetic species. A phylogenetic tree including 11 populations of L. raddei and L. nairensis, as well as 10 populations of its five daughter parthenogens, was constructed. Because of paraphyletic relationships, L. nairensis is considered conspecific with L. raddei. Evaluation of the parthenogenetic species suggests that separate hybridization events between L. raddei and L. valentini might have occurred at least twice. One resulted in L. sapphirina and L. bendimahiensis, and the other one (or more) resul...

Allopolyploid origin and population genetics of the rare orchid Spiranthes diluvialis

The process of becoming and the attributes of being polyploid play a major role in the development and maintenance of genetic variation in allopolyploid species. A genetic survey employing protein electrophoresis on 12 populations of S. diluvialis, as well as on populations of eight congeneric species, was conducted to assess the putative allopolyploid origin of S. diluvialis and to determine the genetic variability within and among populations. Genetic identity values indicated S. diluvialis was more similar to S. magnicamporum (0.619) and S. romanzoffiana (0.727) than to any of the other congeneric species assayed. Similar to most allopolyploids, S. diluvialis showed high levels of fixed, or nearly fixed, heterozygosity and a high percentage of polymorphic loci (57.1-71.4%). The mean number of alleles per polymorphic locus in populations of S. diluvialis (2.6-3.3), however, was similar to mean values for both animal-pollinated, outcrossing, diploid species, and geographically restricted, diploid species (2.6 and 2.5, respectively). Genetic divergence among populations (mean Fst = 0.083) was low, leading to relatively high estimates of interpopulational gene flow (mean Nm = 5.41). Thus, each population harbors most of the genetic variability found within the species. The genetic variation observed within S. diluvialis supports the occurrence of at least two separate hybridization events giving rise to S. diluvialis.

Chromosome substitutions of Triticum timopheevii in common wheat and some observations on the evolution of polyploid wheat species

Whether the two tetraploid wheat species, the well known Triticum turgidum L. (macaroni wheat, AABB genomes) and the obscure T. timopheevii Zhuk. (A(t)A(t)GG), have monophyletic or diphyletic origin from the same or different diploid species presents an interesting evolutionary problem. Moreover, T. timopheevii and its wild form T. araraticum are an important genetic resource for macaroni and bread-wheat improvement. To study these objectives, the substitution and genetic compensation abilities of individual T. timopheevii chromosomes for missing chromosomes of T. aestivum 'Chinese Spring' (AABBDD) were analyzed. 'Chinese Spring' aneuploids (nullisomic-tetrasomics) were crossed with a T. timopheevii x Aegilops tauschii amphiploid to isolate T. timopheevii chromosomes in a monosomic condition. The F1 hybrids were backcrossed one to four times to Chinese Spring aneuploids without selection for the T. timopheevii chromosome of interest. While spontaneous substitutions involving all A(t)- and G-genome chromosomes were identified, the targeted T. timopheevii chromosome was not always recovered. Lines with spontaneous substitutions from T. timopheevii were chosen for further backcrossing. Six T. timopheevii chromosome substitutions were isolated: 6A(t) (6A), 2G (2B), 3G (3B), 4G (4B), 5G (5B) and 6G (6B). The substitution lines had normal morphology and fertility. The 6A(t) of T. timopheevii was involved in a translocation with chromosome 1G, resulting in the transfer of the group-1 gliadin locus to 6A(t). Chromosome 2G substituted for 2B at a frequency higher than expected and may carry putative homoeoalleles of gametocidal genes present on group-2 chromosomes of several alien species. Our data indicate a common origin for tetraploid wheat species, but from separate hybridization events because of the presence of a different spectrum of intergenomic translocations.

Isozyme Evidence of Reticulate Evolution in Mosses: Plagiomnium medium is an Allopolyploid of P. ellipticum x P. insigne

Electrophoretic data from 44 natural populations show that Plagiomnium medium is an allopolyploid, whose genome can be reconstructed by merely combining present-day variation of its extant haploid progenitors, P. ellipticum and P. insigne. Because the progenitor species are well- differentiated genetically, P. medium shows fixed heterozygosity in about 1/3 of the 23 loci assayed. High levels of polymorphism in P. ellipticum provide the opportunity to detect multiple origins of P. medium, which evidence shows must have originated from at least four separate hybridization events. We detected only one allele unique to P. medium and only one possible instance of gene silencing. This suggests a relatively recent origin of the allopolyploid. It may also suggest that natural selection does not favor silencing back to a haploid condition. Apparently, cross-fertilization is frequent in P. medium and generates a large number of genetic recombinants across loci. Thus, P. medium enjoys a favorable genetic situation in which there is fixed, intralocus heterozygosity combined with the potential for extensive interlocus recombination. We speculate that P. medium may have been launched as a species during the Pleistocene, when the ranges of the circumboreal, continental P. ellipticum and the coastal, Pacific Northwest endemic P. insigne may have overlapped broadly. Their present-day ranges include few sympatric populations and would not permit hy- bridization along a broad front. Contrary to conventional views, it appears that reticulate evolution occurs in bryophytes, just as it does in other land plants.

Assignment of the genes for mouse type I procollagen to chromosome 16 using mouse fibroblast-Chinese hamster somatic cell hybrids.

Somatic cell hybrids between mouse and Chinese hamster fibroblasts have been used to identify the chromosome responsible for the synthesis of both mouse type I procollagen subunit chains (MCOLA1 and MCOLA2). Thirty-one separate hybrid clones and subclones from ten separate hybridization events were isolated in hypoxanthine-aminopterin-thymidine (HAT) selection medium and were used for detailed gene-mapping studies. ELISA and "Western blotting" immunochemical analysis were used to detect the production of mouse type I procollagen in each hybrid clone. Mouse and Chinese hamster chromosomes were identified in each hybrid clone by trypsin-Giemsa banding of metaphase chromosome spreads and by isozyme analysis. We have found that mouse type I procollagen production segregates concordantly with mouse superoxide dismutase-1, previously mapped to mouse chromosome 16, and with the presence of mouse chromosome 16 karyotypically. Western blotting immunochemical analysis of the separated mouse procollagen chains produced by each hybrid line demonstrated that apparently the genes for both subunit chains are located on the same chromosome. These studies, therefore, assign the structural genes for mouse type I procollagen pro alpha 1 (MCOLA1) and pro alpha 2 (MCOLA2) chains to mouse chromosome 16.