The influence of hybridization on epidermal properties of birch species and the consequences for palaeoclimatic interpretations

The Fennoscandian birch population primarily consists of Betula nana, B. pendula and B. pubescens ssp. czerepanovii, the Mountain birch. Frequent hybridization between the Mountain birch and B. nana generates a wide range of genotypic and phenotypic plasticity in the subarctic birch zone of Fennoscandia. Phases of subarctic conditions prevailed during the Late Glacial in large parts of NW Europe, and palynological as well as macrofossil analysis provide some evidence for the occurrence of birch hybrids during these intervals. Leaves from genetically controlled specimens of Betula pendula, B. pubescens ssp. czerepanovii, B. nana and the hybrids B. pubescens ssp. czerepanovii × nana and B. nana × pubescens ssp. czerepanovii are investigated for their specific characteristics of the epidermis morphology. Frequency and size of epidermal cells and stomata reveal a close affinity of both hybrids to B. nana and allow a differentiation of the intermediate forms between B. nana and the Mountain birch. With respect to palaeoatmospheric CO₂ reconstructions based on stomatal index, epidermal analysis shows that a possible occurrence of hybrids in fossil leaf assemblages has no profound consequences for combined species records. However, the significant differences observed in B. nana demand the separation of this species. A comparison of the cuticle properties of B. pendula and B. pubescens from Finnish Lapland and leaf material from The Netherlands reveals a divergence of the stomatal index that may be due to differences in day light length.     

Comparative analyses of plastid and AFLP data suggest different colonization history and asymmetric hybridization between Betula pubescens and B. nana

Birches (Betula spp.) hybridize readily, confounding genetic signatures of refugial isolation and postglacial migration. We aimed to distinguish hybridization from range‐shift processes in the two widespread and cold‐adapted species B. nana and B. pubescens, previously shown to share a similarly east–west‐structured variation in plastid DNA (pDNA). We sampled the two species throughout their ranges and included reference samples of five other Betula species and putative hybrids. We analysed 901 individual plants using mainly nuclear high‐resolution markers (amplified fragment length polymorphisms; AFLPs); a subset of 64 plants was also sequenced for two pDNA regions. Whereas the pDNA variation as expected was largely shared between B. nana and B. pubescens, the two species were distinctly differentiated at AFLP loci. In B. nana, both the AFLP and pDNA results corroborated the former pDNA‐based hypothesis that it expanded from at least two major refugia in Eurasia, one south of and one east of the North European ice sheets. In contrast, B. pubescens showed a striking lack of geographic structuring of its AFLP variation. We identified a weak but significant increase in nuclear (AFLP) gene flow from B. nana into B. pubescens with increasing latitude, suggesting hybridization has been most frequent at the postglacial expansion front of B. pubescens and that hybrids mainly backcrossed to B. pubescens. Incongruence between pDNA and AFLP variation in B. pubescens can be explained by efficient expansion from a single large refugium combined with leading‐edge hybridization and plastid capture from B. nana during colonization of new territory already occupied by this more cold‐tolerant species.     

Multispecies genetic structure and hybridization in the Betula genus across Eurasia

Boreal and cool temperate forests are the major land cover of northern Eurasia, and information about continental‐scale genetic structure and past demographic history of forest species is important from an evolutionary perspective and has conservation implications. However, although many population genetic studies of forest tree species have been conducted in Europe or Eastern Asia, continental‐scale genetic structure and past demographic history remain poorly known. Here, we focus on the birch genus Betula, which is commonly distributed in boreal and cool temperate forests, and examine 129 populations of two tetraploid and four diploid species collected from Iceland to Japan. All individuals were genotyped at seven to 18 nuclear simple sequence repeats (nSSRs). Pairwise among the six species ranged from 0.285 to 0.903, and genetic differentiation among them was clear. structure analysis suggested that Betula pubescens is an allotetraploid and one of the parental species was Betula pendula. In both species pairs of B. pendula and B. plathyphylla, and B. pubescens and B. ermanii, genetic diversity was highest in central Siberia. A hybrid zone was detected around Lake Baikal for eastern and western species pairs regardless of ploidy level. Approximate Bayesian computation suggested that the divergence of B. pendula and B. platyphylla occurred around the beginning of the last ice age (36 300 years BP, 95% CI: 15 330–92 700) and hybridization between them was inferred to have occurred after the last glacial maximum (1614 years BP, 95% CI: 561–4710), with B. pendula providing a higher contribution to hybrids.     

Essential oil in Betula spp. leaves naturally growing in Estonia

The essential oils of the leaves of Betula pendula Roth., Betula pubescens Ehrh., Betula humilis Schrank and Betula nana L. (Betulaceae), and B. pendula buds were obtained using hydrodistillation and their chemical compositions were analyzed using gas chromatography and gas chromatography – mass spectrometry. The yield of essential oil was the highest (0.27%) in leaves of. B. humilis and the lowest in the leaves of B. pendula (0.11%), B. pubescens (0.05%) and B. nana (traces). Overall, 35–60 compounds were identified representing more than 92% of the total volume of the oils investigated. The bicyclical sesquiterpenoids found in the essential oils extracted from the leaves of B. pendula, B. pubescens and B. humilis and from the buds of B. pendula where α-betulenol (20.4–33.1%), α-betulenol acetate (5,6–28.9%), β-betulenal (5.5–6.2%) and β-betulenol (2.0–5.8%). The buds of B. pendula contained more birkenal, (E)-β-caryophyllene, δ-cadinene and caryophyllene oxide, but less α-betulenol acetate, β-betulenal and aliphatic compounds than the oils extracted from the leaves investigated. The oil composition of B. humilis and B. nana leaves was studied for the first time. The content of essential oil of B. nana leaves was much more varied: the oil contained less bicyclical sesquiterpenoids and more aliphatic compounds than the other Betula species investigated. The findings of our study could be used in chemosystematics of birch species, especially for B. nana.     

Differentiating pollen of Betula species from Iceland

Subfossil pollen from two co‐existing Betula species in Iceland, B. nana and B. pubescens, is frequently found in sediments and peat. Interpretation of the findings often depends on the ability to differentiate between the two species according to pollen size and structure. Fresh pollen samples were prepared from 70 individual trees/shrubs which had been identified to species by chromosome number. Grain diameters and pore depths were measured and ratios of grain diameter to pore depth (D/P ratios) were calculated. The mean grain diameters of pollen from diploid B. nana and tetraploid B. pubescens were 20.42 and 24.20?µm, whereas mean pore depths were 2.20 and 2.81?µm respectively. Mean D/P ratios were therefore 9.55 for B. nana and 8.85 for B. pubescens. The difference between species was statistically significant for all three pollen parameters. Grain diameter appeared to be the most useful parameter, as only about 20% of the samples were in the overlapping region of the species distributions. Pollen size (grain diameter) was also positively correlated to tree morphology, which was evaluated using species‐specific botanical characters. Pollen samples from different locations/populations in Iceland varied slightly in mean size and ratio. The size difference between pollen of B. nana and B. pubescens in this study is less than other papers have reported, which may be due to the effect of introgressive hybridisation between the two birch species in Iceland.     

The identification of wingless Betula fruits in Weichselian sediments in the Gross Todtshorn borehole (Lower Saxony, Germany) – the occurrence of Betula humilis Schrank.

Betula fruits lacking their wings were recovered from the Gross Todtshorn borehole, Germany, covering the late Eemian interglacial, the Early Weichselian, and the Weichselian pleniglacial. In order to identify them, a data-set of seven size and shape variables was made from modern fruits of B. pubescens, B. pendula, B. nana, and B. humilis. These measurements were summarised by principal components analysis (PCA). The 140 measured fossil fruit-bodies were positioned passively on the PCA plot and identified by comparison with the dimensional envelopes of the modern species. Tree birches (B. pendula, B. pubescens) occurred in the Eemian and Early Weichselian interstadials, but not thereafter.B. nana was widespread during the intervening stadials and the Oerel interstadial. B. humilis occurred throughout the sequence, coexisting with tree birches in the Early Weichselian and with B. nana in the stadials. It usually grew on mire surfaces, in contrast to B. nana which mainly grew on mineral substrates. However, B. nana grew on peat with B. humilis in the pleniglacial Oerel interstadial, suggesting a more continental climate with cold winters. A modern analogue could be near the Ural mountains. B. humilis is presently rare in central Europe, but during the Weichselian it was well established in northern Germany. The PCA has enabled specific determination of the wingless Betula fruits, leading to vegetational and climatic reconstructions and contributing to the previously poorly known historical biogeography of Betula humilis. Received June 14, 2000 / Accepted January 2, 2001     

Natural hybridisation in birch: triploid hybrids between Betula nana and B. pubescens

Hybridisation between diploid (2n=28) dwarf birch Betula nana L. and tetraploid (2n=56) downy birch B. pubescens Ehrh. has occurred in natural populations in Iceland. About 10% of birch plants randomly collected are triploid (2n=42) hybrids. Ribosomal gene mapping on chromosomes and genomic in situ hybridisation confirms the hybridity. However, the triploid hybrids are not morphologically distinct, i.e. they are not different from diploid and tetraploid birch plants that have intermediate morphology. The triploid hybrids have evidently played an important role in driving bi-directional gene flow between these two species. This paper reviews the extent of interspecific hybridisation in selected birch woodland populations and discusses the significance of natural hybridisation and introgression in birch.     

Introgression and phylogeography of Betula nana (diploid), B. pubescens (tetraploid) and their triploid hybrids in Iceland inferred from cpDNA haplotype variation

Aim The objective was to find direct genetic evidence supporting introgressive hybridization between tetraploid tree birch (Betula pubescens) and diploid dwarf birch (B. nana), via triploid hybrids, and to investigate an association between the introgression and phylogeographical distribution of Icelandic birch. Location Samples were collected from 463 trees in 12 woodlands in Iceland and eight locations in Norway, Sweden, Scotland and Greenland. Methods Ploidy status of individual trees was determined by chromosome counting. Variation in the chloroplast genome was assessed using polymerase chain reaction‐restriction fragment length polymorphism. The geographical distribution of the haplotypes was mapped. The haplotype variation and introgression ratios (IG) were analysed statistically. Results Thirteen haplotypes were identified among Icelandic samples. The most common haplotype (T, 49% occurrence) was present in all ploidy groups and in all woodlands. All common haplotypes were shared between the triploid group and the parental species, indicating introgressive hybridization. This was supported by the statistical analysis of IG indices and the variation components. Considerable differences existed among samples, shaped by isolation by distance and local introgression. An east–west phylogeographical distribution in Iceland was observed. Main conclusions Despite extensive introgression across species and ploidy levels, a biogeographical pattern has been observed, and this may indicate different population histories or multiple origins of Icelandic birch. The chloroplast haplotype diversity found in Iceland resembles that found in birch populations from northern Scandinavia.     

Characteristics of pollen from natural triploid Betula hybrids

Birch has a key role in the Holocene vegetation history of northern Europe and in sub‐arctic climates dwarf birch and tree birch co‐exist. In Iceland, triploid hybrids between diploid Betula nana (dwarf birch) and tetraploid B. pubescens (downy birch) are common and therefore likely to contribute to pollen deposition. Pollen from 22 triploid trees/shrubs from ten woodlands in Iceland was examined and its size and shape compared with pollen from the parental species. The mean diameter of pollen grains from the triploid hybrids was not statistically different from that of B. nana pollen, but was significantly smaller than the mean value of B. pubescens pollen. On the contrary, the size of the vestibulum was similar to that of B. pubescens, which was significantly greater than that of B. nana, and therefore the diameter‐pore depth ratio was lower than the values from either species. The pattern of size distribution within plants indicated that triploid hybrids might have produced two sizes of triporate pollen grains, but the small B. nana size was far more prevalent than the larger B. pubescens size. Several anomalies in pollen morphology were common among the hybrid pollen grains: four or more pores were the most frequent type of abnormality. Characteristics of the pollen of triploid Betula hybrids, especially structural anomalies, may provide a means to reveal periods of interspecific hybridisation in the analysis of sub‐fossil pollen.     

Size of the leaf as a marker of birch productivity at a distance from the climatic optimum

Morphometric parameters of mature leaves (area, venation density, and shape coefficient) and trunks (size and biomass) of two edificatory birch species, Betula pendula Roth and B. pubescens Ehrh., were characterized. These species inhabit a 1600-km transect of the Urals, from forest-tundra to forest-steppe. In both species, the highest trunk biomass was observed in the subzone of south taiga. Trunk parameters and leaf sizes of B. pendula varied in a narrower range than in B. pubescens. As distinct from leaf shape, leaf size was correlated with the average multiyear climate characteristics but not with the weather during a given growth season. In B. pubescens, which range extends wide from its climatic optimum, leaf area was positively correlated with trunk biomass. In B. pendula, growing mainly within the area of its climatic optimum, such correlation was not observed. We concluded that such parameter as mature leaf size could be used for prediction of tree productivity growing outside of the zone of their climatic optimum.     

Bidirectional mitochondrial introgression between Korean cobitid fish mediated by hybridogenetic hybrids

Genomic introgression through interspecific hybridization has been observed in some species of the freshwater fish family Cobitidae. Within this family, a Cobitis hankugensis–Iksookimia longicorpa diploid–triploid hybrid species complex on the Korean peninsula is unique in displaying hybridogenesis, a unisexual reproduction mode that allows hybrids to mediate the transfer of mitochondrial DNA (but not nuclear DNA) between the two parent species. However, populations of the parental species in the wild have never been examined for the potential effect of introgression on their genomes. To address the genetic consequences of unisexual hybridization on the parental species, we examined genetic structure of the two parental species, C. hankugensis and I. longicorpa, in three independent natural habitats where they coexist with their hybrid complex using DNA sequence data of one mitochondrial gene and three nuclear genes. We found that mitochondrial introgression between the two species was extensive in all the examined localities, while there was no evidence of nuclear introgression across the species boundary. This result indicates that the hybridogenetic individuals mediate mitochondrial introgression from one species to the other, producing mito‐nuclear mosaic genomes such as C. hankugensis nuclear genomes associated with I. longicorpa mitochondrial DNA and the reverse. The direction and degree of introgression varied among the three localities, but the underlying mechanisms for this observation proved elusive. Introgression might depend on which species serves as the predominant sperm or ovum donor or the environmental conditions of the localities. The present study suggests that introgressive hybridization between pure C. hankugensis and I. longicorpa species is highly likely where the two species co‐occur with hybridogenetic individuals, but the consequence of introgression could be variable due to the history and environmental characteristics of particular populations across the parental species’ ranges.     

Massive invasion of exotic Barbus barbus and introgressive hybridization with endemic Barbus plebejus in Northern Italy: where,how and why?

Biological invasions and introgressive hybridization are major drivers for the decline of native freshwater fish. However, the magnitude of the problem across a native species range, the mechanisms shaping introgression as well as invader's dispersal and the relative role of biological invasions in the light of multiple environmental stressors are rarely described. Here, we report extensive (N = 665) mtDNA sequence and (N = 692) microsatellite genotypic data of 32 Northern Adriatic sites aimed to unravel the invasion of the European Barbus barbus in Italy and the hybridization and decline of the endemic B. plebejus. We highlight an exceptionally fast breakthrough of B. barbus within the Po River basin, leading to widespread introgressive hybridization with the endemic B. plebejus within few generations. In contrast, adjacent drainage systems are still unaffected from B. barbus invasion. We show that barriers to migration are inefficient to halt the invasion process and that propagule pressure, and not environmental quality, is the major driver responsible for B. barbus success. Both introgressive hybridization and invader's dispersal are facilitated by ongoing fisheries management practices. Therefore, immediate changes in fisheries management (i.e. stocking and translocation measures) and a detailed conservation plan, focussed on remnant purebred B. plebejus populations, are urgently needed.     

Distinguishing fossil Betula nana and B. pubescens using their wingless fruits: implications for the late-glacial vegetational history of western Norway

Principal components analysis of 7 size and shape variables measured on 154 modern fruit bodies of Betula species and hybrids shows that B. pubescens and B. nana can be distinguished from each other on the basis of their fruit bodies, and that B. pubescens ssp. tortuosa and hybrids either fall within the range of B. pubescens or have an intermediate position on PCA axes 1 and 2. Passive positioning on the modern PCA axes of fossil fruit bodies from late-glacial sediments at Eigebakken, south-west Norway, shows that only B. nana was locally present in the Allerød. This contradicts earlier inferences from pollen analyses alone of birch woodland development in the Allerød in south-west Norway. There is no macrofossil evidence for tree-birches in the Allerød near Eigebakken. The relatively large amounts of Betula pollen, including B. pubescens, at Eigebakken are thus probably derived by long-distance transport from tree-birches in Denmark, south Sweden, and Britain, whose presence is proved there by macrofossils. Consequently, earlier estimates of mean July temperature during the Allerød in southwestern Norway should be reduced to around 7.5–10°C. In contrast, the PCA shows that fruit bodies from Holocene sediments at Eigebakken and at Kråkenes, western Norway, are mostly derived from B. pubescens. Birch trees were able to spread quickly and effectively across Norway in the early Holocene.     

A Lichenological Excursion to the West of Scotland

ABSTRACT

Background: Hybridisation can be a threat for the survival of a rare species because, in the case of insufficient numbers of appropriate mates, a rare form is much more likely to cross with a widespread taxon.

Aims: In the present study, we tested hypotheses concerning the level of hybridisation between endangered Betula humilis and its widespread congeners: B. pendula and B. pubescens as a function of habitat conditions.

Methods: We genotyped 312 individuals of three species using AFLP markers. B. humilis specimens were sampled in populations with low and high groundwater levels. Morphological identification of B. pubescens and B. pendula was verified using the Atkinson discriminant function.

Results: Altogether, 15 individuals (4.8%) were indicated as putative hybrids. The B. humilis hybrids were found in dry habitats and they could be classified as F1 or F2 generation. Tree hybrids could represent backcrosses to either B. pendula or B. pubescens.

Conclusions: Genetic analyses contradicted the idea that hybridisation between B. humilis and its close relatives was extensive. On the other hand, the presence of introgressed individuals in the populations in areas with low groundwater levels implied that pollen swamping might be a threat for declining B. humilis stands.     

Molecular identification of natural hybridization between Melastoma malabathricum and Melastoama beccarianum in Sarawak,Malaysia

Hybridization is very common in flowering plants and it plays a significant role in plant evolution and adaptation. Melastoma L. (Melastomataceae) comprises about 80–90 species in tropical Asia and Oceania, among which 41 species occur in Borneo. Natural hybridization is frequently reported in Melastoma in China, but so far there have been no confirmed cases of hybridization in Southeast Asia (including Borneo), where most species occur. Here, we identified a case of natural hybridization between Melastoma malabathricum L. and Melastoma beccarianum Cogn. in Sarawak, Malaysia, by using sequence data of three nuclear genes and one chloroplast intergenic spacer. Melastoma malabathricum is the most widespread species of this genus, occurring in almost the whole range of this genus, while M. beccarianum is a local species endemic to northern Borneo. Our results showed that natural hybridization and introgression occur between M. malabathricum and M. beccarianum, and the introgression was asymmetrical, mainly from M. malabathricum to M. beccarianum. As adaptive traits can be transferred by introgression, our study suggests that natural hybridization should be a significant mechanism for the evolution and adaptation of Melastoma in Southeast Asia. However, introgression from the common species M. malabathricum to the relatively rare species M. beccarianum may cause the decline of M. beccarianum, incurring conservation concern. With a large number of species of Melastoma and almost year‐around flowering in Southeast Asia, more cases of natural hybridization are expected to be found and identified in near future.     

Sequential adaptive introgression of the mitochondrial genome in Drosophila yakuba and Drosophila santomea

Interspecific hybridization provides the unique opportunity for species to tap into genetic variation present in a closely related species and potentially take advantage of beneficial alleles. It has become increasingly clear that when hybridization occurs, mitochondrial DNA (mtDNA) often crosses species boundaries, raising the possibility that it could serve as a recurrent target of natural selection and source of species' adaptations. Here we report the sequences of 46 complete mitochondrial genomes of Drosophila yakuba and Drosophila santomea, two sister species known to produce hybrids in nature (~3%). At least two independent events of mtDNA introgression are uncovered in this study, including an early invasion of the D. yakuba mitochondrial genome that fully replaced the D. santomea mtDNA native haplotypes and a more recent, ongoing event centred in the hybrid zone. Interestingly, this recent introgression event bears the signature of Darwinian natural selection, and the selective haplotype can be found at low frequency in Africa mainland populations of D. yakuba. We put forward the possibility that, because the effective population size of D. santomea is smaller than that of D. yakuba, the faster accumulation of mildly deleterious mutations associated with Muller's ratchet in the former species may have facilitated the replacement of the mutationally loaded mitochondrial genome of D. santomea by that of D. yakuba.     

Fine‐scale geographic patterns of gene flow and reproductive character displacement in Drosophila subquinaria and Drosophila recens

When two species are incompletely isolated, strengthening premating isolation barriers in response to the production of low fitness hybrids may complete the speciation process. Here, we use the sister species Drosophila subquinaria and Drosophila recens to study the conditions under which this reinforcement of species boundaries occurs in natural populations. We first extend the region of known sympatry between these species, and then we conduct a fine‐scale geographic survey of mate discrimination coupled with estimates of gene flow within and admixture between species. Within D. subquinaria, reinforcement is extremely effective: we find variation in mate discrimination both against D. recens males and against conspecific allopatric males on the scale of a few kilometres and in the face of gene flow both from conspecific populations and introgression from D. recens. In D. recens, we do not find evidence for increased mate discrimination in sympatry, even where D. recens is rare, consistent with substantial gene flow throughout the species’ range. Finally, we find that introgression between species is asymmetric, with more from D. recens into D. subquinaria than vice versa. Within each species, admixture is highest in the geographic region where it is rare relative to the other species, suggesting that when hybrids are produced they are of low fitness. In sum, reinforcement within D. subquinaria is effective at maintaining species boundaries, but even when reinforcing selection is strong it may not always result in a pattern of strong reproductive character displacement due to variation in the frequency of hybridization and gene flow from neighbouring populations.