Putative Parental Species Research Articles

Hybridization processes in an introduced subpopulation of an endangered plant: Management strategies to guarantee the conservation of Helosciadium bermejoi (Apiaceae)

Translocations are a tool to restore populations of threatened plant species that are being widely used. However, these techniques are not without risks and require rigorous protocols to carry them out and for their subsequent monitoring. In translocation projects, hybridization with other species is one of the most important risks and may threaten the survival of the species we want to protect. Here we present the case of Helosciadium bermejoi (L. Llorens) Popper and M.F. Watson, a Critically Endangered plant from the island of Menorca (Western Mediterranean). It occurs only in one locality and has been introduced to three new locations during recent years according to its Recovery Plan (2008). Recently, intermediate forms between the endangered species and a native congener, Helosciadium nodiflorum (L.) Koch, have been detected. The aims of this study were (i) to elucidate whether these plants originated through a hybridization event and (ii) to provide information about the current conservation status of the species. Exhaustive monitoring of all subpopulations (number of patches and area of occupancy) was performed from September 2015 to June 2016, and the rDNA ITS region and cpDNA rps16-trnK intergenic spacer region were sequenced for several samples of the putative hybrid and its putative parental species. Overall, the entire population of H. bermejoi greatly increased from 2010 (last census with available data) to 2015 in terms of both number of patches and area of occupancy: from 110 to 277 patches and from 299.2 to 791.3dm2. Molecular analysis confirmed the hybrid origin of Helosciadium×clandestinum Rita, Capó and Cursach and that both H. bermejoi and H. nodiflorum (L.) Koch can act as donors of pollen or ovules. We discuss the advantages and disadvantages of translocations, as well as the great importance of monitoring them for at least the next ten years, both to know if the actions have been successful and to prevent or mitigate risks. We also present this case as an example for the importance of fast decision and coordination between environmental managers.

Evidence for continual hybridization rather than hybrid speciation between Ligularia duciformis and L.paradoxa (Asteraceae).

BackgroundHybrids possess phenotypic traits that are often intermediate between their parental taxa, which commonly serves as evidence of hybridization in morphological analyses. Natural hybridization has been shown to occur frequently in Ligularia (Asteraceae). In a previous study, Ligularia ×maoniushanensis was demonstrated as a natural hybrid species between L. duciformis and L. paradoxa based on morphological and reproductive traits.MethodsWe used three chloroplast (cpDNA) fragments (psbA-trnH, trnL-rpl32 and trnQ-5′rps16), the nuclear ribosomal internal transcribed spacer (nrITS), and co-dominant SSR and dominant ISSR markers to study natural hybridization between L. duciformis and L. paradoxa growing sympatrically in two locations. Parental taxa were inferred using network analyses of cpDNA and nrITS haplotypes. Admixture among individuals was examined using the Bayesian clustering programs STRUCTURE and NewHybrids based on the SSR and ISSR data; and potential introgression in the SSR loci was assessed using the INTROGRESS package.ResultsThe putative parental species were clearly distinguished from other sympatric Ligularia species by nrITS data, and L. ×maoniushanensis individuals were confirmed to be the hybrid offspring of L. duciformis and L. paradoxa. Moreover, introgression was detected among several individuals morphologically identified as L. duciformis or L. paradoxa. Analyses of the cpDNA data revealed primarily unidirectional hybridization between L. duciformis and L. paradoxa, with L. paradoxa as the maternal parent in Mt. Maoniu, whereas bidirectional but asymmetrical hybridization was inferred to occur in Heihai Lake. The STRUCTURE analyses based on the SSR data detected two distinct clusters among the three taxa. The NewHybrids analyses showed that individuals circumscribed as L. ×maoniushanensis were dominated by early- and later-generation and backcrossing hybrids. The NewHybrids results based on the ISSR data were congruent with SSR results. In addition, introgression was detected in some SSR loci, and heterogeneity among loci was found in terms of detected patterns of introgression.ConclusionsOur data provide strong evidence for hybridization and introgression between L. duciformis and L. paradoxa. Ligularia ×maoniushanensis was demonstrated to be of hybrid origin. Since no evident reproductive isolation was found between the two parental species, detected hybrids appear to be part of hybrid swarms resulting from frequent and ongoing gene flow, which might impede the formation of a new hybrid species.

A taxonomic account of Ranunculus section Batrachium (Ranunculaceae)

In a new taxonomic account of Ranunculus section Batrachium 30 species are recognized. We present a key to the species based on morphological characters of diagnostic value. Taxonomic descriptions including information on chromosome counts, geographic range and ecological preferences are provided. For all species synonyms are listed, allowing access to the relevant taxonomic and floristic literature including taxonomic databases worldwide. Information on types and type localities is provided wherever possible. Reported and confirmed hybrids are listed and assigned to their putative parental species. Infraspecific categories such as subspecies and varieties are not recognized. Unresolved taxonomic problems are discussed for each species and for the section as a whole.

The Origin of New Natural hybrid, Goodyera × maximo-velutina (Orchidaceae) from Jeju Island, Korea

We describe a new taxon of Goodyera (Orchidaceae), G. × maximo-velutina from Jeju Island. This taxon is presumed to be a natural hybrid between G. maximowicziana and G. velutina based on morphological characteristics, i.e., leaf margin, leaf venation, hairiness and color of floral bract, pedicel and ovary. We analyzed ITS of nuclear DNA and three noncoding regions of chloroplast DNA (trnH-psbA, trnS-trnG, trnL) for elucidating the hybrid origin of the new species, G. × maximo-velutina, and two putative parental species. The morphological characteristics and illustrations of the species based on the holotype and other specimens from natural populations are provided together with photographs of the habitat.

Typification and taxonomy of Thymus carstiensis (Lamiaceae)

The name Thymus dalmaticus var. carstiensis, basionym of Th. carstiensis, is here lectotypified. This taxon is currently regarded as the hybrid between Th. longicaulis and Th. pulegioides or as a synonym of Th. serpyllum. A morphological comparison among Th. dalmaticus var. carstiensis and its putative parental species are performed. A detailed morphological description and information about habitat, phenology, distribution and taxonomy are provided. Furthermore, Th. carstiensis is here recorded for the first time from Slovenia.

Similar Morphologies but Different Origins: Hybrid Status of Two More Semi-creeping Taxa of Melastoma.

Inferring the origins of hybrid taxa based on morphology alone is difficult because morphologically similar hybrids can arise from hybridization between different populations of the same parental species or be produced by hybridization of different parental species. In this study, we investigated the origins of two semi-creeping taxa in Melastoma, which are morphologically similar to a natural hybrid, M. intermedium, by sequencing a chloroplast intergenic spacer, nuclear ribosomal internal transcribed spacer and two low-copy nuclear genes (tpi and cam) in these taxa and their putative parental species. Our sequence analysis provides compelling evidence for the hybrid status of the two semi-creeping taxa: one originating from hybridization between M. dodecandrum and M. malabathricum, and the other between M. dodecandrum and M. normale. The origins of these hybrids are therefore clearly different from M. intermedium, and morphological similarity for the three hybrids is most likely due to their origins from hybridization between the same creeping species M. dodecandrum and a different erect species in each of the three cases. We also observed low rate of introgression from M. normale to M. dodecandrum, and genetic exchange between them may transfer adaptive traits to M. dodecandrum. Rare occurrence of these two hybrids may be due to small range overlaps between parental species in one case, and different flowering periods between parental species in the other.

Isolation and characterization of microsatellite loci in Sorbus porrigentiformis and cross-amplification in S. aria and S. rupicola (Rosaceae).

Premise of the study:Southwestern Britain is an emblematic hotspot of polyploid diversity of whitebeams (Sorbus aria agg.; Rosaceae) with ca. 30 polyploid endemic species. The tetraploid S. porrigentiformis is postulated as one of the parents of most of these endemics, along with the sexual diploid S. aria s. str. and the tetraploid S. rupicola.Methods and Results:We isolated 16 nuclear microsatellite loci from S. porrigentiformis and characterized them on 45 trees representing the three putative parental species. Eleven loci were polymorphic, and eight of them exhibited species-specific alleles. Allele numbers ranged from one to 11, and observed heterozygosity ranged from 0.40 to 1.00. The intraspecific levels of variation were very low, in agreement with the facultative apomictic reproduction hypothesized for this species.Conclusions:The species-specific alleles will be useful for tracing the origin of the narrowly distributed Sorbus taxa. In addition, the assessment of diversity levels will help design a conservation strategy for the polyploid complex.

The triparental triploid onion Allium × cornutum Clementi ex Visiani, 1842, possesses a sterile S-type of cytoplasm

Triploid onion, Allium × cornutum Clementi ex Visiani, 1842 (2n = 3x = 24), a vegetatively reproduced garden crop, possess a complex triparental genome organization with three putative parental species, A. cepa L., A. pskemense B. Fedtsch., and A. roylei Stearn. Two of its most studied clones are the Croatian ‘Ljutika’ and the Indian ‘Pran’, which are genetically highly similar. Earlier studies have shown that ‘Pran’ possesses some molecular markers in the chloroplast DNA (cpDNA) identical to those of the unique male-sterile (S) cytoplasm, used for onion breeding. To find out whether ‘Ljutika’ also possesses a S-type of cytoplasm, we analyzed several cpDNA and mitochondrial (mtDNA) molecular markers. The PCR amplification and RFLP analysis of the chloroplast genes accD, atpF, petB and the mitochondrial gene cob, as well as the sequence analysis of the chloroplast matK and atpB-rbcL regions showed that ‘Ljutika’ possesses the male-sterile S-type of cytoplasm. The phylogenetic analysis of the matK and atpB-rbcL sequences of A. × cornutum, its parental species and other Allium species of the section Cepa showed that none of the analyzed species had the identical type of cpDNA as A. × cornutum. Results also suggested that A. pskemense can be excluded as a donor of the S-cytoplasm and a female parent, whereas cpDNA of A. roylei, although not identical to S-cytoplasm, possessed many polymorphisms of S-type. Fluorescent in situ hybridization, using fluorescently labelled parental genomic DNAs as probes in combination with fluorescently labelled 5S and 35S rDNAs enabled simultaneous visualization of the three genomes during meiosis and confirmed their homeologus intergenomic pairing.

Viola woosanensis, a recurrent spontaneous hybrid between V. ulleungdoensis and V. chaerophylloides (Violaceae) endemic to Ulleung Island, Korea.

Ulleung Island is an oceanic volcanic island in Korea, which has never been connected to the adjacent continent. Previous studies highlighted Ulleung Island as an excellent system to study the pattern and process of early stages of flowering plant evolutions on oceanic island. The predominant mode of speciation in flowering plants on Ulleung Island appears to be anagenesis. However, the potentially important role of hybrid speciation among incompletely reproductively isolated lineages cannot be ruled out. Viola woosanensis (Violaceae) is of purportedly hybrid origin between V. ulleungdoensis (i.e., formerly recognized as V. selkirkii in Ulleung Island) and V. chaerophylloides, based on morphology. To examine the origin of V. woosanensis, we sampled a total of 80 accessions, including V. woosanensis and its putative parental species and sequenced nrDNA ITS, and four highly variable chloroplast noncoding regions (trnL-trnF, rpl16 intron, atpF-atpH, and psbA-trnH). Representative species of Viola from Korea were also included in the phylogenetic analyses (maximum parsimony, maximum likelihood, and Bayesian inference). Additive polymorphic sites in the nrDNA ITS regions were confirmed by cloning amplicons from representative species. The molecular data strongly supported the hybrid origin of V. woosanensis, and the maternal and paternal parent were determined to be V. ulleungdoensis and V. chaerophylloides, respectively. The presence of two parental ribotypes in V. woosanensis (with the exception in one population) was confirmed by cloning, suggesting V. woosanensis is primarily the F1 generation. No trace of backcrossing and introgression with its parents was detected due to low fertility of hybrid species. We found a multiple and unidirectional hybrid origin of V. woosanensis. Additional studies are required to determine which factors contribute to asymmetric gene flow of Viola species in Ulleung Island.

Molecular Evidence for the Hybrid Origin of Ilex dabieshanensis (Aquifoliaceae).

Ilex, the largest genus of dioecious woody plants, is a good study system to assess the role of hybridization in speciation and evolution. Ilex dabieshanensis, a tree endemic to Dabieshan Mountains region, was initially described as a new species. Based on morphological intermediacy and sympatric distribution with its putative parental species, I. cornuta and I. latifolia, we proposed it as a natural hybrid between them. In this study, we sequenced one chloroplast intergenic spacer (trnH-psbA) and two nuclear genes (gapC and nepGS) in I. dabieshanensis and its putative parental species to test the hybrid origin hypothesis. Our results showed that there were one to two differentially fixed sequence differences between I. cornuta and I. latifolia at the two nuclear genes. Twelve of the 14 individuals of I. dabieshanensis exhibited additivity in chromatograms on these differentially fixed sites at both nuclear genes, and the remaining two exhibited additivity in chromatograms on the fixed site at only the nepGS gene. Except one haplotype of I. cornuta at the nepGS gene, all of the haplotypes of I. cornuta at the two nuclear genes were well separated from those of I. latifolia, and most haplotypes of I. dabieshanensis were shared with those of I. cornuta and I. latifolia. Phylogenetic analysis of these haplotypes was largely consistent with haplotype network analysis. I. cornuta and I. latifolia differed by two nucleotide substitutions in the chloroplast intergenic spacer, and 12 individuals of I. dabieshanensis had the same sequences as I. latifolia, while the remaining two were identical with I. cornuta. The molecular data provide convincing evidence for the hybrid origin of I. dabieshanensis and asymmetrical direction of hybridization. One haplotype of I. cornuta at the nepGS gene was nested with those of I. latifolia, indicating introgression to I. cornuta.

Two new and one unrecorded natural hybrids between <i>Asplenium ruprechtii</i> and related taxa (Aspleniaceae)

Two new and one unrecorded hybrids of the genus Asplenium were newly found in Korea. They were proved to be hybrid based on intermediate morphology between putative parents. The first hybrid is C.S. Lee & K. Lee (Aspleniaceae), nom. nov. (vernacular name: geo-mi-sa-cheol-go-sa-ri). This new hydrid is a cross between the Asian walking fern, A. ruprechtii and A. pekinens in Uiryeong-gil, Mt. Bukansan, Seoul, Korea. The other new one occurs in Mt. Seongsan in Yeoncheon-gun, Gyeonggi-do, Korea, C.S. Lee & K. Lee, nom. nov. (vernacular name: san-kko-ri-go-sa-ri) which might have experienced multiple hybridization events between A. ruprechtii A. trichomanes and A. incisum following the hybrid between A. ruprechtii and A. trichomanes. In addition, one unrecorded hybrid, Kurata & Hutoh (vernacular name: geo-mi-dol-dam-go-sa-ri), reported first from Japan, is found in natural habitats in Do-dong, Daegu-si, and is a hybrid, between the Asian walking fern and A. sarelli. These hybrid taxa grow in sympatric regions with both putative parental species. The diagnostic characters for each taxon and evidence for their hybridization have been suggested. Descriptions, illustrations, and photographs of these hybrid taxa in their habitats are provided from Korea.

Genome evolution in alpine oat-like grasses through homoploid hybridization and polyploidy.

Hybridization and polyploidization can radically impact genome organization from sequence level to chromosome structure. As a result, often in response to environmental change and species isolation, the development of novel traits can arise and will tend to result in the formation of homoploid or polyploid hybrid species. In this study we focus on evidence of hybridization and polyploidization by ascertaining the species parentage of the endemic alpine Helictotrichon parlatorei group. This group comprises five taxa; the diploids H. parlatorei, Helictotrichon setaceum subsp. setaceum and subsp. petzense, their putative hybrid Helictotrichon ×krischae and the hexaploid Helictotrichon sempervirens. For molecular analyses, cloned nuclear Topoisomerase VI genes of H. sempervirens and H. ×krischae were sequenced and compared with sequences of the diploids to estimate the evolutionary history in this group. In addition, detailed chromosome studies were carried out including fluorescence in situ hybridization (FISH) with 5S and 45S ribosomal and satellite DNA probes, and fluorochrome staining with chromomycin and DAPI. Two distinct types of Topoisomerase VI sequences were identified. One of them (SET) occurs in both subspecies of H. setaceum, the other (PAR) in H. parlatorei. Both types were found in H. ×krischae and H. sempervirens Karyotypes of H. parlatorei and H. setaceum could be distinguished by chromosomes with a clearly differentiated banding pattern of ribosomal DNAs. Both patterns occurred in the hybrid H. ×krischae Hexaploid H. sempervirens shares karyotype features with diploid H. parlatorei, but lacks the expected chromosome characteristics of H. setaceum, possibly an example of beginning diploidization after polyploidization. The geographic origin of the putative parental species and their hybrids and the possible biogeographical spread through the Alps are discussed.

Additivity of ISSR Markers in Natural Hybrids of Related Forest Species Bromus benekenii and B. ramosus (Poaceae)

AbstractThe co-occurrence of hybrids and parental species in similar ecological niches poses a question on the role of traits additivity and overdispersion (emergence of new traits) in microevolutionary processes. We analysed genetic polymorphism ofBromus benekenii,B. ramosusand the spontaneous hybridB. benekenii×B. ramosusin sympatric and allopatric parts of the species distribution in Europe, based on non-coding regions of the taxon genomes (ISSR genetic fingerprinting). We tested 68 individuals in 7 populations, including a hybrid population in N France. Altogether 233 polymorphic ISSR bands (loci) were obtained. We found that the parent species were genetically distinct and the hybrids had an additive pattern of ISSR bands found in the putative parental species (NMDS, STRUCTURE); however, there was evidence of introgression towardsB. ramosus(NEWHYBRIDS, UPGMA classifications, Nei's D genetic distance).Bromus benekeniihad 72,B. ramosus21 and the hybrids 9 private bands (genetic overdispersion), probably resulting from the rearranged genomes. Based on its low genetic divergence index DW, the hybrid population seems to be at a young age. We argue that in the face of anthropogenic landscape transformations favouring secondary contacts, the hybrids may competitively replace the parental species in sympatric areas.

Genetic differentiation and hybrid identification using microsatellite markers in closely related wild species.

Identifying the genetic basis of speciation is critical for understanding the evolutionary history of closely related wild species. Recently diverged species facilitate the study of speciation because many genetic and morphological characteristics are still shared by the organisms under study. The Petunia genus grows in South American grasslands and comprises both recently diverged wild species and commercial species. In this work, we analysed two closely related species: Petunia exserta, which has a narrow endemic range and grows exclusively in rocky shelters, and Petunia axillaris, which is widely distributed and comprises three allopatric subspecies. Petunia axillaris ssp. axillaris and P. exserta occur in sympatry, and putative hybrids between them have been identified. Here, we analysed 14 expressed sequence tag-simple sequence repeats (EST-SSRs) in 126 wild individuals and 13 putative morphological hybrids with the goals of identifying differentially encoded alleles to characterize their natural genetic diversity, establishing a genetic profile for each taxon and to verify the presence of hybridization signal. Overall, 143 alleles were identified and all taxa contained private alleles. Four major groups were identified in clustering analyses, which indicated that there are genetic distinctions among the groups. The markers evaluated here will be useful in evolutionary studies involving these species and may help categorize individuals by species, thus enabling the identification of hybrids between both their putative taxa. The individuals with intermediate morphology presented private alleles of their both putative parental species, although they showed a level of genetic mixing that was comparable with some of the individuals with typical P. exserta morphology. The EST-SSR markers scattered throughout the Petunia genome are very efficient tools for characterizing the genetic diversity in wild taxa of this genus and aid in identifying interspecific hybrids based on the presence of private alleles. These properties indicate that these markers will be helpful tools in evolutionary studies.

Natural hybridization and introgression between Ligularia cymbulifera and L. tongolensis (Asteraceae, Senecioneae) in four different locations.

Natural hybridization has been considered to represent an important factor influencing the high diversity of the genus Ligularia Cass. in the Hengduan Mountains, China. Natural hybridization has been confirmed to occur frequently in Ligularia. To date, however, it has been demonstrated only within a single population. In this paper, we present evidence of natural hybridization in Ligularia from four different locations. The internal transcribed spacer (ITS) region of the nuclear ribosomal DNA and three chloroplast intergenic spacers (trnK-rps16, trnL-rpl32 and trnQ-5'rps16) of 149 accessions of putative hybrids and their putative parents (L. cymbulifera and L. tongolensis) were analyzed for evidence of hybridization. The ITS data clearly distinguished two putative parental species and sympatric L. vellerea and supported the hypothesis that those morphological intermediates were products of natural hybridization between L. cymbulifera and L. tongolensis. Moreover, several identified morphological parents were actual introgressed products. Because of hybridization and introgression, chloroplast DNA sequences generated a poorly resolved network. The present results indicate that varying degrees of hybridization and introgression occur differently depending on the habitat context. We conclude that gene flow caused by natural hybridization in Ligularia indeed plays an important role in the species diversity.

Hybridisation and diversification in the adaptive radiation of clownfishes.

BackgroundThe importance of hybridisation during species diversification has long been debated among evolutionary biologists. It is increasingly recognised that hybridisation events occurred during the evolutionary history of numerous species, especially during the early stages of adaptive radiation. We study the effect of hybridisation on diversification in the clownfishes, a clade of coral reef fish that diversified through an adaptive radiation process. While two species of clownfish are likely to have been described from hybrid specimens, the occurrence and effect of hybridisation on the clade diversification is yet unknown.ResultsWe generate sequences of three mitochondrial genes to complete an existing dataset of nuclear sequences and document cytonuclear discordance at a node, which shows a drastic increase of diversification rate. Then, using a tree-based jack-knife method, we identify clownfish species likely stemming from hybridisation events. Finally, we use molecular cloning and identify the putative parental species of four clownfish specimens that display the morphological characteristics of hybrids.ConclusionsOur results show that consistently with the syngameon hypothesis, hybridisation events are linked with a burst of diversification in the clownfishes. Moreover, several recently diverged clownfish lineages likely originated through hybridisation, which indicates that diversification, catalysed by hybridisation events, may still be happening.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-014-0245-5) contains supplementary material, which is available to authorized users.

Nuclear and Cytoplasmic DNA Sequence Data Further Illuminate the Genetic Composition of Leyland Cypresses

Leyland cypress [×Hesperotropsis leylandii (A.B. Jacks. & Dallim.) Garland & G. Moore, Cupressaceae] is a well-known horticultural evergreen conifer in the United Kingdom, United States, Australia, New Zealand, and other countries. As demonstrated by previous studies, this taxon is a hybrid between alaska (nootka) cypress [Callitropsis nootkatensis (D. Don) Oerst. ex D.P. Little] and monterey cypress [Hesperocyparis macrocarpa (Hartw. ex Gordon) Bartel]. However, the genetic background of leyland cypress cultivars is unclear. Are they F1 or F2 hybrids or backcrosses? In this study, six individuals that represent major leyland cypress cultivars and two individuals each of its two putative parental species were collected, and three nuclear DNA regions (internal transcribed spacer, leafy and needly), three mitochondrial (mt) DNA regions (coxI, atpA, and rps3), and two chloroplast (cp) DNA regions (matK and rbcL) were sequenced and analyzed. Sequencing results of nuclear DNA regions revealed that leyland cypress cultivars consist of putative F1 and F2 hybrids as well as backcrosses. Analysis of the cp and mt DNA from six cultivars of leyland cypress revealed that their cytoplasmic (cp and mt) genomes came from alaska cypress. Our findings will provide important instructions and background knowledge on the management of these major leyland cypress cultivars as well as future studies. Meanwhile, alaska cypress and monterey cypress may have diverged with each other ≈46 million years ago. The fact that they can produce fertile hybrids indicates that hybridization events may have played an important role in the evolutionary history of the cypress family (Cupressaceae).

Extensive hybridization and introgression between Melastoma candidum and M. sanguineum.

Natural hybridization can lead to various evolutionary outcomes in plants, including hybrid speciation and interspecific gene transfer. It can also cause taxonomic problems, especially in plant genera containing multiple species. In this study, the hybrid status of Melastoma affine, the most widespread taxon in this genus, and introgression between its putative parental species, M. candidum and M. sanguineum, were assessed on two sites, Hainan and Guangdong, using 13 SSR markers and sequences of a chloroplast intergenic spacer. Bayesian-based STRUCTURE analysis detected two most likely distinct clusters for the three taxa, and 76.0% and 73.9% of the morphologically identified individuals of M. candidum and M. sanguineum were correctly assigned, respectively. 74.5% of the M. affine individuals had a membership coefficient to either parental species between 0.1 and 0.9, suggesting admixture between M. candidum and M. sanguineum. Furthermore, NewHybrids analysis suggested that most individuals of M. affine were F2 hybrids or backcross hybrids to M. candidum, and that there was extensive introgression between M. candidum and M. sanguineum. These SSR data thus provides convincing evidence for hybrid origin of M. affine and extensive introgression between M. candidum and M. sanguineum. Chloroplast DNA results were consistent with this conclusion. Much higher hybrid frequency on the more disturbed Guangdong site suggests that human disturbance might offer suitable habitats for the survival of hybrids, a hypothesis that is in need of further testing.

A new hybrid from Taiwan, Elatostema ×hybrida (Urticaceae), is the first confirmed natural hybrid for Urticaceae

Explosive pollen dispersal is common in Urticaceae and they are thought to be wind-pollinated. Despite a lack of obvious mechanism for preventing cross-species pollination, putative hybrid species in Urticaceae are rarely documented. Here we described the first natural hybrid in Urticaceae Elatostema ×hybrida from Taiwan. Morphological characters in E. ×hybrida are intermediate between putative parental species: E. lineolatum var. majus and E. platyphylloides. Six hybrid populations of E. ×hybrida were found in Taiwan that exhibited largely overlapping distribution patterns with its putative parents. Phylogenetic analysis of chloroplast DNA showed that the hybrid species is more closely related to E. lineolatum var. majus suggesting that the latter is the maternal parent and that hybridization is unidirectional. The chromosome number of E. ×hybrida remains the same as its putative parents (2n = 26). We speculate that the examined hybrids are natural first-generation results of independent hybridization events. Based on the morphology, spatial distribution, DNA sequence data, pollen viability and cytological observations, we hypothesize that E. ×hybrida is derived from natural hybridization events between E. lineolatum var. majus (♀) and E. platyphylloides (♂).

Triparental origin of triploid onion, Allium × cornutum (Clementi ex Visiani, 1842), as evidenced by molecular, phylogenetic and cytogenetic analyses

BackgroundReconstruction of the parental origins of cultivated plants from wild relatives, especially after long periods of domestication, is not a trivial task. However, recent advances in molecular phylogenetics, among other approaches, have proved to be very informative in analyses of the origin and evolution of polyploid genomes. An established minor garden crop, triploid onion Allium × cornutum (Clementi ex Visiani, 1842) (2n = 3x = 24), is widespread in southeastern Asia and Europe. Our previous cytogenetic analyses confirmed its highly heterozygous karyotype and indicated its possible complex triparental genome origin. Allium cepa L. and Allium roylei Stearn were suggested as two putative parental species of A. × cornutum, whereas the third parental species remained hitherto unknown.ResultsHere we report the phylogenetic analyses of the internal transcribed spacers ITS1-5.8S-ITS2 of 35S rDNA and the non-transcribed spacer (NTS) region of 5S rDNA of A. × cornutum and its relatives of the section Cepa. Both ITS and NTS sequence data revealed intra-individual variation in triploid onion, and these data clustered into the three main clades, each with high sequence homology to one of three other species of section Cepa: A. cepa, A. roylei, and unexpectedly, the wild Asian species Allium pskemense B. Fedtsh. Allium pskemense is therefore inferred to be the third, so far unknown, putative parental species of triploid onion Allium × cornutum. The 35S and 5S rRNA genes were found to be localised on somatic chromosomes of A. × cornutum and its putative parental species by double fluorescent in situ hybridisation (FISH). The localisation of 35S and 5S rDNA in A. × cornutum chromosomes corresponded to their respective positions in the three putative parental species, A. cepa, A. pskemense, and A. roylei. GISH (genomic in situ hybridisation) using DNA of the three putative parental diploids corroborated the results of the phylogenetic study.ConclusionsThe combined molecular, phylogenetic and cytogenetic data obtained in this study provided evidence for a unique triparental origin of triploid onion A. × cornutum with three putative parental species, A. cepa, A. pskemense, and A. roylei.