BackgroundThe taxonomy and systematics of Salix subgenus Salix s.l. is difficult. The reliability and evolutionary implications of two important morphological characters (number of stamens, and morphology of bud scales) used in subgeneric classification within Salix remain untested, and a disjunct Old–New World distribution pattern of a main clade of subgenus Salix s.l., revealed by a previous study, lacks a reasonable explanation. To study these questions, we conducted phylogenetic analyses based on 4,688 bp of sequence data from four plastid (rbcL, trnD–T, matK, and atpB–rbcL) and two nuclear markers (ETS and ITS) covering all subgenera of Salix, and all sections of subgenus Salix s.l.ResultsSubgenus Salix came out as para- or polyphyletic in both nrDNA and plastid trees. The plastid phylogeny successfully resolved relationships among the major clades of Salix, but resolution within subgenus Salix s.l. remained low. Nevertheless, three monophyletic groups were identifiable in subgenus Salix s.l.: the ‘main clade’ of subgenus Salix s.l., with New and Old World species being reciprocally monophyletic; the section Triandroides clade; and the subgenus Pleuradenia clade. While nrDNA regions showed higher resolution within subgenus Salix s.l., they failed to resolve subgeneric relationships. Extensive, statistically significant gene-tree incongruence was detected across nrDNA–plastid as well as nrDNA ETS–ITS phylogenies, suggesting reticulate evolution or hybridization within the group. The results were supported by network analyses. Ancestral-state reconstructions indicated that multiple stamens and free bud scales represent the plesiomorphic states within Salix, and that several significant shifts in stamen number and bud scale morphology have occurred.ConclusionsSubgenus Salix s.l. is not monophyletic, and the evolutionary history of the subgenus has involved multiple reticulation events that may mainly be due to hybridization. The delimitation of subgenus Salix s.l. should be redefined by excluding section Triandrae and subgenus Pleuradenia from it. The evolutionary lability of bud-scale morphology and stamen number means that these characters are unreliable bases for classification. The disjunct Old–New World distribution of subgenus Salix s.l. appears to be linked to the profound climatic cooling during the Tertiary, which cut off gene exchange between New and Old World lineages.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-015-0311-7) contains supplementary material, which is available to authorized users.

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Willows (Salix: Salicaceae) form a major ecological component of Holarctic floras and consequently are an obvious target for a DNA-based identification system. We surveyed two to seven plastid genome regions (~3.8 kb; ~3% of the genome) from 71 Salix species across all five subgenera, to assess their performance as DNA barcode markers. Although Salix has a relatively high level of interspecific hybridization, this may not sufficiently explain the near complete failure of barcoding that we observed: only one species had a unique barcode. We recovered 39 unique haplotypes, from more than 500 specimens, that could be partitioned into six major haplotype groups. A unique variant of group I (haplotype 1*) was shared by 53 species in three of five Salix subgenera. This unusual pattern of haplotype sharing across infrageneric taxa is suggestive of either a massive nonrandom coalescence failure (incomplete lineage sorting), or of repeated plastid capture events, possibly including a historical selective sweep of haplotype 1* across taxonomic sections. The former is unlikely as molecular dating indicates that haplotype 1* originated recently and is nested in the oldest major haplotype group in the genus. Further, we detected significant non-neutrality in the frequency spectrum of mutations in group I, but not outside group I, and demonstrated a striking absence of geographical (isolation by distance) effects in the haplotype distributions of this group. The most likely explanation for the patterns we observed involves recent repeated plastid capture events, aided by widespread hybridization and long-range seed dispersal, but primarily propelled by one or more trans-species selective sweeps.

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Salix L. is the largest genus in the family Salicaceae (450 species). Several classifications have been published, but taxonomic subdivision has been under continuous revision. Our goal is to establish the phylogenetic structure of the genus using molecular data on all American willows, using three DNA markers. This complete phylogeny of American willows allows us to propose a biogeographic framework for the evolution of the genus. Material was obtained for the 122 native and introduced willow species of America. Sequences were obtained from the ITS (ribosomal nuclear DNA) and two plastid regions, matK and rbcL. Phylogenetic analyses (parsimony, maximum likelihood, Bayesian inference) were performed on the data. Geographic distribution was mapped onto the tree. The species tree provides strong support for a division of the genus into two subgenera, Salix and Vetrix. Subgenus Salix comprises temperate species from the Americas and Asia, and their disjunction may result from Tertiary events. Subgenus Vetrix is composed of boreo-arctic species of the Northern Hemisphere and their radiation may coincide with the Quaternary glaciations. Sixteen species have ambiguous positions; genetic diversity is lower in subg. Vetrix. A molecular phylogeny of all species of American willows has been inferred. It needs to be tested and further resolved using other molecular data. Nonetheless, the genus clearly has two clades that have distinct biogeographic patterns.

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as soon as possible it is being issued in fascicles that may be held in a looseleaf binder. This mode of publication has the advantage of permitting us to update maps and to add species as more information becomes known. We hope that the atlas will help in planning the re-exploration of known rare plant localities and that it will help in the recognition of unique areas that are characterized by concentrations of rare plants. The mapping of specimens by date of collection, furthermore, gives some idea of the past range of rare species and can help to detect declining populations. Our present information base only enables us to recognize rarity, but it is important to proceed beyond this point as soon as possible to the recognition of Ontario plants that may be threatened, endangered, or already extirpated.The information on which we are basing the atlas is as complete and up-to-date as possible. As plant collectors continue to find new localities for species, however, it will be necessary to make changes in the range and status given for some species. We hope that collectors will call significant new records to our attention. A. Definitions.In defining the following terms we have generally followed the usage recommended by the Committee on the Status of Endangered Wildlife in Canada.The words species and plant are used to refer to Atlas of the Rare Vascular Plants of Ontario/Atlas des plantes vasculaires rares de l'Ontario

Experimental crosses between Salix discolor, S. petiolaris, S. bebbiana, and S. lucida resulted in the synthetic hybrids S. discolor × petiolaris and S. bebbiana × petiolaris and their reciprocals. The cross S. bebbiana × discolor and all crosses involving S. lucida were unsuccessful. Field experiments on the mode of pollination of Salix revealed the presence of wind-borne pollen of S. discolor, S. petiolaris, and S. bebbiana, and seed was set by these species in the absence of insect vectors. They are suspected to be both entomophilous and anemophilous. Salix lucida did not set seeds under these conditions and it is hypothesized that it is fundamentally entomophilous. Evidence is presented that suggests that natural hybridization between S. discolor and S. petiolaris may be restricted by differences in flowering time and by pollen competition.