Reconstruction of an enigmatic Pennsylvanian cone reveals a relationship to Sphenophyllales.

PREMISE: We studied the 3D morphology of a small, well-preserved cone from the Pennsylvanian Mazon Creek Lagerstätte to characterize its structure and determine its systematic affinity. Previously tentatively assigned to the enigmatic Tetraphyllostrobus, we show that it differs in key respects from that genus as described. METHODS: We systematically compared the new fossil with relevant Paleozoic cone genera and employed advanced imaging techniques, including scanning electron microscopy, Airyscan confocal super-resolution microscopy, optical microscopy, and X-ray microcomputed tomography to visualize and reconstruct the fossil cone in 3D. RESULTS: The analyses demonstrate unequivocally that the sporophylls of the new Mazon Creek cone are arranged in whorls of six and have characters typical of Sphenophyllales, including epidermal cells with undulatory margins and in situ spores assignable to Columinisporites. The combination of characters, including sporophyll arrangement, anatomy, and spore type, supports the establishment of Hexaphyllostrobus kostorhysii gen. et sp. nov. within Sphenophyllales. Furthermore, we show that Tetraphyllostrobus, although originally described as possessing smooth monolete spores, actually possesses Columinisporites-type spores, indicating that it, too, was most likely a sphenophyll. CONCLUSIONS: The recognition of Hexaphyllostrobus contributes to our knowledge of Pennsylvanian sphenophyll diversity, and in particular increases the number of species with in situ Columinisporites-type spores. Attribution of Hexaphyllostrobus to Sphenophyllales calls into question current interpretations of Tetraphyllostrobus suggesting that future research on better-preserved macrofossil material may demonstrate a sphenophyllalean relationship.

Cavitation fatigue in conifers: a study on eight European species.

After drought-induced embolism and repair, tree xylem may be weakened against future drought events (cavitation fatigue). As there are few data on cavitation fatigue in conifers available, we quantified vulnerability curves (VCs) after embolism/repair cycles on eight European conifer species. We induced 50% and 100% loss of conductivity (LC) with a cavitron, and analyzed VCs. Embolism repair was obtained by vacuum infiltration. All species demonstrated complete embolism repair and a lack of any cavitation fatigue after 50% LC . After 100% LC, European larch (Larix decidua), stone pine (Pinus cembra), Norway spruce (Picea abies), and silver fir (Abies alba) remained unaffected, while mountain pine (Pinus mugo), yew (Taxus baccata), and common juniper (Juniperus communis) exhibited 0.4-0.9 MPa higher vulnerability to embolism. A small cavitation fatigue observed in Scots pine (Pinus sylvestris) was probably biased by incomplete embolism repair, as indicated by a correlation of vulnerability shifts and conductivity restoration. Our data demonstrate that cavitation fatigue in conifers is species-specific and depends on the intensity of preceding LC. The lack of fatigue effects after moderate LC, and relevant effects in only three species after high LC, indicate that conifers are relatively resistant against cavitation fatigue. This is remarkable considering the complex and delicate conifer pit architecture and may be important considering climate change projections.

Detailed seed cone morpho-anatomy of the Prumnopityoid clade: an insight into the origin and evolution of Podocarpaceae seed cones.

BACKGROUND AND AIMS: Seed cone traits are significant for understanding the evolutionary history of conifers. Podocarpaceae has fleshy cones with a distinct morphology compared with other conifers. However, we have a poor understanding of the seed cone morphology of the Prumnopityoid clade and within Podocarpaceae. This study presents detailed seed cone morpho-anatomy and the evolution of fleshy structures traits in the Prumnopityoid clade. METHODS: We investigated the detailed seed cone morpho-anatomy of selected species from the nine genera using the histological method. The evolution of morpho-anatomical traits was assessed using ancestral state reconstruction methods. KEY RESULTS: The Prumnopityoid clade has evolved fleshy seed cones using different functional structures (e.g. aril, epimatium, bracts or receptaculum) and fleshiness is an ancestral trait in the clade. An epimatium is present in all genera except Phyllocladus, but with different structural morphologies (e.g. a fleshy asymmetrical cup-like epimatium or an epimatium that is fused with the integument, forming a fleshy sarcotesta-like seed coat). In all species with fleshy sarcotesta-like seed coats, the endotesta is hard and woody, forming a sclerotesta-like structure and the epimatium and exotesta are fused, forming a fleshy sarcotesta-like structure. CONCLUSIONS: This study highlights that the Prumnopityoid clade has an amazing diversity of structures and complex evolutionary patterns. Fleshiness is an ancestral trait of the clade and has been achieved via diverse evolutionary pathways and structures. This clade has four distinct seed cone types, i.e. drupe-like, receptaculate, arilloid and dacrydioid cones, based on morpho-anatomical structures and traits. The macrofossil record also demonstrates the presence of several structures and traits.

Evolution of the coniferous seed scale.

BACKGROUND: The Florin model is the commonly accepted theory of coniferous seed scale evolution. It describes the derivation of extant seed scale morphology from the morphology of fossil conifers via the reduction of complex to simple axillary structures. In this framework the seed scale is composed of a reduced lateral shoot with fertile and sterile appendages which are interpreted as leaf homologues. SCOPE: The Florin model has three crucial problems that we address here: (1) the original derivation series does not take the ontogeny of extant conifers into account, (2) it cannot explain the morphology of all extant conifers and (3) Taxaceae were originally excluded. Examination of seed cones of extant conifers shows that ovules occur in three different positions in the cone: in an axillary position, replacing a leaf or terminating the cone axis. By interpreting the fertile appendage or seed-bearing structure as a leaf, not all positions are possible. The exclusion of Taxaceae from conifers is in stark contrast to recent molecular phylogenetic studies, which include Taxaceae in conifers as sister to Cupressaceae. Therefore, the Florin model does not offer an adequate explanation for taxaceous morphology. CONCLUSION: We conclude that the seed-bearing structure of conifers cannot be interpreted as homologous to a leaf. In the interpretation we present here, the seed-bearing structure is the modified funiculus of the ovule, multiples of which laterally fuse to form the seed scale. The seed scales of all extant conifers can be derived from a Cunninghamia-like morphology via fusion and reduction of individual funiculi.

Community Assembly and Climate Mismatch in Late Quaternary Eastern North American Pollen Assemblages.

Plant community response to climate change ranges from synchronous tracking to strong mismatch. Explaining this variation in climate change response is critical for accurate global change modeling. Here we quantify how closely assemblages track changes in climate (match/mismatch) and how broadly climate niches are spread within assemblages (narrow/broad ecological tolerance, or "filtering") using data for the past 21,000 years for 531 eastern North American fossil pollen assemblages. Although climate matching has been strong over the last 21 millennia, mismatch increased in 30% of assemblages during the rapid climate shifts between 14.5 and 10 ka. Assemblage matching rebounded toward the present day in 10%-20% of assemblages. Climate-assemblage mismatch was greater in tree-dominated and high-latitude assemblages, consistent with persisting populations, slower dispersal rates, and glacial retreat. In contrast, climate matching was greater for assemblages comprising taxa with higher median seed mass. More than half of the assemblages were climatically filtered at any given time, with peak filtering occurring at 8.5 ka for nearly 80% of assemblages. Thus, vegetation assemblages have highly variable rates of climate mismatch and filtering over millennial scales. These climate responses can be partially predicted by species' traits and life histories. These findings help constrain predictions for plant community response to contemporary climate change.

Assessing the effects of water quality on leaf morphoanatomy, ultrastructure and photosynthetic pigment content of Salvinia auriculata Aubl. (Salviniaceae).

The objective of this study is to evaluate the effects of physical-chemical and biological variables of the water of the Capibaribe River (state of Pernambuco, Brazil) on leaf anatomy, including ultrastructure and photosynthetic pigment of Salvinia auriculata. Specimens of S. auriculata collected in the Gurjaú River, an area with a low pollution degree, were acclimatized in Hoagland's solution and then subjected to three water samples of the Capibaribe River with different levels of pollution. Twenty-one physical-chemical and biological variables were analyzed according to the Standard Methods for the Examination of Water and Wastewater. The results showed that the samples of the Capibaribe River presented nine parameters that did not comply with the current Brazilian legislation. After 15 days of bioassay, S. auriculata presented variations in mesophyll and cuticle thickness, changes in trichome morphology and accumulation of phenolic compounds. No significant differences were observed for photosynthetic pigment content and leaf length of S. auriculata. Multivariate analyses (PCA and Cluster) showed that the point in the Capibaribe River with the highest number of variables that do not comply with the current legislation was responsible for major structural and chemical changes observed in S. auriculata.

Functional diversity and convergence in the evolution of plant reproductive structures.

Background and Aims: Structures that simultaneously perform many functional roles are likely to show a variety of morphological solutions to these demands, and thus probably exhibit high morphological disparity. In contrast, specialization for a few simple functions should result in a more limited suite of morphologies. We explore this idea using lycopsid reproductive structures, which, throughout their history, have performed a limited set of functional roles compared with the reproductive structures of other plant groups such as seed plants. Methods: We scored living and fossil lycopsid taxa for 18 discrete character measurements and several continuous traits, including sporangium size, supporting axis diameter, and strobilus length and width. We used the discrete characters to construct a multivariate morphospace for lycopsid reproductive morphology through time, and the continuous characters to test whether fossil and extant lycopsids show similar patterns of tissue allocation within reproductive structures. Results: Lycopsids occupy similar areas of reproductive morphospace and show similar patterns of tissue allocation over most of their history, alternating between diffuse fertile zones with leaf-like sporophylls and compact strobili with specialized sporophylls that allow sporangia to be closely packed while also protected during their development. Growth habit also plays an important role in lycopsid reproductive evolution, broadly influencing the size and shape of reproductive structures. Conclusions: Lycopsid reproductive structures are primarily specialized for densely packaging sporangia, and are consistent with the idea that performing limited functional roles is associated with reduced morphological disparity. Morphologies similar to lycopsid strobili are also found in other groups with simple, wind-dispersed propagules, suggesting that the same processes occur across plant lineages.

Permineralized conifer-like leaves from the Jurassic of Patagonia (Argentina) and its paleoenvironmental implications.

Anatomically preserved conifer-like leaves from the Middle Jurassic La Matilde Formation at the Barda Blanca locality in the Gran Bajo de San Julián area, southern Patagonia are described here. Leaves are assigned to conifers based on the following foliar features: thick-walled epidermal cells, a sclerenchymatic hypodermis, resin canals and transfusion tracheids associated with the vascular bundle. General mesophyll anatomy and inferred foliar morphology suggest a similarity to large, broad, linear-lanceolate, multi-veined conifer-like leaves. The general foliar habit indicates an affinity with the large, multi-veined leaves of the Araucariaceae; especially with those exhibited by the species of the Araucaria sections, Araucaria and Bunya. Anatomically, the permineralized leaves exhibit xeromorphic foliar features, including thick-walled epidermal cells, an isobilateral mesophyll with well-developed palisade cells and mechanical tissue. The general leaf anatomy shown by the Patagonian specimens along with sedimentological data may suggest that during the deposition of the La Matilde Formation at the Barda Blanca locality, the parent plant was well adapted to the environmental conditions, which probably consisted of a high light intensity with an adequate quantity of water in the soil, which increased the maximum leaf conductance of CO2.

Macrofossil evidence unveiling evolution of male cones in Ephedraceae (Gnetidae).

BACKGROUND: Male cones of modern Ephedraceae are compound and compact. No fossil evidence has so far been found to support an origin of the compact compound male cone from a hypothetical loosely-arranged shoot system. RESULTS: Here we describe a new macrofossil taxon, Eamesia chinensis Yang, Lin, Ferguson et Wang, gen. et sp. nov., from the Early Cretaceous of western Liaoning, northeastern China. It was an ephedroid shrub bearing male spikes terminal to twigs, but differs from modern Ephedraceae by its loosely-arranged male cones, the axillary male shoot consisting of an elongated synangiophore on which leaf-like foliar organs were inserted, and four sessile synangia terminal to the apex. CONCLUSIONS: The morphology of this fossil suggests that the modern compact male cone of Ephedra was indeed derived from a once loosely-arranged shoot system, and the male reproductive unit originated from a once elongated axillary male shoot. This new fossil species thus provides a transitional link from the hypothetical ancestral shoot system to the modern compact morphology. Changes of habitat from closed humid forests to open dry deserts and shifts of the pollination syndrome may have acted as the driving forces behind this morphological evolution.

Why are the seed cones of conifers so diverse at pollination?

Background and Aims: Form and function relationships in plant reproductive structures have long fascinated biologists. Although the intricate associations between specific pollinators and reproductive morphology have been widely explored among animal-pollinated plants, the evolutionary processes underlying the diverse morphologies of wind-pollinated plants remain less well understood. Here we study how this diversity may have arisen by focusing on two conifer species in the pine family that have divergent reproductive cone morphologies at pollination. Methods: Standard histology methods, artificial wind pollination assays and phylogenetic analyses were used in this study. Key Results: A detailed study of cone ontogeny in these species reveals that variation in the rate at which their cone scales mature means that pollination occurs at different stages in their development, and thus in association with different specific morphologies. Pollination experiments nevertheless indicate that both species effectively capture pollen. Conclusions: In wind-pollinated plants, morphological diversity may result from simple variation in development among lineages rather than selective pressures for any major differences in function or performance. This work also illustrates the broader importance of developmental context in understanding plant form and function relationships; because plant reproductive structures perform many different functions over their lifetime, subtle differences in development may dramatically alter the specific morphologies that they use to meet these demands.

Araucaria lefipanensis (Araucariaceae), a new species with dimorphic leaves from the Late Cretaceous of Patagonia, Argentina.

PREMISE OF THE STUDY: We describe a new araucarian species, Araucaria lefipanensis, from the Late Cretaceous flora of the Lefipán Formation, in Patagonia (Argentina) based on reproductive and vegetative remains, with a combination of characters that suggest mosaic evolution in the Araucaria lineage. METHODS: The studied fossils were found at the Cañadón del Loro locality. Specimens were separated into two leaf morphotypes, and their morphological differences were tested with MANOVA. KEY RESULTS: The new species Araucaria lefipanensis is erected based on the association of dimorphic leaves with cuticle remains and isolated cone scale complexes. The reproductive morphology is characteristic of the extant section Eutacta, whereas the vegetative organs resemble those of the sections Intermedia, Bunya, and Araucaria (the broad-leaved clade). CONCLUSIONS: The leaf dimorphism of A. lefipanensis is similar to that of extant A. bidwillii, where dimorphism is considered to be related to seasonal growth. The leaf dimorphism in A. lefipanensis is consistent with the paleoclimatic and paleoenvironmental reconstructions previously suggested for the Lefipán Formation, which is thought to have been a seasonal subtropical forest. The new species shows evidence of mosaic evolution, with cone scale complexes morphologically similar to section Eutacta and leaves similar to the sections of the broad-leaved clade, constituting a possible transitional form between these two well-defined lineages. More complete plant concepts, especially those including both reproductive and vegetative remains are necessary to understand the evolution of ancient plant lineages. This work contributes to this aim by documenting a new species that may add to the understanding of the early evolution of the sections of Araucaria.

An overview of extant conifer evolution from the perspective of the fossil record.

PREMISE OF THE STUDY: Conifers are an important living seed plant lineage with an extensive fossil record spanning more than 300 million years. The group therefore provides an excellent opportunity to explore congruence and conflict between dated molecular phylogenies and the fossil record. METHODS: We surveyed the current state of knowledge in conifer phylogenetics to present a new time-calibrated molecular tree that samples ~90% of extant species diversity. We compared phylogenetic relationships and estimated divergence ages in this new phylogeny with the paleobotanical record, focusing on clades that are species-rich and well known from fossils. KEY RESULTS: Molecular topologies and estimated divergence ages largely agree with the fossil record in Cupressaceae, conflict with it in Araucariaceae, and are ambiguous in Pinaceae and Podocarpaceae. Molecular phylogenies provide insights into some fundamental questions in conifer evolution, such as the origin of their seed cones, but using them to reconstruct the evolutionary history of specific traits can be challenging. CONCLUSIONS: Molecular phylogenies are useful for answering deep questions in conifer evolution if they depend on understanding relationships among extant lineages. Because of extinction, however, molecular datasets poorly sample diversity from periods much earlier than the Late Cretaceous. This fundamentally limits their utility for understanding deep patterns of character evolution and resolving the overall pattern of conifer phylogeny.

Variation in seed size is structured by dispersal syndrome and cone morphology in conifers and other nonflowering seed plants.

Seed size varies tremendously in plants and its evolution is influenced by multiple ecological and biological factors that are difficult to disentangle. In this study, we focus on understanding the role of seed dispersal by animals in the evolution of seed size in conifers, the most diverse extant nonflowering seed plant group. Relationships among seed size, dispersal syndrome, climate and cone morphology were analyzed across conifers using quantitative models of character evolution and phylogenetic regression techniques. Dispersal syndrome is a more consistent predictor of seed size within major extant conifer clades than climate. Seeds are generally larger in animal-dispersed than wind-dispersed species, and particular cone morphologies are consistently associated with specific ranges in seed size. Seed size and cone morphology evolve in a correlated manner in many animal-dispersed conifers, following a trade-off that minimizes the total size of the dispersal unit. These relationships are also present in other nonflowering seed plant groups, and have been important in the evolution of seeds and cones at least over the Cenozoic and perhaps over much of the later Mesozoic.

The fossil flip-leaves (Retrophyllum, Podocarpaceae) of southern South America.

PREMISE OF THE STUDY: The flip-leaved podocarp Retrophyllum has a disjunct extant distribution in South American and Australasian tropical rainforests and a Gondwanic fossil record since the Eocene. Evolutionary, biogeographic, and paleoecological insights from previously described fossils are limited because they preserve little foliar variation and no reproductive structures. METHODS: We investigated new Retrophyllum material from the terminal Cretaceous Lefipán, the early Eocene Laguna del Hunco, and the early/middle Eocene Río Pichileufú floras of Patagonian Argentina. We also reviewed type material of historical Eocene fossils from southern Chile. KEY RESULTS: Cretaceous Retrophyllum superstes sp. nov. is described from a leafy twig, while Eocene R. spiralifolium sp. nov. includes several foliage forms and a peduncle with 13 pollen cones. Both species preserve extensive damage from sap-feeding insects associated with foliar transfusion tissue. The Eocene species exhibits a suite of characters linking it to both Neotropical and West Pacific Retrophyllum, along with several novel features. Retrophyllum araucoensis (Berry) comb. nov. stabilizes the nomenclature for the Chilean fossils. CONCLUSIONS: Retrophyllum is considerably older than previously thought and is a survivor of the end-Cretaceous extinction. Much of the characteristic foliar variation and pollen-cone morphology of the genus evolved by the early Eocene. The mixed biogeographic signal of R. spiralifolium supports vicariance and represents a rare Neotropical connection for terminal-Gondwanan Patagonia, which is predominantly linked to extant Australasian floras due to South American extinctions. The leaf morphology of the fossils suggests significant drought vulnerability as in living Retrophyllum, indicating humid paleoenvironments.

A new Cheirolepidiaceae (Coniferales) from the Early Jurassic of Patagonia (Argentina): Reconciling the records of impression and permineralized fossils.

PREMISE OF THE STUDY: Plants preserved in different fossil modes provide complementary data concerning the paleobiology and evolutionary relationships among plant groups. New material from the Early Jurassic of Patagonia shows the importance of combining these sources of information, as we describe the first compression/impression fossils of Pararaucaria, a genus of the extinct conifer family Cheirolepidiaceae previously known from permineralized fossils. These fossils extend the temporal range of this genus and may allow its wider recognition in the fossil record. METHODS: We studied fossil plants from the Early Jurassic (Pleinsbachian-Toarcian) locality of Taquetrén in Patagonia, Argentina using standard paleobotanical preparation and description techniques. KEY RESULTS: Pararaucaria taquetrensis consists of isolated ovuliferous scales and small seed cones with helically arranged bract-scale complexes attached to scale-leaf foliage. Bract-scale complexes consist of separated bracts and ovuliferous scales with two seeds and three broad distal lobes. CONCLUSIONS: Pararaucaria taquetrensis represents the oldest known Cheirolepidiaceae seed cones from the Southern Hemisphere, and this material highlights the importance of compression and impression fossils in understanding the distribution of fossil taxa. This material also suggests that Cheirolepidiaceae cone scales can be easily confused with those of another common conifer family, the Araucariaceae, which has important implications for accurately understanding Mesozoic conifer diversity and paleoecology.

The economy of reproduction in dimorphic ferns.

BACKGROUND AND AIMS: Organisms often balance among reproduction, growth and survival. When these processes are in competition, selection may act to drive functional dimorphism. Unlike seed plants, ferns use their foliar surfaces for reproduction and carbon fixation. Across species, ferns exhibit a gradient of fertile-sterile dimorphy: from the production of highly reduced fertile fronds (holodimorphic) to no reduction (monomorphic) in laminar area between fronds. Here the physiological impacts of fertile-sterile dimorphy were investigated through a series of observational and experimental field manipulations. METHODS: Temporal shifts in photosynthesis, respiration and percent nitrogen (%N) were examined to evaluate changes in physiological behaviour over the growing season of two species of fern of similar ecological niche, yet of different degrees of fertile-sterile frond dimorphism: Osmundastrum cinnamomeum (holodimorphic) and Osmunda regalis (hemidimorphic). These data are combined with experimental fertile and sterile frond removal to evaluate relative costs of reproduction in both species. Finally, labelled δ13C gas was used to follow carbon allocation across the growing season. KEY RESULTS: The data demonstrate that reproductive structures in the holodimorphic O. cinnamomeum come at more significant carbon and nitrogen costs relative to those in the hemidimorphic O. regalis Excision experiments demonstrate that investment in fertile fronds strongly impacted future allocation to reproduction in the holodimorphic species but had a lesser effect on the hemidimorphic species. The labelling experiments showed that fixed carbon is translocated to the rhizomes only, but at different times in the two species. Investment in underground resources probably allows these plants to manage the costs of reproduction associated with increased dimorphy. CONCLUSIONS: Fertile-sterile dimorphy has evolved multiple times in ferns in spite of the apparent physiological costs associated with a reduction in photosynthetically active tissues. These apparent costs may be offset by an increase in potential spore dispersal distance and/or increased spore production. The phenomenon may further influence species ecology as dimorphic taxa often occupy resource-rich environments.

Phylogenetic diversification of Early Cretaceous seed plants: The compound seed cone of Doylea tetrahedrasperma.

PREMISE OF THE STUDY: Discovery of cupulate ovules of Doylea tetrahedrasperma within a compact, compound seed cone highlights the rich diversity of fructification morphologies, pollination biologies, postpollination enclosure of seeds, and systematic diversity of Early Cretaceous gymnosperms. METHODS: Specimens were studied using the cellulose acetate peel technique, three-dimensional reconstructions (in AVIZO), and morphological phylogenetic analyses (in TNT). KEY RESULTS: Doylea tetrahedrasperma has bract/fertile short shoot complexes helically arranged within a compact, compound seed cone. Complexes diverge from the axis as a single unit and separate distally into a free bract tip and two sporophylls. Each sporophyll bears a single, abaxial seed, recurved toward the cone axis, that is enveloped after pollinaton by sporophyll tissue, forming a closed cupule. Ovules are pollinated by bisaccate grains captured by micropylar pollination horns. CONCLUSIONS: The unique combination of characters shown by D. tetrahedrasperma includes the presence of cupulate seeds borne in conifer-like compound seed cones, an ovuliferous scale analogue structurally equivalent to the ovulate stalk of Ginkgo biloba, gymnospermous pollination, and nearly complete enclosure of mature seeds. These features characterize the Doyleales ord. nov., clearly distinguish it from the seed fern order Corystospermales, and allow for recognition of another recently described Early Cretaceous seed plant as a second species in genus Doylea. A morphological phylogenetic analysis highlights systematic relationships of the Doyleales ord. nov. and emphasizes the explosive phylogenetic diversification of gymnosperms that was underway at the time when flowering plants may have originated and/or first began to radiate.

Cryptic speciation in allotetraploids: Lessons from the Botrychium matricariifolium complex.

PREMISE OF THE STUDY: Cryptic species are a challenge for botanists and taxonomists. To improve species delineation in the genus Botrychium (Ophioglossaceae), which includes multiple instances of allotetraploid speciation, we examined a cryptic species complex using genetics and morphology. METHODS: We sampled species in the B. matricariifolium complex, concentrating on the Upper Peninsula of Michigan and including multiple proposed morphospecies. We analyzed over 1500 samples using 10 enzyme systems, measured 42 quantitative and qualitative morphological characters for over 650 individuals, and analyzed 145 samples using AFLPs. We tested for diagnostic enzymes in the morphospecies and calculated the correlation between morphological and genetic distances to determine whether putatively distinct morphotypes warrant taxonomic recognition. KEY RESULTS: Allozyme allelic variation corresponded loosely to some morphotypes of B. matricariifolium, but with lower genetic distinction among them than found between B. matricariifolium and B. michiganense. Botrychium michiganense contains unique alleles, indicating a different hybrid origin from that of B. matricariifolium and supporting its status as a genetically distinct species. CONCLUSIONS: We showed that B. acuminatum morphology and genetics are accommodated taxonomically within B. matricariifolium; B. matricariifolium and B. michiganense likely represent hybridization events between related species; and morphotypes within B. matricariifolium likely represent repeated hybridization events between the same two parental species. These hybridizations have resulted in the array of morphotypes observed by field botanists. By helping to identify diagnostic morphological characters, genetic analyses also help us understand and resolve morphological variation observed in the field.