Hymenasplenium volubile: documentation of its gametophytes and the first record of a hemiepiphyte in the Aspleniaceae.

BACKGROUND AND AIMS: Through careful field examination of the growth habit of the gametophytes and sporophytes of Hymenasplenium volubile across an ontogenetic series, we aim to understand better the evolution of epiphytism in this poorly understood group of ferns. METHODS: We made field observations of H. volubile sporophytes and gametophytes, and brought specimens back to the lab for microscopic analysis. In the field, sporophytes at each ontogenetic stage were photographed to document the species' growth habit. We used an existing phylogeny to optimize growth form of New World Hymenasplenium. KEY RESULTS: Young sporophytes were at first fully epiphytic and produced one or two long feeding roots that extend to the soil where they branch profusely. The feeding roots remain in contact with the soil throughout the life of the plant. Thus, H. volubile is a hemiepiphyte. While immature, gametophytes are appressed to the tree trunk, but, as their gametangia mature, their lower margin lifts upward, imparting a shelf-like appearance to the thallus. The thallus attaches to the substrate by branched rhizoids produced along the margin of the thallus in contact with the substrate. CONCLUSIONS: Hemiepiphytes are a key link in the evolution of epiphytic ferns and may act as a bridge between the forest floor and the canopy. Our finding is the first report of hemiepiphytism in Aspleniaceae, a large lineage with many epiphytic and terrestrial taxa. This work serves as an important model to understand the evolution of epiphytism in this group specifically and in ferns in general. The majority of our understanding of fern gametophyte biology is derived from laboratory studies. Our efforts represent a fundamental contribution to understanding fern gametophyte ecology in a field setting.

An exquisitely preserved filmy fern (Hymenophyllaceae) from the Early Cretaceous of Mongolia.

PREMISE OF THE STUDY: Hymenophyllaceae ("filmy ferns") are a widely distributed group of predominantly tropical, epiphytic ferns that also include some temperate and terrestrial species. Hymenophyllaceae are one of the earliest-diverging lineages within leptosporangiate ferns, but their fossil record is sparse, most likely because of their low fossilization potential and commonly poor preservation of their delicate, membranaceous fronds. A new species of unequivocal fossil Hymenophyllaceae, Hymenophyllum iwatsukii sp. nov., is described from the Early Cretaceous of Mongolia based on abundant and exceptionally well-preserved material. METHODS: Bulk lignite samples collected from Tevshiin Govi and Tugrug localities in Mongolia, were disaggregated in water, cleaned with hydrochloric and hydrofluoric acids, washed, and dried in air. Fossils were examined and compared to material of extant Hymenophyllaceae using LM and SEM. KEY RESULTS: The fossil fern specimens are assigned to the Hymenophyllaceae based on their membranaceous laminae with marginal sori that have sessile to short-stalked sporangia with oblique, complete annuli, and trilete, tetrahedral-globose spores. Within the family, the fossil material is assigned to the extant genus Hymenophyllum on the basis of bivalvate indusia and short, included receptacles. CONCLUSIONS: Hymenophyllum iwatsukii was likely an epiphyte based on the sedimentary environment in which the fossils are preserved, the associated fossil flora, and the growth habit of extant species of Hymenophyllum. The new fossil species underlines the extent to which morphological characters in Hymenophyllum have been conserved despite significant tectonic, climatic, ecological, and floristic changes since the Early Cretaceous.

Challenging the paradigms of leaf evolution: Class III HD-Zips in ferns and lycophytes.

Despite the extraordinary significance leaves have for life on Earth, their origin and development remain vigorously debated. More than a century of paleobotanical, morphological, and phylogenetic research has still not resolved fundamental questions about leaves. Developmental genetic data are sparse in ferns, and comparative studies of lycophytes and seed plants have reached opposing conclusions on the conservation of a leaf developmental program. We performed phylogenetic and expression analyses of a leaf developmental regulator (Class III HD-Zip genes; C3HDZs) spanning lycophytes and ferns. We show that a duplication and neofunctionalization of C3HDZs probably occurred in the ancestor of euphyllophytes, and that there is a common leaf developmental mechanism conserved between ferns and seed plants. We show C3HDZ expression in lycophyte and fern sporangia and show that C3HDZs have conserved expression patterns during initiation of lateral primordia (leaves or sporangia). This expression is maintained throughout sporangium development in lycophytes and ferns and indicates an ancestral role of C3HDZs in sporangium development. We hypothesize that there is a deep homology of all leaves and that a sporangium-specific developmental program was coopted independently for the development of lycophyte and euphyllophyte leaves. This provides molecular genetic support for a paradigm shift in theories of lycophyte leaf evolution.

The first fossil of a bolbitidoid fern belongs to the early-divergent lineages of Elaphoglossum (Dryopteridaceae).

UNLABELLED: • PREMISE OF THE STUDY: Closing gaps in the fossil record and elucidating phylogenetic relationships of mostly incomplete fossils are major challenges in the reconstruction of the diversification of fern lineages through time. The cosmopolitan family Dryopteridaceae represents one of the most species-rich families of leptosporangiate ferns, yet its fossil record is sparse and poorly understood. Here, we describe a fern inclusion in Miocene Dominican amber and investigate its relationships to extant Dryopteridaceae.• METHODS: The morphology of the fossil was compared with descriptions of extant ferns, resulting in it being tentatively assigned to the bolbitidoid fern genus Elaphoglossum. This assignment was confirmed by reconstructing the evolution of the morphological characters preserved in the inclusion on a molecular phylogeny of 158 extant bolbitidoid ferns. To assess the morphology-based assignment of the fossil to Elaphoglossum, we examined DNA-calibrated divergence time estimates against the age of the amber deposits from which it came.• KEY RESULTS: The fossil belongs to Elaphoglossum and is the first of a bolbitidoid fern. Its assignment to a particular section of Elaphoglossum could not be determined; however, sects. Lepidoglossa, Polytrichia, and Setosa can be discounted because the fossil lacks subulate scales or scales with acicular marginal hairs. Thus, the fossil might belong to either sects. Amygdalifolia, Wrightiana, Elaphoglossum, or Squamipedia or to an extinct lineage.• CONCLUSIONS: The discovery of a Miocene Elaphoglossum fossil provides remarkable support to current molecular clock-based estimates of the diversification of these ferns.

Phylogeny and historical biogeography of the lastreopsid ferns (Dryopteridaceae).

• Premise of the study: As currently circumscribed, Lastreopsis has about 45 species and occurs in Australia, southern Asia, Africa, Madagascar, and the neotropics. Previous molecular phylogenetic studies suggested that Lastreopsis is paraphyletic. Our study focuses on resolving relationships among the lastreopsid ferns (Lastreopsis, Megalastrum, and Rumohra), the evolution of morphological characters, and an understanding of the temporal and spatial patterns that have led to the current diversity and geographical distribution of its extant species.• Methods: Phylogenetic relationships were recovered under Bayesian, maximum likelihood, and maximum parsimony methods, using a data set of four plastid markers. Divergence time estimates were made using BEAST, and the biogeographic hypotheses were tested under the DEC model and the RASP/S-DIVA methods.• Key results: Lastreopsis was recovered as paraphyletic, and at least one of its clades should be recognized as a distinct genus, Parapolystichum. Coveniella poecilophlebia and Oenotrichia tripinnata were nested within Lastreopsis s.s., Megalastrum and Rumohra as sister to the Lastreopsis s.s., and the Lastreopsis amplissima clades. The initial diversification of the lastreopsids took place at about 56.55 Ma, from a neotropical ancestor.• Conclusions: Taxonomic recognition of Parapolystichum is warranted to preserve the monophyly of Lastreopsis. Diversification among the main clades of the lastreopsid ferns was influenced by climatic and geological changes in the southern hemisphere. The biogeographic history of the group is intimately related to the trans-Antarctic corridor between Australia and South America, with evidence for multiple lineage interchanges between Australia and South America during the Oligocene and the Eocene epochs.

The evolution, morphology, and development of fern leaves.

Leaves are lateral determinate structures formed in a predictable sequence (phyllotaxy) on the flanks of an indeterminate shoot apical meristem. The origin and evolution of leaves in vascular plants has been widely debated. Being the main conspicuous organ of nearly all vascular plants and often easy to recognize as such, it seems surprising that leaves have had multiple origins. For decades, morphologists, anatomists, paleobotanists, and systematists have contributed data to this debate. More recently, molecular genetic studies have provided insight into leaf evolution and development mainly within angiosperms and, to a lesser extent, lycophytes. There has been recent interest in extending leaf evolutionary developmental studies to other species and lineages, particularly in lycophytes and ferns. Therefore, a review of fern leaf morphology, evolution and development is timely. Here we discuss the theories of leaf evolution in ferns, morphology, and diversity of fern leaves, and experimental results of fern leaf development. We summarize what is known about the molecular genetics of fern leaf development and what future studies might tell us about the evolution of fern leaf development.

Nuclear genome size in Selaginella.

Estimates of nuclear genome size for 9 Selaginella species were obtained using flow cytometry, and measurements for 7 of these species are reported for the first time. Estimates range from 0.086 to 0.112 pg per holoploid genome (84-110 Mb). The data presented here agree with the previously published flow cytometric results for S. moellendorffii. Within the 9 species sampled here, chromosome number varies from 2n = 16 to 2n = 27. Nuclear genome size appears to be strongly correlated with chromosome number (Spearman's rank correlation; p = 0.00003725). Cultivated S. moellendorffii lacks sexual reproduction--manifest by the production of abortive megasporangia. Flow cytometric data generated from a herbarium specimen of a fertile wild-collected S. moellendorffii are virtually indistinguishable from the data generated from fresh material (0.088 vs. 0.089 pg/1C). Therefore, the limited fertility observed in cultivated plants is probably not the result of abnormal chromosome number (e.g., induced by interspecific hybridization).

Molecular phylogeny of the fern genus Elaphoglossum (Elaphoglossaceae) based on chloroplast non-coding DNA sequences: contributions of species from the Indian Ocean area.

We performed a phylogenetic analysis of the fern genus Elaphoglossum using two non-coding chloroplast spacers: trnL-trnF and rps4-trnS. The sampling includes 123 species, of which 80 have not been previously sequenced, and for the first time includes species from Africa and the Indian Ocean area. The results of this expanded study largely agree with an earlier molecular study based on a smaller group of neotropical species and with the morphology-based classification of Mickel and Atehortua. We found, however, that some infrageneric groups such as section Elaphoglossum are not monophyletic. Besides section Elaphoglossum pro parte, we recognize six sections: two new monospecific, unnamed sections, and the previously established sections Lepidoglossa, Squamipedia, Amygdalifolia, and "Subulate-scaled clade." We divide the subulate-scaled clade into subsection Setosa (hydathodes present) and Polytrichia (hydathodes absent), and section Elaphoglossum is divided into subsections Platyglossa and Pachyglossa, two groups that do not appear to be supported by any single morphological character. In general, however, the main clades are supported by morphology. Finally, we discuss the species of the Indian Ocean region and their affinities with the neotropical ones. Out of the 11 species pairs postulated by Moran and Smith on the basis of morphology, two are well supported (E. eximium-E. aubertii; E. piloselloides-E. spatulatum) and three are not supported (E. ciliatum-E. humbertii; E. muscosum-E. poolii; E. paleaceum-E. deckenii), and two remain unresolved (E. erinaceum-E. hybridum; E. glabellum-E. acrostichoides) because our molecular markers were not variable enough. Four species pairs could not be tested because specimens were lacking. Unsupported species pairs are best interpreted as morphological convergences. Two additional species pairs are proposed: E. cuspidatum-E. succisaefolium; E. doanense-E. hornei. Placement of the species from the Indian Ocean suggests that at least 13 long-distance dispersal events occurred between the Neotropics and the Indian Ocean-Africa.