Historical biogeography of the fern genus Polystichum (Dryopteridaceae) in Austral South America.

A group of seven endemic Polystichum species inhabit Patagonia, the southern region of South America. To date, evolutionary relationships of these Austral South American Polystichum remain unknown. The biota of the Southern Andes appears to be more closely related to the temperate Australasian species than to northern South American ones. Differences in morphological characters suggested that Austral South American Polystichum follows that biogeographical pattern, not being closely related to their congeners in the Northern and Central Andes. We sought to reveal the evolutionary relationships, estimate the divergence times and reconstruct both ancestral areas and ancestral ploidy levels of Austral South America Polystichum. Phylogenetic relationships were estimated using maximum likelihood and Bayesian approaches. The seven Austral South American species plus 31 Polystichum species spanning all other major biogeographic regions were sampled for three DNA markers. Divergence times were estimated in BEAST and Bayesian binary Markov chain Monte Carlo reconstruction was applied in order to infer ancestral areas. The evolution of ploidy was reconstructed on the maximum clade credibility tree, using stochastic character mapping. Austral South American Polystichum was recovered as monophyletic. The earliest divergence reconstructed within the Austral South American Clade was that of Polystichum andinum; subsequently two other lineages diverged comprising the remaining Austral South American species. The Austral South American lineage is not closely allied to North and Central Andes congeners. Long-distance dispersal of an ancestral tetraploid from Australasia during the late Miocene is the most likely explanation for the origin of Patagonian Polystichum. Then, Pliocene and Pleistocene orogenic and climatic changes may have shaped its diversification in Patagonia.

Squalene Cyclases and Cycloartenol Synthases from Polystichum polyblepharum and Six Allied Ferns.

Ferns are the most primitive of all vascular plants. One of the characteristics distinguishing them from flowering plants is its triterpene metabolism. Most cyclic triterpenes in ferns are hydrocarbons derived from the direct cyclization of squalene by squalene cyclases (SCs). Both ferns and more complex plants share sterols and biosynthetic enzymes, such as cycloartenol synthases (CASs). Polystichum belongs to Dryopteridaceae, and is one of the most species-rich of all fern genera. Several Polystichum ferns in Japan are classified as one of three possible chemotypes, based on their triterpene profiles. In this study, we describe the molecular cloning and functional characterization of cDNAs encoding a SC (PPH) and a CAS (PPX) from the type species Polystichum polyblepharum. Heterologous expression in Pichia pastoris revealed that PPH and PPX are hydroxyhopane synthase and CAS, respectively. By using the PPH and PPX sequences, we successfully isolated SC- and CAS-encoding cDNAs from six Polystichum ferns. Phylogenetic analysis, based on SCs and oxidosqualene cyclase sequences, suggested that the Polystichum subclade in the fern SC and CAS clades reflects the chemotype-but not the molecular phylogeny constructed using plastid molecular markers. These results show a possible relation between triterpenes and their biosynthetic enzymes in Polystichum.

Insights into evolution in Andean Polystichum (Dryopteridaceae) from expanded understanding of the cytosolic phosphoglucose isomerase gene.

Cytosolic phosphoglucose isomerase (pgiC) is an enzyme essential to glycolysis found universally in eukaryotes, but broad understanding of variation in the gene coding for pgiC is lacking for ferns. We used a substantially expanded representation of the gene for Andean species of the fern genus Polystichum to characterize pgiC in ferns relative to angiosperms, insects, and an amoebozoan; assess the impact of selection versus neutral evolutionary processes on pgiC; and explore evolutionary relationships of selected Andean species. The dataset of complete sequences comprised nine accessions representing seven species and one hybrid from the Andes and Serra do Mar. The aligned sequences of the full data set comprised 3376 base pairs (70% of the entire gene) including 17 exons and 15 introns from two central areas of the gene. The exons are highly conserved relative to angiosperms and retain substantial homology to insect pgiC, but intron length and structure are unique to the ferns. Average intron size is similar to angiosperms; intron number and location in insects are unlike those of the plants we considered. The introns included an array of indels and, in intron 7, an extensive microsatellite array with potential utility in analyzing population-level histories. Bayesian and maximum-parsimony analysis of 129 variable nucleotides in the Andean polystichums revealed that 59 (1.7% of the 3376 total) were phylogenetically informative; most of these united sister accessions. The phylogenetic trees for the Andean polystichums were incongruent with previously published cpDNA trees for the same taxa, likely the result of rapid evolutionary change in the introns and contrasting stability in the exons. The exons code a total of seven amino-acid substitutions. Comparison of non-synonymous to synonymous substitutions did not suggest that the pgiC gene is under selection in the Andes. Variation in pgiC including two additional accessions represented by incomplete sequences provided new insights into reticulate relationships among Andean taxa.

Using a multilocus phylogeny to test morphology-based classifications of Polystichum (Dryopteridaceae), one of the largest fern genera.

BACKGROUND: Polystichum (Dryopteridaceae) is probably the third largest fern genus in the world and contains ca. 500 species. Species of Polystichum occur on all continents except Antarctica, but its highest diversity is found in East Asia, especially Southwest China and adjacent regions. Previous studies typically had sparse taxon sampling and used limited DNA sequence data. Consequently, the majority of morphological hypotheses/classifications have never been tested using molecular data. RESULTS: In this study, DNA sequences of five plastid loci of 177 accessions representing ca. 140 species of Polystichum and 13 species of the closely related genera were used to infer a phylogeny using maximum likelihood, Bayesian inference, and maximum parsimony. Our analyses show that (1) Polystichum is monophyletic, this being supported by not only molecular data but also morphological features and distribution information; (2) Polystichum is resolved into two strongly supported monophyletic clades, corresponding to the two subgenera, P. subg. Polystichum and P. subg. Haplopolystichum; (3) Accessions of P. subg. Polystichum are resolved into three major clades: clade K (P. sect. Xiphophyllum), clade L (P. sect. Polystichum), and the HYMASO superclade dominated by accessions of P. sect. Hypopeltis, P. sect. Macropolystichum, and P. sect. Sorolepidium, while those of P. subg. Haplopolystichum are resolved into eight major clades; and (4) The monophyly of the Afra clade (weakly supported), the Australasian clade (weakly supported), and the North American clade (strongly supported) is confirmed. CONCLUSIONS: Of the 23 sections of Polystichum recognized in a recent classification of the genus, four (P. sect. Hypopeltis, P. sect. Neopolystichum, P. sect. Sorolepidium, P. sect. Sphaenopolystichum) are resolved as non-monophyletic, 16 are recovered as monophyletic, and three are monospecific. Of the 16 monophyletic sections, two (P. sect. Adenolepia, P. sect. Cyrtogonellum) are weakly supported and 14 are strongly supported as monophyletic. The relationships of 11 sections (five in P. subg. Haplopolystichum; six in P. subg. Polystichum) are well resolved.

Phylogeny and biogeography of exindusiate Andean Polystichum (Dryopteridaceae).

PREMISE OF THE STUDY: Uplift of the tropical Andes had a significant impact on the diversification of South American flora and fauna. Recent biogeographic inquiries have established patterns of Andean divergence, but investigations on ferns are scant. The fern genus Polystichum Roth (Dryopteridaceae) combines widespread geographic and elevational distribution with a large number of species to form an ideal system for investigation of the origin and diversification patterns of a fern lineage in the tropical Andes. METHODS: The relationships among 42 Polystichum species, including taxa from all major biogeographic regions, were analyzed with 2591 aligned nucleotides from four plastid markers using maximum parsimony and Bayesian inference. The resulting phylogeny was then used to estimate divergence times and reconstruct both ancestral areas and ancestral elevations. KEY RESULTS: Tropical Andean South American polystichums that lack an indusium (sori exindusiate) were confirmed to form a monophyletic group. This exindusiate Andean Polystichum clade diverged from a middle-elevation forest lineage now rich in species endemic to Mexico during the middle Miocene (13.12 million years ago). The majority of diversification that followed took place in the montane regions of the central Andes with radiations to the northern Andes, southeastern Brazil, and alpine regions. CONCLUSIONS: The monophyletic exindusiate Andean Polystichum lineage diverged from a Mexican lineage in the middle Miocene and diversified in the central Andes before dispersing northward. This south-to-north dispersal pattern, documented for many other Andean lineages, corresponds with episodes of uplift in the tropical Andes.

Variation in ploidy level and phenology can result in large and unexpected differences in demography and climatic sensitivity between closely related ferns.

PREMISE OF THE STUDY: Current environmental changes may affect the dynamics and viability of plant populations. This environmental sensitivity may differ between species of different ploidy level because polyploidization can influence life history traits. We compared the demography and climatic sensitivity of two closely related ferns: the tetraploid Polystichum aculeatum and one of its diploid parents, Polystichum setiferum. METHODS: Matrix models were used to assess the effects of life history variation on population dynamics under varying winter conditions. We analyzed the contributions of all key aspects of the fern life cycle to population growth. Our study is the first to also include the gametophyte generation. KEY RESULTS: Projected population growth rate (λ) was much higher for the tetraploid P. aculeatum (1.516) than for P. setiferum (1.071) under normal winter conditions. During a year with harsh winter conditions, population growth of P. aculeatum was strongly reduced. This finding contradicts our expectation that the winter-hardy fronds of this species would allow high survival of harsh winters. Differences in λ between species and between years with different winter conditions were mostly caused by variation in gametophyte-related recruitment rates, a finding that shows the importance of including gametophytes in fern demographic studies. CONCLUSIONS: Our results indicate that populations of closely related ferns can show large differences in population performance, mainly related to recruitment rates and frond phenology, and that these differences may depend greatly on climatic conditions. Our findings provide a first indication that (allo)polyploidization in ferns can have a significant effect on population dynamics.

Polyphyly of the fern family Tectariaceae sensu Ching: insights from cpDNA sequence data.

Tectariaceae are a pantropical fern family of about 20 genera, among which 8 are distributed in China. The morphological distinctiveness of the family is widely recognized, yet relatively little systematic research has been conducted on members of Tectariaceae. Phylogenetic analyses of chloroplast DNA sequence data (rbcL and atpB) from 15 species representing all 8 genera in China were carried out under parsimony criteria and Bayesian inference. The phylogenetic reconstructions indicated that the fern family Tectariaceae as traditionally circumscribed are polyphyletic. Ctenitis, Dryopsis, Lastreopsis clustered with and should be included within the newly-defined Dryopteridaceae, and Pleocnemia is also tentatively assigned to it. A narrowly monophyletic Tectariaceae is identified, which includes Ctenitopsis, Hemigramma, Pteridrys, Quercifilix, and Tectaria. In the single rbcL analysis, Arthropteris clustered with the above-mentioned monophyletic Tectariaceae. Although further investigations are still needed to identify infrafamilial relationships within the monophyletic Tectariaceae and to redefine several problematic genera, we propose a working concept here that better reflects the inferred evolutionary history of this group.