Quantitative expression of fern leaf development and fertility inPolystichum tripteron (Aspidiaceae)

To elucidate the quantitative and qualitative expression of fern development, various features of leaf-size and morphology were examined on leaves ofPolystichum tripteron (Aspidiaceae) in Japan. Among ten parameters, the number of branches of the leaf midrib (NV) proved to be the most successful for discrimination between fertile and sterile leaf populations. In the phase-change from sterile to fertile leaves, the NV parameter also gave the strongest correlation, increasing with fertility (r = 0.983), and is therefore a useful tool for research into the life history characteristics of ferns.Contribution no. 2751 from the Institute of Low Temperature Science, Hokkaido University.     

A quantitative comparison of foliage development among allopatric ferns,Dryopteris crassirhizoma,D. coreano-montana andD. filix-mas

A quantitative comparison was conducted on the foliage development during sporophyte development of three allopatric ferns in cool temperate and subalpine regions of Hokkaido and Tirol, European Alps. The foliage development ofDryopteris crassirhizoma, D. coreano-montana andD. filix-mas was quantitatively described by the leaf development (NV, number of veins); NV correlates the leaf-shape complexity from a circle (DI, L/2(3.14×S)^1/2). Nearly similar patterns were detected on frequency distribution of fertile leaves, fertility increase and number of leaves in threeDryopteris ferns which exhibit funnel-shaped foliage arrangements in mature sporophyte. No difference was found in number of leaves, maximum NV, fertility rate and leaf-shape parameters among three ferns. A positive difference was found only on changes in order of pinnae with maximum number of costa branches (NVP) and the DI of outline of pinnae betweenD. crassirhizoma andD. filix-mas. These allopatricDryopteris ferns seem to have a similar foliage structure, in spite of some sympatricDryopteris ferns capable of producing putative hybrids (D. austriaca andD. amurensis; D. tokyoensis andD. monticola) having different foliage structures in Hokkaido. Contribution No. 3346 from the Institute of Low Temperature Science, Hokkaido University.     

Life history characteristics ofPolystichum tripteron with special reference to its leaf venation

An intra-species comparison of the life history of various populations ofPolystichum tripteron was undertaken in Hokkaido and Honshu in Japan with respect to some leaf-size characters and fertility during sporophyte development. The developmental process of sporophytes was quantitatively estimated by counting the number of venation (NV: number of midrib branches of the leaf). The mean NV of the fertile leaves decreased slightly with increasing altitude on Mt. Teine near Sapporo and with increasing latitudes from Honshu to Hokkaido. The blade length was longer at any NV in Hokkaido populations than in those of southern Honshu. Longer blade (BL) and pinna length (PL) and lesser number of pinnules of the lowest pinna (NPLP) versus NV were observed in leaf populations from Hokkaido than in those from southern Honshu. The maximum values of lowest pinna length (LPL), NV, number of costa branches (NVLP) and number of pinnule (NPLP) were larger in southern Honshu than in Hokkaido. In the course of leaf-shape formation represented by the ratio of PL to BL versus NV, the Kyoto population seemed to be more differentiated into long-lanceolate fronds than did the Hokkaido population. Initiation of fertility in the sporophyte development and the maximum NV seemed to decrease with increasing altitudes and latitudes in Japan. Contribution No. 2885 from the Institute of Low Temperature Science, Hokkaido University.     

Comparative life history of aspidiaceous ferns in northern Japan with reference to fertility during sporophyte development in relation to habitats

The life-cycle characteristics of ferns, in particular reproductive characteristics in relation to alternation of generations, have been studied in populations of 19 species of aspidiaceous ferns native to Hokkaido. The developmental age of the sporophyte was quantitatively expressed by the number of midrib branches of the leaf (NV; number of venation). The maximum developmental age (MNV) of the aspidiaceous ferns in Hokkaido ranged from 49 to 113. The initiation of fertility (IF) in the relative developmental age (RDA), percentages of any NV to the maximum NV of the leaf population) ranged from 15 to 65%. Ferns occurring on rocky cliffs initiated fertility at 15–21%, while ferns occurring on the forest floor initiated fertility at 41–65% in the RDA of a leaf population. Further observations were undertaken on 26 ferns of other families and revealed that there were correlations between habitat and maximum NV, and habitat and IF in the RDA. Ferns occurring on cliffs, on mossy rocks and on tree trunks, had lower maximum NV of the leaf and earlier initiation of fertility in the RDA, than those of ferns occurring on the forest floor, edge of forests and on grassland in Hokkaido. Ferns occurring in the exposed habitat seemed to behave as a r-strategist, while ferns occurring in the forest floor seemed to behave as a K-strategist.     

Quantitative comparison of foliage development amongDryopteris monticola,D. tokyoensis and a putative hybrid,D. kominatoensis in Northern Japan

Developmental leaf architecture was quantitatively described in terms of measurements of various parameters on leaf blade from different size of sporophytes inDryopteris monticola, D. tokyoensis and a putative hybrid,D. kominatoensis in the natural site of Hokkaido, to compare the ontogenetic differentiation in foliage structure among allied ferns. The morphological stage of leaf and sporophyte was tentatively quantified by the number of midrib branches of the leaf (NV, number of veins), which exhibited a significant correlation to the leaf-shape complexity from a circle (DI=marginal length/2×(3.14×square)^1/2) of leaf blade. D. kominatoensis showed intermediate values between others in following characters; DI increase, maximum NV (also blade length), maximum number of costa branches of pinnae (NVMP), number of costa branches of the lowest pinna (NVLP), difference between NVMP and NVLP (NVMP-NVLP), during heteroblastic leaf development. A larger number of leaves per sporophyte was found inD. kominatoensis than in others. The fertility rate (%) and initiation of fertility (IF) in the relative developmental stage (RDS) ofD. kominatoensis shifted to that ofD. tokyoensis, while the order of pinnae with NVMP shifted to that ofD. monticola. Even in the intermediate characters inD. kominatoensis, slight shifts in characters to those of putative parents were found during heteroblastic leaf development. Contribution No. 3145 from the Institute of Low Temperature Science, Hokkaido University.     

Life history characteristic ofCyrtomium falcatum around the natural northern boundary in Hokkaido,with reference to the alternation of generations

To understand the life history characteristic for expanding the distributional area to colder climates, developmental age structure of population ofCyrtomium falcatum was observed along southwestern coasts of Hokkaido at the natural northern boundary of its distribution, with reference to the alternation of generations. The length and number of pinna of fertile leaves ofCyrtomium falcatum decrease towards the northern part of Japan. In southwestern Hokkaido, typically dwarf fertile leaves and gametophytes were observed growing together on cliffs nearby the sea. To estimate the developmental ages of small and dwarf leaves, the number of venation (NV: branching number of vein from midrib) of leaves was counted on each sporophyte. The sporophyte with leaves at the simple pinna stage ranging from 0–25NV, is predominant in the population of southwestern coasts of Hokkaido. The fertility of the sporophyte seems to be achieved more than five years after the germination. The gametophytes were also observed at the location to be almost equal in number to sporophytes. The number of gametophytes and sporophytes decreases with advancement of developmental stages. In the same location at Okushiri Isl. with slight gradiency of humidity, the gametophyte is predominant on the drier cliff, while the sporophyte is predominant on the humid hole. The population ofCyrtomium falcatum at the natural northern boundary in Hokkaido, seems to have the life history characteristic with alternation of generations. Contribution No. 2557 from the Inst. of Low Temp. Sci.     

Conservation of rDNA inPolystichum (Dryopteridaceae)

Restriction site variation in the nuclear 18S–25S ribosomal RNA genes (rDNA) was analyzed hierarchically in a species complex in the fern genusPolystichum. Two distinct rDNA repeat types were present in all individuals ofPolystichum examined. No variation was detected among individuals within a population ofP. munitum, among populations ofP. munitum orP. imbricans, or among the six diploid species ofPolystichum from North America, including the circumborealP. lonchitis. The identity of rDNA repeats across all six North American species ofPolystichum may reflect an overall similarity of the nuclear genomes of these species, an observation supported by isozyme data as well. However, this nuclear similarity contrasts sharply with the highly divergent chloroplast genomes of these six species. The conservative nature of the rDNA inPolystichum also is in contrast to the much more variable rDNAs of most angiosperms investigated. Perhaps the tempo and mode of evolution of rDNA in ferns differ from those of angiosperms; however, the data base for fern rDNA is very small. Furthermore, the number of repeat types per individual is consistent with a diploid, rather than polyploid, condition despite the high chromosome number (n = 41) of these plants, although homogenization of multiple, divergent rRNA genes cannot be disproven.     

Estimation of chronological age for sporophyte maturation in three semi-evergreen ferns in Hokkaido

To elucidate the life cycle time schedule of semi-evergreen ferns at natural sites in Sapporo, the chronological age for sporophyte maturation was estimated from the increase in the number of veins (NV, number of midrib branches) from overwintered to newly expanded leaves. The measurements were conducted on populations ofPolystichum braunii, Polystichum tripteron andDryopteris crassirhizoma at altitudes of 50–650 m on Mt. Teine to compare the interspecific maturation age of sporophytes. The mean age for sporophyte maturation was estimated to be 7.6 yr forP. braunii, 16.5 yr forP. tripteron and 12.6 yr forD. crassirhizoma. The minimum ages (1% fertility rate) for sporophyte maturation ofP. braunii, P. tripteron andD. crassirhizoma were 5.8, 11.0 and 7.9 yr, respectively. The maturation age was shortest forP. braunii and longest forP. tripteron. The maturation age of a species thus seems to differ according to altitude and habitat, and that of the present three species seems to be influenced by the geographical location of the species distribution. Contribution No. 3315 from the Institute of Low Temperature Science, Hokkaido University.     

Characteristics of “Number of Veins” to Estimate Leaf Maturity in Pteris mutilata (Pteridaceae)

Pteris mutilata , as has been established in ferns of cool temperate regions. NV expressed developmental stages better than any other leaf size parameters, such as blade length, blade width, stipe length, and total length (blade+stipe length). The leaf shape became more oblong and/or slender after the plant matured, which could be measured by two shape parameters, (blade width)/(total length) and (blade width)/(blade length). Principal component analysis using all the morphological parameters showed that NV is categorized into size parameters, although NV has been considered to differ somehow from the other size parameters. Thus NV represents one of the size parameters that is the most appropriate to estimate leaf maturation. Received 6 July 2000/ Accepted in revised form 22 August 2000     

A quantitative estimation of sequential foliage development and fertility inDryopteris crassirhizoma

The foliage development ofDryopteris crassirhizoma Nakai was quantitatively estimated by measurements of shape and size of leaves from different developmental stages of sporophytes, to lead to an understanding of the life history characteristics of the species. The number of midrib branches (NV, number of veins) of the leaf corresponds to the leaf-shape complexity (DI, dissection index; shape complexity from a circle) and length of leafblade (BL). Some quantitative characters, such as leaf uniformity (decrease in NV variation), changes in shape and increase in number of leaves, vary progressively during foliage formation. The sequence of foliage development can be quantified using the parameter NV: for example, 15-NV for leaf uniformity, 30-NV for leaf-shape change from triangular to oblanceolate, 60-NV for increase in leaf number and leaf fertility in the course of sporophyte ontogeny. Contribution No. 3297 from the Institute of Low Temperature Science, Hokkaido University.     

Phylogeny and biogeography of Chinese and Australasian Polystichum ferns as inferred from chloroplast trnL-F and rps4-trnS sequence data

Polystichum, one of the largest genera of ferns, occurs worldwide with the greatest diversity in southwest China and adjacent regions. Although there have been studies of Chinese Polystichum on its traditional classification, geographic distributions, and even a few on its molecular systematics, its relationships to other species outside China remain little known. Here, we investigated the phylogeny and biogeography of the Polystichum species from China and Australasia. The evolutionary relationships among 42 Polystichum species found in China (29 taxa) and Australasia (13 taxa) were inferred from phylogenetic analyses of two chloroplast DNA sequence data sets: rps4-trnS and trnL-F intergenic spacers. The divergence time between Chinese and Australasian Polystichum was estimated. The results indicated that the Australasian species comprise a monophyletic group that is nested within the Chinese diversity, and that the New Zealand species are likewise a monophyletic group nested within the Australasian species. The divergence time estimates suggested that Chinese Polystichum migrated into Australasia from around 40 Ma ago, and from there to New Zealand from about 14 Ma. The diversification of the New Zealand Polystichum species began about 10 Ma. These results indicated that Polystichum probably originated in eastern Asia and migrated into Australasia: first into Australia and then into New Zealand.     

Frond development and phenology of Thelypteris limbosperma, Athyrium distentifolium, and Matteuccia struthiopteris in Western Norway

Odland, A. 1995. Frond development and phenology of Thelypteris limbosperma, Athyrium distentifolium, and Matteuccia struthiopteris in Western Norway. — Nord. J. Bot. 15: 225–236. Copenhagen. ISSN 0107–055X. The pattern of growth and phenology of Thelypteris limbosperma, Athyrium distentifolium, and Matteuccia struthiopteris fronds has been investigated, with particular emphasis on height increments of the sporophytes and the development of fertile fronds. In order to study both interspecific and intraspecific differences, fern stands along altitudinal gradients have been monitored. To investigate the initiation of fertility, fronds representing different developmental stages have been sampled. Climatic data have been collected with a data-logger during the study periods. The ferns show significant differences in growth pattern and phenology. Matteuccia struthiopteris has a growth curve that is best described as monomolecular, characterised by high growth rates during the early developmental stage, while the development of Thelypteris limbosperma and Athyrium distentifolium follow a logistic growth curve. The species need different periods of time to develop their fronds. The main frond elongation does not start before soil temperature has reached 7 C. After that, the growth is mainly controlled by air temperature. Within the interval when mean maximum temperatures are 8–20 C, all three ferns showed increased growth rates with increasing air temperatures, but the increase was greatest in M. struthiopteris. At higher temperatures, the growth rate of M. struthiopteris and T. limbosperma decreased. Athyrium distentifolium is characterised by producing sori at an early developmental stage. Fertile Matteuccia strurhiopteris fronds are developed at a much later developmental stage. Thelypteris limbosperma needs a longer period of time to produce mature fronds than the other species. The investigation indicates that plant growth and development are determined by both internal and external factors. It is concluded that growth rate, phenology, and the ability of ferns to produce mature fronds reveal close similarities with their response to environmental variables, and hence with their broad-scale geographical distributional patterns.     

Phenology and wintering capacity of sporophytes and gametophytes of ferns native to northern Japan

Summary To investigate life history adaptations to cold climates, the leaf development, sporulation period, growing stage of gametophytes, and the frost and drought resistance of sporophytes and gametophytes of 67 fern species native to Kokkaido were studied. Most ferns common in Hokkaido are summer-green with leaves developing during late May to June and decaying during October. Most of the ferns in Hokkaido sporulate during August to early September. Spores dispersed from June to September germinate before winter begins, forming vegetative prothallia. Gametophytes mature only in the following summer. Thus in Hokkaido the gametophytes as well as perennial sporophytes are exposed to severe winter conditions. In order to correlate the life cycles of temperate ferns with winter cold stress, frost resistance of gametophytes, rhizomes, and leaves of sporophytes were determined. Maximal frost resistance of rhizomes reflects the stress conditions of their habitats: rhizomes of forest understory ferns are damaged at-5°to -17.5°C, epiphytic ferns and ferns of habitats exposed to severe frost sustained temperatures of -20° to-40°C. The leaves of winter-green and evergreen ferns resist frost ranging from -25° to -40°C. The leaves of summer-green ferns are killed by late frost below -5°C. With some exceptions, gametophytes of ferns growing on the forest floor resist frost to -40°C and are much hardier than sporophytes. These results suggest the possible restrictive effects of cold climate on the life span of leaves as well as on the sporulation period. If winter cold is one of the decisive factors for seasonality expression and habitat distribution of ferns, the sensitive generation must be the sporophyte rather than the gametophyte. The hardier gametophyte is therefore able to colonize habitats in which the sporophyte is excluded by frost if mechanisms of vegetative propagation are evolved.Contribution No. 2451 from The Institute of Low Temperature Science     

Foliar water uptake,a widespread phenomenon in temperate woodland ferns?

Most woodland ferns thrive under conditions of high air humidity, frequent precipitation and exposure to extended periods of leaf wetness, but it is not known how widespread foliar water uptake is in this plant group. In a tracer experiment with deuterated water (²H₂O) applied to the leaf surface of five temperate woodland ferns (Athyrium filix-femina, Dryopteris filix-mas, Polystichum aculeatum, Polystichum braunii and Asplenium scolopendrium), we tested (1) whether these species exhibit foliar water uptake and (2) whether the capability to absorb water through the leaf epidermis increases with the frequency of epidermal trichomes. All species had significantly higher abundances of ²H in tissue water, when extracted distant to the place of application, compared to the background level (0.052–0.504 vs. 0.015 at.%), evidencing uptake through the epidermis and leaf-internal translocation. A positive relation between trichome density and ²H incorporation was found only for the second-order pinnae but not for the more central frond sections. The results suggest that foliar water uptake may be widespread among temperate woodland ferns across different families and that leaf trichome structure probably influences this process.     

苦竹叶片碳氮磷化学计量特征的海拔梯度效应

为揭示海拔梯度对苦竹林立竹叶片碳(C)、氮(N)、磷(P)化学计量特征的影响,该文以3个海拔梯度[低海拔,(200±10) m;中海拔,(400±10) m;高海拔,(800±10) m]苦竹林为研究对象,测定1至3年生立竹叶片C、N、P含量,分析其化学计量特征和异速增长关系。结果表明:(1)立竹年龄对苦竹叶片C、N、P含量及其化学计量特征影响明显,随立竹年龄的增大,苦竹叶片C、N、P含量和N∶P总体上均呈降低趋势,而C∶N、C∶P总体上呈升高趋势。(2)海拔对苦竹叶片C、N、P含量及其化学计量特征有重要影响,随海拔梯度升高,不同年龄立竹叶片C含量呈先升高后下降变化趋势,N、P含量总体上呈降低趋势,而C∶N、C∶P和N∶P均呈升高趋势。(3)不同海拔梯度苦竹林立竹叶片C、N、P含量和C∶N、C∶P差异显著,中、高海拔苦竹林立竹叶片N∶P无显著差异,均显著高于低海拔苦竹林;不同海拔梯度苦竹林立竹叶片C、N、P间呈显著的正异速增长关系,随海拔梯度的升高,C-N、C-P异速增长指数显著升高,而N-P异速增长指数显著下降。综上结果表明,高海拔苦竹林虽然N、P利用效率提高,但立竹叶片C含量较低,P限制性作用增强;中海拔苦竹林不但具有较高的N、P利用效率,而且立竹叶片C含量高,说明中海拔是苦竹林丰产培育的适宜海拔。     

Phylogeny of Chinese <Emphasis Type="Italic">Polystichum</Emphasis> (Dryopteridaceae) based on chloroplast DNA sequence data (<Emphasis Type="Italic">trnL-F</Emphasis> and <Emphasis Type="Italic">rps4-trnS</Emphasis>)

Polystichum is one of the largest and most taxonomically complex fern genera in China. The evolutionary relationships of Chinese Polystichum and related genera, and the relationship between our Polystichum phylogeny and ecogeographic distribution, were tested by the use of DNA sequence data. Fifty-one species of Polystichum and 21 species in allied genera were sequenced for the plastid intergenic spacers rps4-trnS and trnL-F. Maximum parsimony and Bayesian phylogenetic analyses of both individual and combined data sets showed that Chinese Polystichum as commonly recognized was paraphyletic: one clade (the CCPC clade) included Cyrtomidictyum lepidocaulon, two Cyrtogonellum species, three Cyrtomium species, and a small number of Polystichum species usually occurring on limestone. A second clade, Polystichum sensu stricto, included the remainder of the Polystichum species; these often occur on non-limestone substrates. The remaining Cyrtomium species formed the third clade. Three subclades resolved within Polystichum sensu stricto (s.s.) clade do not correspond with recent sectional classifications, and we outline the issues relevant to a new classification for the genus. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Fern-associated arbuscular mycorrhizal fungi are represented by multiple Glomus spp.: do environmental factors influence partner identity?

Symbioses involving arbuscular mycorrhizal fungi (AMF) are among the most important ecological associations for many plant species. The diversity of AMF associated with ferns, however, remains poorly studied. Using recently designed Glomus-specific primers, we surveyed the AMF community associated with ferns from deciduous, broad-leaved second-growth forest habitats at the eastern edge of the piedmont region of central Virginia, USA. Results indicate that this molecular approach may be a useful tool for detecting AMF in ferns compared to traditional techniques based on morphology. Over 30 potential fungal ribotypes were identified from eight fern species using denaturing gradient gel electrophoresis. Fungal ribotypes were found to differ widely in terms of (1) the number of fern partners with which they interact and (2) their relative frequency within each fern. Sequence analysis of fungal isolates from three species of fern indicated that the primers were generally highly specific for Glomus species but some non-target DNA was also amplified. Cloned polymerase chain reaction (PCR) products from Polystichum acrostichoides and Osmunda regalis revealed several phylogenetically distinct Glomus species. A single Glomus species was identified in the cloned PCR products from Botrychium virginianum. These findings challenge the hypothesis that the extent or degree of fern–fungal symbiosis is somehow tied to root complexity. Environmental factors appear to influence the suite of AMF that form partnerships with ferns. Some species of fern from similar habitats associated with dissimilar fungal partners (e.g., P. acrostichoides and Athyrium filix-femina var. asplenioides), whereas others harbored uniform fungal communities (e.g., Asplenium platyneuron). The significance of these data in terms of ecological and evolutionary dynamics of the AMF–fern symbiosis is discussed. Brittany West, Jessica Brandt, and Kay Holstien contributed equally to this work.     

Roles for structural and temporal shelter-changing by fern-feeding lepidopteran larvae

Summary Larvae of the pyralid moth, Herpetogramma aeglealis, construct feeding shelters upon the Christmas fern (Polystichum acrostichoides). Field and laboratory study involving 532 shelters showed that as the larvae mature, they sequentially inhabit approximately 5 shelters of 3 distinct types, constructed at night on different fronds of the same plant. The bundle shelter, simple and ephemeral, is first to be inhabited and constructed. The fiddlehead shelter which houses slightly older larvae strongly resembles contemporaneously emerging frond fiddleheads. The final shelter form, the globe, is a silk-bound ball of leaflets at the frond tip. An individual larva usually constructs 3 globe shelters on different fronds of the same plant before completing its development. As shelter sites, sterile Polystichum fronds are chosen preferentially over fertile fronds. The bundle and fiddlehead shelter forms, less abundant, appear cryptic to humans and perhaps to other vertebrates. The final globe shelter form is larger and quite conspicuous. However, the persistence of empty globe shelters left on the plant as the larva moves to a new one may serve to make searching for larvae less profitable for potential predators and parasites. We suggest that the energetic costs of constructing and occupying multiple shelters may be offset by circumvention of reduced frond palatability and reduced exposure to predators and parasites.     

陕西太白山自然保护区蕨类植物区系研究

对太白山自然保护区蕨类植物区系进行了初步的研究。结果表明:太白山共有蕨类植物21科、39属、110种,并且优势科属明显,蹄盖蕨科、鳞毛蕨科、水龙骨科为优势科,卷柏属、铁线蕨属、耳蕨属、瓦韦属、鳞毛蕨属、铁角蕨属为优势属;科的地理分布主要以世界分布为主,占该区总科数的47.6%,属的分布以温带成分为主,占总属数的58.3%,种的地理分布也以温带分布为主,占总种数的48.2%;该区在地理亲缘关系上与河南伏牛山和陕西化龙山最为密切,与四川唐家河、山西五鹿山、北京松山、河北茅荆坝、湖北神农架为其次,与湖南壶瓶山、甘肃祁连山较为疏远,与宁夏南华山的的联系最为疏远。     

POPULATION STRUCTURE AND ESTIMATES OF GENE FLOW IN THE HOMOSPOROUS FERN POLYSTICHUM MUNITUM

Levels and distribution of genetic variation were investigated in the homosporous fern, Polystichum munitum. Homosporous ferns differ from higher vascular plants in that they possess potentially bisexual gametophytes which can produce a completely homozygous sporophyte in a single generation. Because of this, it has long been maintained that ferns possess an inbreeding mating system, resulting in low levels of genetic variation and high levels of homozygosity within populations. The four populations sampled maintain high levels of genetic variation (P? = 0.542; H? = 0.111; ā = 2.23), comparable to that maintained by populations of outcrossing seed plants. The mean fixation index, F, for the four populations was 0.052, indicating no significant deviations from Hardy-Weinberg genotypic expectations. Polystichum munitum distributes most of its genetic variation within rather than among populations. Population-genetic structure was assessed by subdividing each of two large populations into 10 × 10-m subpopulations. Comparisons of genetic variation within and among subpopulations indicated little genetic substructure within either of the artificially subdivided populations. Estimates of interpopulational gene flow (Nm) are extremely high, comparable to those reported for gymnosperms. Statistical estimates of intragametophytic selling are very low, ranging from 0 to 3%. This study suggests that Polystichum munitum is an outcrossing species. Evidence from this and other investigations indicates that fern species do not typically self-fertilize and that mating systems in ferns vary as they do among species of seed plants.