Variation in therbcL sequence ofStegnogramma pozoi subsp.mollissima (Thelypteridaceae) in Japan

Stegnogramma pozoi (Lagasca) K. Iwats. subsp.mollissima (Fischer ex Kunze) K. Iwats. is one of the most common ferns in Japan with a very broad geographical distribution and habitat ranges. We examined the intrasubspecific variation of therbcL gene, and found three types in the subspecies (Atami-1, Atami-2, and Tateyama types). The Atami-1 and 2 types shared three apomorphic nucleotides and their geographical distribution is intermingled. Thus, we considered them to have intraspecific variation. In contrast, the Tateyama type had 4–5 different nucleotides than the Atami types, and showed a more northern distribution. Leaf morphology of the Tateyama type is distinct from that of the Atami types, though the leaf shape in Tateyama type varies considerably and in some cases overlaps with other types. Ferns have fewer morphological features that are systematically significant than flowering plants because they lack complex reproductive organ. It is reasonable to expect that species recognition of ferns based only on leaf morphology can be fine tuned by molecular features. Variation in therbcL sequences might be useful as an indication of significant biological units.     

Genetic variation in populations of the morphologically and ecologically variable fern <Emphasis Type="Italic">Stegnogramma pozoi</Emphasis> subsp. <Emphasis Type="Italic">mollissima</Emphasis> (Thelypteridaceae) in Japan

In Japanese Stegnogramma pozoi subsp. mollissima (Fisher ex Kunze) K. Iwats. there is the intrasubspecific variation among rbcL sequences. Northern and southern plants are genetically differentiated for maternally inherited cpDNA. In the present study we examined allozyme polymorphisms to test the hypothesis that northern and southern plants may be separate species. Based on allozyme data, the degree of gene flow among populations was estimated to be large. The artificial crossing experiments between cpDNA haplotypes also suggested that isolation has not developed among these cpDNA haplotypes. However, interpopulation genetic differentiation in cpDNA was observed even in the small area at the foot of Mt. Hakone, and the cpDNA haplotypes appear to have different habitat preferences. Received: May 1, 2001 / Accepted: September 30, 2001     

Origin and taxonomic affinities of Thelypteris (subgen. Stegnogramma) burksiorum (Thelypteridaceae)

This paper examines the ecology, frond and spore morphology, gametophytes, and molecular biology ofThelypteris burksiorum to reveal its taxonomic affinities and phytogeographic origin. The species is endemic to Winston County in northern Alabama (U.S.A.) and only occurs in cave-like rockhouse habitats. A related species,T. pilosa, is widespread in central and southern Mexico, Guatemala, and Honduras.Thelypteris pilosa is extremely variable and our data suggest that current recognition of the varietiesmajor andpilosa is not warranted.Thelypteris pilosa is clearly closely related and likely sister toT. burksiorum. Thelypteris burksiorum exhibits the unusual production of gametophytic propagules. This, in combination with current levels of genetic diversity and reproductive biology, suggests thatT. burksiorum is an ancient Tertiary relict and not an example of recent long distance dispersal. Selection in rockhouse habitats has driven species away from their normal life cycle to partial or complete reliance on the gametophyte generation. The combination of a reduced yet fertile sporophyte and a gametophyte that may rely on gametophytic proliferation placesT. burksiorum in an intermediate evolutionary position relative to other rockhouse pteridophytes.     

Genetic variation in small,isolated fern populations

Abstract. Differences in genetic variability of several small, isolated populations of four fern species in a restricted area in the Swiss lowlands reflect differences in breeding system, population size, the degree of population fragmentation, and ecological requirements. The investigated populations of Asplenium septentrionale show only little genetic variability (isozyme variation) without gene flow among populations (based on the banding pattern of multi-locus phenotypes), and they persist for long periods despite the small population sizes. In Asplenium ruta-muraria, genetic variability is correlated with age. Young populations show no genetic variation, while old populations show some. All individuals of Polypodium vulgare investigated, either epiphytic or epilithic, share exactly the same enzyme phenotype. The results for these three species can be related to predominance of inbreeding, lack of inbreeding depression, polyploidy, long-distance dispersal, production of large amounts of diaspores, single-spore colonization, and perennial life cycles. Genetic variability in these three species does not seem to be absolutely necessary for the maintenance of their populations. Ecological and demographic factors are considered to be more important. An isolated, glacial relict population of diploid Asplenium viride shows high variability in two out of eight enzyme systems, which may be due to prevailing outbreeding. We discuss aspects of the importance of genetics and life history for conservation biology.     

Genetic variation of Casuarina equisetifolia subsp equisetifolia and C. equisetifolia subsp incana populations on the northern coast of Senegal

The genetic variation of 70 individual samples of Casuarina equisetifolia (L. Johnson) subsp equisetifolia and C. equisetifolia subsp incana growing along the northern coast of Senegal was analyzed with RAPD markers. Of the 160 primers tested, five were chosen; they generated 1396 reproducible bands and 61 polymorphic bands that were scored. This result showed a narrow genetic variation among (4.36%) and within (5.90%) C. equisetifolia subsp equisetifolia and C. equisetifolia subsp incana plantation sites. The genetic variation at each site revealed a high degree of polymorphism in Potou (5.90%) and low diversity in Retba (3.06%). In the dendrogram analyses, each sampling site was formed by two main groups. Similar results were found for the dendrograms based on the RAPD data gathered from the five different sites. These dendrograms revealed several polytomies in one of the subgroups, suggesting replication of the same specimens in different sites along the Senegalese coast. The RAPD data support the hypothesis that these populations are of the same provenance, subject to hybridization and inbreeding depression.     

Two types of partial fertility in a diploid population of the fern Thelypteris decursive-pinnata (Thelypteridaceae)

Two types of abnormal sporophytes were observed in a population of diploid Thelypteris decursive-pinnata. Most sporophytes in this population exhibited regular chromosome pairing, resulting in the formation of 30 bivalents in meiosis I; however, they produced abortive spores to various degrees. Some formed large globose spores at low frequencies, most likely to be unreduced diplospores. The other type of abnormal sporophyte underwent synaptic failure to form 60 univalents at meiosis I, but produced fertile spores, mostly large globose ones at low frequencies. The globose spores were considered unreduced diplospores because the gametophytes arising from them produced tetraploid sporophytes by gametophytic selfing. One tetraploid formed only univalents at meiosis I. Allozyme variation was not detected in this population, although neighboring ordinary diploid populations exhibited it to a certain degree. The sympatric occurrence and allozyme uniformity of the two groups suggest that both are offspring of a founder sporophyte, which may have possessed two types of mutated recessive genes responsible for the spore sterility and the synaptic failure in meiosis. Unreduced spores formed by these two types may play an important role in the polyploid speciation of this species.     

Genetic diversity in Dactylorhiza majalis subsp. majalis populations (Orchidaceae) of northern Poland

We analysed 16 populations of Dactylorhiza majalis subsp. majalis from northern Poland, simultaneously utilizing both morphological and molecular data. Genetic differentiation was examined using five microsatellite loci, and morphological variation was assessed for 23 characters. At the species level, our results showed a moderate level of genetic diversity (A = 6.00; A_e = 1.86; H_o = 0.387; F_IS = 0.139) which varied between the studied populations (A = 2.60–4.20; A_e = 1.68–2.39; H_o = 0.270–0.523; F_IS = ?0.064–0.355). A significant excess of homozygotes was detected in five population, while excess of heterozygotes was observed in four populations, but the latter values were statistically insignificant. Moderate, but clear between population genetic differentiation was found (F_ST = 0.101; p < 0.001). Considering pairwise‐F_ST and number of migrants among populations, we recognized three population groups (I, II, III), where the first could be further divided into two subgroups (Ia, Ib). These three groups differed with respect to gene flow values (N_m = 0.39–1.12). The highest number of migrants per generation was noticed among populations of subgroup Ia (8.58), indicative of a central panmictic population with free gene flow surrounded by peripatric local populations (Ib) with more limited gene flow. Geographic isolation, habitat fragmentation and limited seed dispersal are inferred to have caused limitations to gene flow among the three indicated population groups. There was a significant correlation between the morphological and genetic distance matrices. A weak but significant pattern of isolation by distance was also observed (r = 0.351; p < 0.05).     

Genetic variation is not concordant with morphological variation in the fern Asplenium hookerianum sensu lato (Aspleniaceae)

Species status cannot be adequately determined when partitions are based on only a single morphological character. For instance, the sympatry of plants with broad and narrow pinnules in the fern Asplenium hookerianum sensu lato from New Zealand creates the impression that two entities are present. The narrow-pinnuled plants are sometimes segregated as a distinct species, A. colensoi. However, this variation in pinnule morphology could equally be infraspecific, and only additional data can resolve this uncertainty. Analyses using AFLP DNA-fingerprinting and DNA sequencing of the chloroplast trnL-trnF region indicate that genetic variation in A. hookerianum sensu lato is not concordant with pinnule morphology. Consequently, the recognition of A. colensoi is not supported.     

Genetic variation and its relationship to population size in reintroduced populations of pink sand verbena, Abronia umbellata subsp. breviflora (Nyctaginaceae)

Genetic monitoring of reintroduced plantpopulations can allow assessment of the successin establishing new populations thatgenetically resemble native populations. Weused a PCR-based method (Intersimple SequenceRepeats) to quantify genetic variation in fourreintroduced populations of Abroniaumbellata ssp. breviflora, an annualforb native to the Pacific Coast that isstate-listed endangered in Oregon. Thereintroduced populations ranged in size from 18to 4,111 individuals in the year they weresampled. Genetic variation was also quantifiedin the natural population that served as theseed source for the reintroduction efforts. Atotal of 77 loci (bands) was observed using twoISSR primers, providing 65 polymorphic loci. Asignificant, positive regression was observedbetween the log of population size for the fivepopulations and genetic variation when measuredas percent polymorphic loci (P), expectedheterozygosity (H_e> ), and with adissimilarity index (1 – S_xy) based on bandsharing. Two of the reintroduced populationsmaintained approximately 90% of the geneticvariation we observed in the source population. Based on these results, we predict thatreintroduced populations of A. u. ssp.breviflora that have at least 1,000individuals should maintain 90% of the geneticvariation of the source population.     

Genetic distinctness of three widespread and morphologically variable species of Drupella (Gastropoda,Muricidae)

Corallivorous gastropods of the genus Drupella have caused considerable damage to corals at widely separated reefs in the Indo-Pacific. Morphological variability of Drupella species within and between areas has caused taxonomic confusion. To clarify the relationships, we examined allozyme variation at 16 gene loci in samples from Western Australia, Queensland and Japan. Within sites, the species D. cornus, D. rugosa and D. fragum were distinguishable individually by each of 9 to 11 loci, with average genetic identities of about 0.25. The differences extended across sites, whereas the conspecific genetic identities over distances up to 6000 km were 0.86 to 1.00, supporting the view that there are three widespread species of Drupella. Nevertheless, there is much variation within species for allozymes, size, shape and colour.     

A global phylogeny of Stegnogramma ferns (Thelypteridaceae): generic and sectional revision,historical biogeography and evolution of leaf architecture

The thelypteroid fern genus Stegnogramma s.l. contains around 18–35 species and has a global, cross-continental distribution ranging from tropical to temperate regions. Several genera and infrageneric sections have been recognized previously in Stegnogramma s.l., but their phylogenetic relationships are still unclear. In this study, we present a global phylogeny of Stegnogramma s.l. with the most comprehensive sampling to date and aim to pinpoint the phylogenetic positions of biogeographically and taxonomically important taxa. Based on the reconstructed historical biogeography and character evolution, we propose a new (infra)generic classification and discuss the diversification of Stegnogramma s.l. in a biogeographical context. New names or combinations are made for 12 (infra)species, including transferring the monotypic species of Craspedosorus to Leptogramma. Finally, we discuss a possible link between leaf architecture and ecological adaptation, and hypothesize that the increase in leaf dissection and free-vein proportion is an adaptive feature to cool climates in Stegnogramma s.l.     

Genetic and phenotypic variation across a hybrid zone between ecologically divergent tree squirrels (Tamiasciurus)

A hybrid zone along an environmental gradient should contain a clinal pattern of genetic and phenotypic variation. This occurs because divergent selection in the two parental habitats is typically strong enough to overcome the homogenizing effects of gene flow across the environmental transition. We studied hybridization between two parapatric tree squirrels (Tamiasciurus spp.) across a forest gradient over which the two species vary in coloration, cranial morphology and body size. We sampled 397 individuals at 29 locations across a 600‐km transect to seek genetic evidence for hybridization; upon confirming hybridization, we examined levels of genetic admixture in relation to maintenance of phenotypic divergence despite potentially homogenizing gene flow. Applying population assignment analyses to microsatellite data, we found that Tamiasciurus douglasii and T. hudsonicus form two distinct genetic clusters but also hybridize, mostly within transitional forest habitat. Overall, based on this nuclear analysis, 48% of the specimens were characterized as T. douglasii, 9% as hybrids and 43% as T. hudsonicus. Hybrids appeared to be reproductively viable, as evidenced by the presence of later‐generation hybrid genotypes. Observed clines in ecologically important phenotypic traits—fur coloration and cranial morphology—were sharper than the cline of putatively neutral mtDNA, which suggests that divergent selection may maintain phenotypic distinctiveness. The relatively recent divergence of these two species (probably late Pleistocene), apparent lack of prezygotic isolating mechanisms and geographic coincidence of cline centres for both genetic and phenotypic variation suggest that environmental factors play a large role in maintaining the distinctiveness of these two species across the hybrid zone.     

Genetic variation of esterase D (EsD) polymorphism in Indian populations

Data are presented on the phenotypes and gene frequencies of esterase D (EsD) polymorphism in various endogamous caste groups of Patiala and Faridkot districts of Punjab, north-west India. The frequency of the EsD2 allele in these groups varies from a minimum of 0.157 in Khatris to a maximum of 0.253 in Ramdasia Sikhs, a range quite typical of the north Indian populations. Examination of the available data on esterase D system in indigenous populations demonstrates the existence of a north-south cline in the distribution of the EsD2 allele in India.     

Genetic variation within and among populations of Fomitopsis pinicola (Basidiomycetes)

RAPD polymorphisms were used to reveal the genetic population structure of Fomitopsis pinicola from 5 populations in southwestern Sweden with outliers in Finland and Norway. Eleven primers were used on 35 isolates. Using the Analysis of Molecular Variance (AMOVA), the total variance was divided into 3 hierarchical components: 84% within populations, 11% among regions, and 5% among populations within regions. The 3 large Swedish populations contained 95% of the variation within them. The statistical significance of these patterns was supported by permutation tests. The similarity index between all genets ranged from 0.10 to 0.72, with an average of 0.45. Genets from the same population could form more than one cluster in the neighbor-joining analysis. Some of these clusters were also supported by parsimony jack-knifing. This pattern is tentatively explained by establishment of spores from different basidiomata. The result of somatic incompatibility tests and RAPD markers were compared and this comparison indicated that compatible reactions do not necessarily imply genetic identity. Sampling from cut sections of infected trees revealed that multiple infections were present in a single tree and that the fungus probably infects host-trees by basidiospores, arriving via the air. Each somatically incompatible genet characteristically monopolized only part of a resource unit. Spore trapping showed no evidence of long distance spore dispersal, but this is probably due to the limited experiment, since the genetic analysis suggested a high rate of gene flow.     

Genetic diversity and gene flow in the morphologically variable, rare endemics Begonia dregei and Begonia homonyma (Begoniaceae)

Begonia dregei and B. homonyma (Begoniaceae), rare plants endemic to coastal forests of eastern South Africa, are two closely related species with high levels of variation among populations in the shape of leaves. Distribution of genetic variation and genetic relatedness were investigated in 12 populations of B. dregei and seven of B. homonyma using polyacrylamide gel electrophoresis. Twelve of the 15 enzyme loci examined were polymorphic, but only seven loci were polymorphic within at least one population. Genetic diversity measures indicated that the among-population gene differentiation represents >90% of the total genetic component in both species considered individually or combined. This indicated restricted gene flow, consistent with the limited dispersal abilities of Begonia generally and the ancient separation of isolated forest patches. Genetic distances among populations are much higher than usually found within species. Allozyme data provide no support for the recognition of B. dregei and B. homonyma as distinct species.     

Genetic variation of European grayling (Thymallus thymallus) populations in the Western Balkans

In order to elucidate genetic composition of European grayling (Thymallus thymallus) populations in the Western Balkans, the partial mitochondrial DNA (mtDNA) control region was sequenced and 12 microsatellite loci genotyped in 14 populations originating from tributaries of the Adriatic and Danube drainages. Eleven mtDNA haplotypes were found, one confined to the Adriatic clade, one to the Alpine group and the rest to the ‘Balkan’ grayling phylogenetic clade. Haplotypes from the Balkan clade were confined to the Danube drainage and constituted two groups: northern group with haplotypes found in the Slovenian part of the Danube drainage, and southern group, consisting from Bosnia–Herzegovina and Montenegro. Substantial genetic distance between northern and southern groups of haplotypes (0.75–1.8%) and well supported divisions within the northern group indicate very structured grayling population within the studied Danube basin that most probably did not evolve due to vicariance but rather as a consequence of multiple colonization waves that might have occurred during the Pleistocene. Furthermore, genetic distance of ~4% between Adriatic and Danube populations’ haplotypes, suggest that their separation occurred in mid-Pliocene. These findings imply a complex colonization pattern of the Western Balkans drainages. Microsatellite data also confirm high genetic diversity in Western Balkans populations of grayling (on average 7.5 alleles per microsatellite locus and H _exp 0.58). Limited stocking activities were detected based on microsatellites and mtDNA data. Regarding current knowledge of grayling phylogeography appropriate management strategies were proposed to preserve unique, autochthonous grayling populations in Western Balkan.     

Genetic variation and population structure in Scandinavian wolverine (Gulo gulo) populations

Wolverine (Gulo gulo) numbers in Scandinavia were significantly reduced during the early part of the century as a result of predator removal programmes and hunting. Protective legislation in both Sweden and Norway in the 1960s and 1970s has now resulted in increased wolverine densities in Scandinavia. We report here the development of 15 polymorphic microsatellite markers in wolverine and their use to examine the population sub-structure and genetic variability in free-ranging Scandinavian wolverine populations as well as in a sample of individuals collected before 1970. Significant subdivision between extant populations was discovered, in particular for the small and isolated population of southern Norway, which represents a recent recolonization. Overall genetic variability was found to be lower than previously reported for other mustelids, with only two to five alleles per locus and observed heterozygosities (H(O)) ranging from 0.269 to 0.376 across the examined populations, being lowest in southern Norway. Analysis of the mitochondrial DNA control region revealed no variation throughout the surveyed populations. As the historical sample did not show higher levels of genetic variability, our results are consistent with a reduction in the genetic variation in Scandinavian wolverines that pre-dates the demographic bottleneck observed during the last century. The observed subdivision between populations calls for management caution when issuing harvest quotas, especially for the geographically isolated south Norwegian population.     

Genetic differentiation among populations of Sesleria albicans Kit. ex Schultes (Poaceae) from ecologically different habitats in central Europe

As observed for many other plant species, the populations of Sesleria albicans in Central Europe are located in habitats, which differ to a high degree from each other with regard to ecological factors such as nutrients, light and water as well as in type of land use. The species colonizes limestone cliffs, pavements, screes, grazed and mown grasslands, heaths, fens and open woodlands. In this study, we used random amplified polymorphic DNA (RAPD) analysis to investigate the genetic differentiation among 25 populations of S. albicans from six different types of habitat (beech forests, alpine and lowland rocky ridges, lowland screes, fens, calcareous grasslands). With RAPD analysis, 344 fragments could be amplified, of which 95.9% were polymorphic. The level of polymorphism ranged from 29.7 to 56.7% polymorphic bands per population and was correlated with population size. In an analysis of molecular variance (AMOVA), used to detect variation among individuals within populations, among populations from the same habitat and among different habitats, most of the genetic variation was found within populations (62.06%) and among populations from the same habitat (33.36%). In contrast, only a very low level of differentiation could be observed among different habitats (4.58%). The results of our study give only little evidence for an ecotypic differentiation of Sesleria albicans. This differentiation is principally conceivable, but obviously not related to the investigated RAPD loci.     

Genetic variation and population structure in Korean populations of eurya japonica (Theaceae)

We investigated levels of genetic diversity, population genetic structure, and gene flow in Eurya japonica, a widespread and broad-leaved evergreen dioecious tree native to Japan, China, Taiwan, and the southern and southwestern coast of the Korean Peninsula. Starch-gel electrophoresis was conducted on leaves collected from 1,000 plants in 20 Korean populations. All 12 loci examined were polymorphic in at least one population, and the mean number of alleles per locus was 3.79. In addition, mean observed population heterozygosity (Ho_p = 0.425), expected heterozygosity (He_p = 0.462), and total genetic diversity (H_T = 0.496) were substantially higher than average values for species with similar life history traits. Although significant differences in allele frequency were detected between populations at all loci (P < 0.001), <7% of the genetic variation was found among populations (F_ST = 0.069). There was a significant negative correlation between genetic identity and distance between populations (r = -0.341; P < 0.05), but this explained only a small amount of the diversity among populations. Indirect estimates of the number of migrants per generation (Nm) (3.37, calculated from F_ST; 3.74, calculated from the mean frequency of eight private alleles) indicate that gene flow is extensive among Korean populations of E. japonica. Factors contributing to the high levels of genetic diversity found within populations of E. japonica include large and contiguous populations, obligating outcrossing (dioecious plant), high fecundity, and long generation time. Occasional seed dispersal by humans and pollen movement by domesticated honey bees may further enhance gene flow within the species.