Occasional Outcrossing Research Articles

Extensive sequence duplication in Arabidopsis revealed by pseudo-heterozygosity

BackgroundIt is apparent that genomes harbor much structural variation that is largely undetected for technical reasons. Such variation can cause artifacts when short-read sequencing data are mapped to a reference genome. Spurious SNPs may result from mapping of reads to unrecognized duplicated regions. Calling SNP using the raw reads of the 1001 Arabidopsis Genomes Project we identified 3.3 million (44%) heterozygous SNPs. Given that Arabidopsis thaliana (A. thaliana) is highly selfing, and that extensively heterozygous individuals have been removed, we hypothesize that these SNPs reflected cryptic copy number variation.ResultsThe heterozygosity we observe consists of particular SNPs being heterozygous across individuals in a manner that strongly suggests it reflects shared segregating duplications rather than random tracts of residual heterozygosity due to occasional outcrossing. Focusing on such pseudo-heterozygosity in annotated genes, we use genome-wide association to map the position of the duplicates. We identify 2500 putatively duplicated genes and validate them using de novo genome assemblies from six lines. Specific examples included an annotated gene and nearby transposon that transpose together. We also demonstrate that cryptic structural variation produces highly inaccurate estimates of DNA methylation polymorphism.ConclusionsOur study confirms that most heterozygous SNP calls in A. thaliana are artifacts and suggest that great caution is needed when analyzing SNP data from short-read sequencing. The finding that 10% of annotated genes exhibit copy-number variation, and the realization that neither gene- nor transposon-annotation necessarily tells us what is actually mobile in the genome suggests that future analyses based on independently assembled genomes will be very informative.

Pathotype Complexity and Genetic Characterization of Phytophthora sojae Populations in Illinois, Indiana, Kentucky, and Ohio.

Phytophthora sojae, the causal agent of Phytophthora root and stem rot of soybean, has been managed with single Rps genes since the 1960s but has subsequently adapted to many of these resistance genes, rendering them ineffective. The objective of this study was to examine the pathotype and genetic diversity of P. sojae from soil samples across Illinois, Indiana, Kentucky, and Ohio by assessing which Rps genes were still effective and identifying possible population clusters. There were 218 pathotypes identified from 473 P. sojae isolates with an average of 6.7 out of 15 differential soybean lines exhibiting a susceptible response for each isolate. Genetic characterization of 103 P. sojae isolates from across Illinois, Indiana, Kentucky, and Ohio with 19 simple sequence repeat markers identified 92 multilocus genotypes. There was a moderate level of population differentiation between these four states, with pairwise FST values ranging from 0.026 to 0.246. There were also moderate to high levels of differentiation between fields, with pairwise FST values ranging from 0.071 to 0.537. Additionally, cluster analysis detected the presence of P. sojae population structure across neighboring states. The level of pathotype and genetic diversity, in addition to the identification of population clusters, supports the hypothesis of occasional outcrossing events that allow an increase in diversity and the potential to select for a loss in avirulence to specific resistance genes within regions. The trend of suspected gene flow among neighboring fields is expected to be an ongoing issue with current agricultural practices.

Rare but evolutionarily consequential outcrossing in a highly inbred zoonotic parasite

Recurrent self-mating can result in nearly clonal propagation of biological lineages, but even occasional outcrossing can serve to redistribute variation in future generations, providing cohesion among regional populations. The zoonotic parasite Trichinella spiralis has been suspected to undergo frequent inbreeding, resulting in genetically uniform larval cohorts which differ markedly from one another. Here, we explored the extent of inbreeding for this parasite by determining how genetic variation (at variable microsatellite markers) is distributed among 1379 larvae derived from 41 wild boars in Extremadura, Spain. In particular, we sought to determine how much of the genetic variation in this region’s parasites occurs among the larvae of any given wild boar, and whether each derives from one, or more, parental lineages. We found strong evidence for inbreeding, resulting in genetically distinct parasite subpopulations among the parasites derived from many pairs of wild boar. Fully two-thirds of these parasite cohorts appear to derive from inbred parents; in 10% of the wild boars, parasites were so inbred as to become absolutely fixed in all of the assayed genetic loci. In spite of this, more than one pair of parents appear to have given rise to the infections in one-third of the sampled wild boars, resulting in mixed infections. These mixed infections should slow losses of heterozygosity and multi-locus polymorphism in any given parasite lineage. Such outcrossing should limit distinctions that would otherwise accumulate among transmission chains, thereby enforcing cohesion through the region’s population in spite of its marked departure from panmixia. Conditions of transmission may differ in other regions, where such epidemiological features may engender different evolutionary outcomes.

Fitness consequences of occasional outcrossing in a functionally asexual plant (Oenothera biennis).

Many clonal organisms occasionally outcross, but the long-term consequences of such infrequent events are often unknown. During fiveyears, representing three to five plant generations, we followed 16 experimental field populations of the forb, Oenothera biennis, originally planted with the same 18 original genotypes. Oenothera biennis usually self fertilizes, which, due to its genetic system (permanent translocation heterozygosity), results in seeds that are clones of the maternal plant. However, rare outcrossing produces genetically novel offspring (but without recombination or increased heterozygosity). We sought to understand whether novel genotypes produced through natural outcrossing had greater fecundity or different multigenerational dynamics compared to our original genotypes. We further assessed whether any differences in fitness or abundances through time between original and novel genotypes were exaggerated in the presence vs. absence of insect herbivores. Over the course of the experiment, we genotyped >12,500 plants using microsatellite DNA markers to identify and track the frequency of specific genotypes and estimated fecundity on a subset (>3,000) of plants. The effective outcrossing rate was 7.3% in the first year and ultimately 50% of the plants were of outcrossed origin by the final year of the experiment. Lifetime fruit production per plant was on average 32% higher across all novel genotypes produced via outcrossing compared to the original genotypes, and this fecundity advantage was significantly enhanced in populations lacking herbivores. Among 43 novel genotypes that were abundant enough to phenotype with replication, plants produced nearly 30% more fruits than the average of their specific two parental genotypes, and marginally more fruits (8%) than their most fecund parent. Mean per capita fecundity of novel genotypes predicted their relative frequencies at the end of the experiment. Novel genotypes increased more dramatically in herbivore-present compared to suppressed populations (45% vs. 27% of all plants), countering the increased competition from dandelions (Taraxacum officinale) that resulted from herbivore suppression. Increased interspecific competition likely also lead to the lower realized fitness of novel vs. original genotypes in herbivore-suppressed populations. These results demonstrate that rare outcrossing and the generation of novel genotypes can create high-fecundity progeny, with the biotic environment influencing the dynamical outcome of such advantages.

Population genetics and evolution of the mangrove rivulus Kryptolebias marmoratus, the world's only self-fertilizing hermaphroditic vertebrate.

The mangrove rivulus, Kryptolebias marmoratus (Rivulidae, Cyprinodontiformes), is phylogenetically embedded within a large clade of oviparous (egg laying) and otherwise mostly gonochoristic (separate sex) killifish species in the circumtropical suborder Aplocheiloidei. It is unique in its reproductive mode: K. marmoratus is essentially the world's only vertebrate species known to engage routinely in self-fertilization as part of a mixed-mating strategy of selfing plus occasional outcrossing with gonochoristic males. This unique form of procreation has profound population-genetic and evolutionary-genetic consequences that are the subject of this review.

Environmental and genetic correlates of allocation to sexual reproduction in the circumpolar plant Bistorta vivipara.

• Sexual reproduction often requires more energy and time than clonal reproduction. In marginal arctic conditions, species that can reproduce both sexually and clonally dominate. Plants with this capacity may thrive because they can alter reproduction depending on environmental conditions. Bistorta vivipara is a circumpolar herb that predominately reproduces clonally, but certain environmental conditions promote higher investment in flowers (and possible sexual reproduction). Despite largely reproducing clonally, the herb has high levels of genetic variation, and the processes underlying this paradoxical pattern of variation remain unclear. Here we identified environmental factors associated with sexual investment and examined whether sexual reproduction is associated with higher levels of genetic variation.• We sampled 20 populations of B. vivipara across the high Arctic archipelago of Svalbard. In each population, we measured reproductive traits, environmental variables, and collected samples for genetic analyses. These samples permitted hypotheses to be tested regarding sexual investment and ecological and genetic correlates.• Increased soil nitrogen and organic matter content and decreased elevation were positively associated with investment in flowers. Increased investment in flowers significantly correlated with more genotypes per population. Linkage disequilibrium was consistent with predominant clonality, but several populations showed higher genetic variation and lower differentiation than expected. There was no geographical genetic structure.• In B. vivipara, sexual investment is positively associated with habitat quality. Bistorta vivipara predominantly reproduces clonally, but occasional outcrossing, efficient clonal reproduction, and dispersal by bulbils can explain the considerable genetic variation and weak genetic structure in B. vivipara.

Range-wide genetic analysis reveals limited structure and suggests asexual patterns in the rare forbSenecio macrocarpus

Genetic diversity and recombination underlie the long-term persistence and evolution of species and are strongly influenced by population size, breeding system and plant longevity. Here, we study genetic structure in the rare Senecio macrocarpus in southeastern Australia to guide current conservation practices. Thirteen neutral microsatellite markers and two chloroplast regions were used to survey the 20 known S. macrocarpus populations and one sympatric S. squarrosus population, a morphologically similar species. All markers showed severe excess or deficit of heterozygotes and linkage disequilibrium was significant. Microsatellite markers revealed 100 multi-locus genotypes (MLGs) from 523 S. macrocarpus individuals and a further 4 MLGs from 27 S. squarrosus individuals. MLGs varied in frequency and distribution. At the extremes, one MLG was found 108 times across the sampling region and 66 MLGs were found once. The MLGs of all 38 seedlings genotyped were identical to their seed parents implying an asexual origin. Chloroplast regions showed little variation within S. macrocarpus but differed from S. squarrosus. Chromosome counts for S. macrocarpus revealed the same ploidy level as S. squarrosus (2n = 6x = 60) and pollen–ovule ratios were typical of erechthitoid Senecio species showing self-compatibility. Results suggest that establishment of small populations occur primarily from one extensive source population with indications that both apomixis and selfing may be contributing to its reproduction cycle. We suggest that this species may contribute to future evolutionary processes despite limited genotypic variation and restricted distribution. Its conservation will safeguard evolutionary processes that might occur through occasional outcrossing and hybridization events between sympatric species. © 2015 The Linnean Society of London, Biological Journal of the Linnean Society, 2015, 115, 256–269.

A Genetic Test for Whether Pairs of Hermaphrodites Can Cross-Fertilize in a Selfing Killifish.

Kryptolebias marmoratus, a small killifish that lives in mangrove habitat from southern Florida to Brazil, is one of the planet's only known self-fertilizing hermaphroditic vertebrates. Generation after generation, hermaphroditic individuals simultaneously produce sperm and eggs and internally self-fertilize to produce what are, in effect, highly inbred clones of themselves. Although populations are composed primarily of hermaphrodites, they also contain some true males. The frequency of males in a population varies geographically, from <2% in Florida to as high as 25% in Belize. Males are known to mate occasionally with hermaphrodites, thereby releasing genetic variation that has profound consequences for population genetic structure. However, it is unknown whether hermaphrodites can or do sporadically mate with each other also. Here, we test whether hermaphroditic individuals of the killifish Kryptolebias marmoratus are capable of crossing with one another, in addition to their much more common habits of self-fertilization and occasional outcrossing with pure males. We employ an experimental design in which replicate hermaphrodite pairs were housed together and allowed to reproduce naturally. Among 173 embryos screened at diagnostic microsatellite loci, all were found to result from selfing (i.e., no embryos were the product of a hermaphrodite cross). We thus conclude that hermaphrodite pairs are unlikely to cross, or do so exceedingly rarely.

Genetic structure and diversity ofTriticum monococcumssp.aegilopoidesandT. urartuin Iran

To preliminarily evaluate the genetic diversity of the Iranian diploidTriticumL. gene pool, in this study, a total of 176 individuals belonging toT. monococcumL. ssp.aegilopoides(Link) Thell. andT. urartuThum. ex Gandil. were pre-screened using single-strand conformation polymorphism (SSCP) analysis of theAcc-1andPgk-1loci. A selected set of 76 DNA samples corresponding to the observed SSCP variants were sequenced for both loci and evaluated for nucleotide diversity associated with the taxonomic groups and geographical regions. We found three haplotypes, including one that was new for Iran, at each locus. Population structure and analysis of molecular variation results proved that the collection evaluated could be genetically divided into two distinct groups, which to a great extent was in accordance with the taxonomic classification. A genetic leakage fromT. monococcumssp.aegilopoidesintoT. urartuwas observed during structure analysis, which was inferred on the basis of occasional outcrossing, despite their inbreeding nature. The results revealed that there was no variation within the populations belonging toT. urartu, while a meaningful variation was found between the geographical regions forT. monococcumssp.aegilopoides. The median-joining networks revealed a conflict between morphology and haplotype variation, which was interpreted on the basis of introgressive hybridization, recombination signature and rapid speciation. In conclusion, we suggest that SSCP analysis is a useful tool in regions where thorough sequencing of an enormous number of DNA samples is time consuming and not affordable.

The Ghost of Outcrossing Past in Downy Brome, an Inbreeding Annual Grass

We investigated the frequency of outcrossing in downy brome (Bromus tectorum L.), a cleistogamous weedy annual grass, in both common garden and wild populations, using microsatellite and single nucleotide polymorphic (SNP) markers. In the common garden study, 25 lines with strongly contrasting genotypes were planted in close proximity. We fingerprinted 10 seed progeny from 8 individuals of each line and detected 15 first-generation heterozygotes for a t-value (corrected for cryptic crosses) of 0.0082. Different genotypes were significantly overrepresented as maternal versus paternal parents of heterozygotes, suggesting gender-function-dependent genetic control of outcrossing rates. In 4 wild populations (>300 individuals each), expected heterozygosity ranged from 0.149 to 0.336, whereas t-values ranged from 0.0027 to 0.0133, indicating high levels of both genetic diversity and inbreeding. Up to a third of the individuals in each population belonged to groups with identical or nearly identical SNP genotypes, whereas many of the remaining individuals were members of loose clusters of apparently related plants that probably represent descendants from past outcrossing events. Strict inbreeding in some lineages within a population with occasional outcrossing in others may be related to positive selection on adaptive syndromes associated with specific inbreeding lineages, or possibly to among-lineage differences in genetic regulation of outcrossing.

Systematics and taxonomic delimitation of vegetable, grain and weed amaranths: a morphological and biochemical approach

Amaranthus is a cosmopolitan genus comprising large number of species with many morphotypes cultivars/accessions having diversified morphological features. The species, morphotypes within Amaranthus are very closely related requiring microclassification even revision in present taxonomic status. Most of the works have been done employing grain Amaranthus and their putative ancestors applying morphological and molecular data. In present study along with morphological features, phenolic compounds (secondary metabolites), isozyme profile of acid phosphatase were applied to get comprehensive reflection of interrelationship and relative closeness among the species, morphotypes/accessions of few vegetable, grain and weed Amaranthus. Both the vegetable and grain Amaranthus are supposed to have originated from respective weed progenitor through occasional outcrossing and gradual domestication. The study strongly vouched the previous interpretations regarding interrelationship of grain Amaranthus and their derivation from their putative progenitor also showed clear separate clustering of vegetable and grain Amaranthus with their close relatives. A sharp congruence in cluster pattern was achieved between morphological dendrogram and cumulative dendrogram computed on morphological and biochemical parameters, which is a rare event in systematics.

Fine-scale spatial genetic structure in a predominantly selfing plant: role of seed and pollen dispersal

We present a study of fine-scale spatial genetic structure (SGS) and assess the impact of seed and pollen dispersal on the pattern of genetic diversity in the predominantly selfing Hordeum spontaneum. The study included (1) direct measurement of dispersal in a controlled environment, and (2) analyses of SGS and estimation of the ratio of pollen to seed flow in three natural populations sampled in linear transects at fixed increasing inter-plant distances. Analysis of SGS with 10 nuclear SSRs showed in all three populations a significant autocorrelation for the distance classes of 1 or 2 m and a negative linear relationship between kinship coefficients, calculated for pairs of individuals, and logarithm of geographical distance between members of the pairs. Major seed dispersal (95%) was found to be within 1.2 m from the mother plant. Pollen flow, estimated from the comparison of nuclear and chloroplast variation, was spatially limited as much as was seed dispersal, and tended to be overestimated when measured at spatial scales exceeding that of SGS. We conclude that combined effects of selfing, occasional outcrossing, localized seed dispersal and high plant density create an equilibrium between drift and gene flow in this species resulting in SGS at a very fine spatial scale.

Bromus tectorum (Poaceae) in midcontinental United States: Population genetic analysis of an ongoing invasion

Biological invasions can be substantially influenced by the genetic sampling associated with a species' introduction. As a result, we assessed the genetic and evolutionary consequences of the entry and spread of the invasive grass Bromus tectorum (cheatgrass) across the United States midcontinent through an analysis of 54 populations, using enzyme electrophoresis. On average, these populations display 1.04 alleles per locus (A), 4.1% percent polymorphic loci per population (%P) and an expected mean heterozygosity (H(exp)) value of 0.009. Heterozygotes, which have been rarely reported for B. tectorum in North America, occur in three populations in the midcontinent and are likely novel multilocus genotypes that arose postimmigration. The midcontinent distribution of multilocus genotypes suggests that plant immigrants came directly from either the native range or the eastern United States, or both. Continued dispersal of preadapted genotypes and the assembly of populations that are genetic admixtures may enhance this invasion by increasing both the genetic diversity within populations and the selection of novel genotypes arising from occasional outcrossing. The potential for postimmigration evolution in most species points to the largely unrecognized need to block the introduction of new, potentially aggressive genotypes of an alien species already in the United States.

Population Genetics of Three Important Head Blight Pathogens Fusarium graminearum, F. pseudograminearum and F. culmorum

AbstractHomothallic Fusarium graminearum (teleomorph Gibberella zeae) and anamorphic F. culmorum are destructive pathogens causing Fusarium head blight (FHB) of small‐grain cereals worldwide, while heterothallic F. pseudograminearum (G. coronicola) seems to be restricted to Australia as a FHB pathogen. In a comprehensive treatise of pathogen population genetics, this review summarizes global knowledge of genetic diversity among isolates sampled at various spatial and temporal scales, examines the mechanisms that generate this diversity and explores the implications of pathogen diversity and plasticity to resistance breeding. Despite their different modes of reproduction, there is large variation among isolates of all three species originating from different countries and continents. With a few exceptions, haplotype diversity ranges from 60 to 100% even within populations from individual fields. In F. graminearum, over 90% of the variation is found within populations, even when samples are collected from areas as small as 0.25 m2. Variation among populations is low (4–8%) with negligible population subdivision. This indicates a high level of gene flow (Nm = 8–71) with linkage equilibrium for the majority of selectively neutral molecular marker loci analysed. These findings for F. graminearum point to large random mating populations driven by occasional outcrossing, high gene flow across large geographical distances and a relatively low host‐mediated directional selection. Similar conclusions can be drawn for the Canadian population of F. pseudograminearum, but not for populations from Australia, where different pathogen ecology may have reduced the frequency of sexual recombination. Phylogenetic analyses indicate delineation of lineages in F. graminearum, often along geographically separated lines, while the related F. pseudograminearum is a single recombining species with limited or no lineage development. The anamorphic F. culmorum shows no obvious clonal structure in its population as might have been expected. High levels of diversity within fields may have been caused by balancing selection from frequent alternation between saprophytic and parasitical life cycle and/or a hidden or recently extinct teleomorph. Other mechanisms including parasexual cycles or active transposable elements may also be involved but these have not been investigated as yet. Crosses between and among F. graminearum lineages have shown a rather simple, additive inheritance of pathogenicity and aggressiveness with frequent transgressive segregation in crosses among isolates with moderate aggressiveness. This raises the spectre of highly aggressive and/or toxigenic isolates evolving if a limited range of quantitative trait locus for FHB resistance is deployed on a large scale. Combining more than one genetically distinct sources of resistance, possibly with different modes of action against the pathogen, will be necessary to avoid severe FHB outbreaks in the future.

Introduction history and population genetics of the invasive grass Bromus tectorum (Poaceae) in Canada

The invasive annual Bromus tectorum (cheatgrass) is distributed in Canada primarily south of 52° N latitude in two diffuse ranges separated by the extensive coniferous forest in western Ontario. The grass was likely introduced independently to eastern and western Canada post-1880. We detected regional variation in the grass's genetic diversity using starch gel electrophoresis to analyze genetic diversity at 25 allozyme loci in 60 populations collected across Canada. The Pgm-1a & Pgm-2a multilocus genotype, which occurs in the grass's native range in Eastern Europe, is prevalent in eastern Canada but occurs at low frequency in western Canada. In contrast, the Got-4c multilocus genotype, found in the native range in Central Europe, is widespread in populations from western Canada. Overall genetic diversity of B. tectorum is much higher in eastern Canada than in the eastern U.S., while the genetic diversity in populations in western North America is similar between Canada and the U.S. The distribution of genetic diversity across Canada strongly suggests multiple introduction events. Heterozygous individuals, which are exceedingly rare in B. tectorum, were detected in three Canadian populations. Formation of novel genotypes through occasional outcrossing events could spark adaptive evolution and further range expansion across Canada of this exceedingly damaging grass.

Genetic Diversity and Geographic Differentiation in Tacca chantrieri (Taccaceae): an Autonomous Selfing Plant with Showy Floral Display

Despite considerable investment in elaborate floral displays, Tacca chantrieri populations are predominantly selfing. It is hypothesized that this species might possess considerable spatial or temporal variation in outcrossing rates among populations. To test this hypothesis, genetic variability and genetic differentiation within and among T. chantrieri populations were investigated to find out if they are in agreement with expectations based on a predominantly inbred mating system. Genetic diversity was quantified using inter-simple sequence repeats (ISSR) in 303 individuals from 13 populations taken from known locations of T. chantrieri in China, and from one population in Thailand. Of the 113 primers screened, 24 produced highly reproducible ISSR bands. Using these primers, 160 discernible DNA fragments were generated, of which 145 (90.62 %) were polymorphic. This indicated considerable genetic variation at the species level. However, there were relatively low levels of polymorphism at population levels, with percentages of polymorphic bands (PPB) ranging from 8.75 % to 55 %. A high level of genetic differentiation among populations was detected based on different measures (Nei's genetic diversity analysis: G(ST) = 0.5835; AMOVA analysis: F(ST) = 0.6989). Furthermore, based on levels of genetic differentiation, the 14 populations clustered into two distinct groups separated by the Tanaka Line. High levels of differentiation among populations and low levels of diversity within populations at large spatial scales are consistent with earlier small-scale studies of mating patterns detected by allozymes which showed that T. chantrieri populations are predominantly selfing. However, it appears that T. chantrieri has a mixed-mating system in which self-fertilization predominates, but there is occasional outcrossing. Significant genetic differences between the two distinct regions might be attributed to vicariance along the Tanaka Line. Finally, possible mechanisms of geographic patterns based on genetic differentiation of T. chantrieri are discussed.

Genetic diversity of European populations of the oak fine‐root pathogen Phytophthora quercina

SummaryThe recently discovered oak‐specific fine root plant pathogen Phytophthora quercina is a significant factor in the current phase of European oak decline but its origins and ecology are poorly understood. A genome‐wide analysis of 260 amplified fragment length polymorphism (AFLP) markers was used to examine the genetic diversity of 72 isolates from five oak species at 28 sites in Germany (particularly Bavaria), Italy, France, Hungary and the UK. Within‐site diversity was examined at 16 sites. The limited genetic diversity (within and between sites) and lack of genetic substructuring according to geographic origin or host species suggest the rapid spread of a relatively recently introduced species. Two subgroups were distinguished and these may reflect an initial introduction of isolates of two different genetic backgrounds. The relatively low genetic diversity is probably because of the predominantly inbreeding (homothallic) nature of P. quercina. However, evidence of limited intra‐site diversity, temporal variation and the lack of clonality within the European population suggest that some diversity is being maintained by occasional outcrossing and turnover of a reservoir of long‐lived soil‐borne oospore (sexually derived) inoculum.

Evolutionary history of Caenorhabditis elegans inferred from microsatellites: evidence for spatial and temporal genetic differentiation and the occurrence of outbreeding.

Although diverse biological disciplines employ the nematode Caenorhabditis elegans as a highly efficient laboratory model system, little is known about its natural history. We investigated its evolutionary past using 10 polymorphic trinucleotide and tetranucleotide microsatellites, derived from across the whole genome. These microsatellites were analyzed from the 35 previously available natural isolates from different parts of the world and also 23 new strains isolated from northwest Germany. Our results highlight that C. elegans lineages differentiate genetically with respect to geographic distance and, to a lesser extent, differences in the time of strain isolation. The latter indicates some turnover of strain genotypes at specific locations. Our data also demonstrate the coexistence of highly diverse genotypes in the population from northwest Germany, which is best explained by recent migration events. Furthermore, selfing is confirmed as the primary mode of reproduction for this hermaphroditic nematode in nature. Importantly, we also find evidence for the occurrence of occasional outbreeding. Taken together, these results support the previous notion that C. elegans is a colonizer, whereby selfing may permit rapid dispersal within new habitats even in the absence of potential mates, whereas occasional outcrossing may serve to compensate for the disadvantages of inbreeding. Such information about the natural history of C. elegans should be of great value for an in-depth understanding of the complexity of this organism, including its multifaceted developmental, neurological, or molecular genetic pathways.

The Mating Systems of Some Epiphytic Polypodiaceae

Genetic loads, estimated from sporophyte production by isolated gametophyte cultures, indicate mating systems of intragametophytic selfing in Campyloneurum angustifolium (in part), C. phyllitidis, Phlebodium aureum and Phymatosorus scolopendria, and intergametophytic mating in C. angustifolium (in part), Microgramma heterophylla and Polypodium pellucidum. Polyploidy characterizes the intragametophytic-selfing species, whereas the intergametophytic-mating taxa are diploid. The duplicated loci of polyploid taxa may mitigate the expression of recessive lethal alleles caused by intragametophytic selfing, whereas genetic load probably maintains the mating systems of the intergametophytically mating taxa. Enzyme electrophoretic patterns of fixed heterozygosity support allopolyploid origins of C. phyllitidis and P. aureum and confirm their intragametophytic mating systems. Antheridiogens, present in both groups, may promote intergametophytic mating in diploids through promotion of the early development of male plants in gametophyte populations and bisexuality in isolated gametophytes of polyploids if these gametophytes delay or do not attain insensitivity to their own antheridiogen. In the polyploids, antheridiogens may also alleviate low genetic variability through promotion of occasional outcrossing. The perennial, clone-forming habit of epiphytic Polypodiaceae increases the duration and the physical space occupied by derivatives of a single spore, thus expanding the chance of interaction with a later migrant. Genetic load, duplicated genes, and antheridiogens, together with a perennial and clone-forming gametophyte growth habit, interact to produce successful breeding strategies of these epiphytic species.

An analysis of genetic variation in natural populations of Sticherus flabellatus [R. Br. (St John)] using amplified fragment length polymorphism (AFLP) markers.

Amplified fragment length polymorphisms (AFLPs) were used to characterize the genetic diversity within and among natural populations of Sticherus flabellatus. Eight populations within the Sydney region of New South Wales, Australia were surveyed using 11 primer combinations. A total of 1108 reproducible bands were detected of which 469 (42%) were polymorphic. FST estimates averaged over all polymorphic loci indicated that significant genomic differentiation occurs among populations (average = 0.783). Genetic diversity within populations was assessed according to average heterozygosity (H) and percentage polymorphic loci (P) per population. Within-population diversity ranged from H = 0.12 and P = 33.69 to H = 0.04 and P = 15.99. Analysis of genetic similarity among populations suggested that the eight populations studied fall into two groups of four populations, based on population size and the condition of the habitat. Phenetic analysis (AMOVA) indicated that genetic variation is greater among populations (74.34%) than within populations (25.66%). These findings suggest that the breeding system of S. flabellatus is predominantly inbreeding, with genetic diversity maintained by occasional outcrossing in larger populations. The results presented in this study could provide evidence to support the proposal to protect natural stands of S. flabellatus, which has implications for the Australian horticulture industry.