Ecological genetics of the rotifer Brachionus plicatilis in Soda Lake,Nevada, USA

In an earlier study we found that the rotifer Brachionus plicatilis in Soda Lake, Nevada (USA) maintains extensive genetic variation for three isozymes. This variation was present both within and between different collections. No evidence was found that there was a seasonal succession of distinct electromorphs. In this paper we ask: is the genetic variation maintained by selective neutrality among electromorphs? Parthenogenetic females from ten clones representing seven different composite electromorphs were selected for life table analysis. Substantial main effects of genotype or interactions of genotype with experimental food quantities and temperatures, were found for the intrinsic rates of increase, net reproductive rates, and mean life spans of these clones. We conclude that these fitness differences permit us to reject the hypothesis that genetic variation is maintained because of selective neutrality among clones. Instead, it seems likely that directional selectional may be either too weak or of too brief a duration in Soda Lake for clonal exclusion to occur.     

Behavioural Reproductive Isolation in a Rotifer Hybrid Zone

A hybrid zone between two Brachionus plicatilis rotifer mitochondrial DNA (mtDNA) lineages was recently described in the Iberian Peninsula between a pond (Santed 2) and a lake (Gallocanta). The patterns of mitochondrial and nuclear genetic variation observed suggested that gene flow is mainly male-mediated from the lake to the pond. Here we test two hypotheses: (a) that male-mediated gene flow occurs through assortative mating between individuals from these ponds, (b) that behavioural isolation occurs between the two mtDNA lineages. We isolated, reared and genotyped rotifer clones from resting eggs collected in the sediments of these and two other distant ponds. We devised a quick, inexpensive RFLP method to discriminate between B. plicatilis and its sibling species B. ‘Manjavaeas’ and between both mtDNA B. plicatilis lineages. Behavioural no-choice tests using new-born, virgin males and females were performed between five clones. B. ‘Manjavacas’ and B. plicatilis were reproductively isolated. B. plicatilis clones did not show evidence of reproductive isolation, regardless of their mtDNA lineage, except Santed 2 males, which discriminated strongly against Gallocanta females. These results could help to explain the discrepancies between mitochondrial and nuclear genetic variation reported in the two populations.     

Large Hardy-Weinberg equilibrium deviations in the Daphnia longispina of Lake El Tobar

The population structure of Daphnia longispina in Lake El Tobar, Spain was studied by measuring variation at the aldehyde oxidase (AO), phosphoglucose isomerase (PGI) and phosphoglucose mutase (PGM) loci in each of 1337 individuals from four collections. In 9 of the 12 comparisons between observed allele frequencies and those expected by Hardy-Weinberg equilibrium there was an excess of heterozygotes. We found 27 of the potential number of 54 composite electromorphs (clones) based on the three allozymes. Clone diversities were rather high in all collections. Three clones reached frequencies of over 25% and different clones were dominant in each of the four collections. Strong temporal variation was found in the genetic structure of this Daphnia population. This variation was driven by changes in the relative frequencies of the component clones in the lake rather than by a recruitment of novel clones into the population.We conclude with a consideration of the role of models relating allele and genotype frequencies in populations of cyclical parthenogens. Because the breeding system of these populations infrequently involves recombination between clones, models such as the Hardy-Weinberg have limited value in providing meaningful measures of population structure.     

Ecological genetics of a cyclical parthenogen in temporary habitats

Populations of the rotifer Brachionus plicatilis inhabiting three temporary ponds in Torreblanca Marsh (Castellón, Spain) were regularly screened for allozyme variation, sexual reproduction levels and population densities during an annual cycle. Relevant ecological parameters in the ponds were also recorded. The electrophoretic survey of the three ponds (Poza Sur, Poza Norte and Canal Central) revealed a high level of overall genetic polymorphism in four marker loci, but only 13 multilocus genotypes were found. We classified clones into three clonal groups (SS, SM, L) characterized by unique arrays of alleles in the four marker loci, and significant differences in body shape and size. Clonal group succession took place in Poza Sur; SM clones occurring in spring, SS clones from spring to fall, and L clones from fall to spring. Despite the partial overlapping of sexual periods, the absence of heterozygotes indicates that gene flow is strongly restricted between clonal groups. In the transition periods, the population was far from Hardy—Weinberg equilibrium due to hetereozygote deficiencies in all four markers. In clonal group L, the only polymorphic locus, Pgi, was in Hardy—Weinberg equilibrium. In clonal group SM, both polymorphic loci were in Hardy—Weinberg equilibrium in Poza Norte, but not in Poza Sur. Factor analysis on the limnological parameters recorded during field sampling indicates that genetically different clonal groups are also ecologically specialized. These findings suggest that the taxonomical species B. plicatilis is in fact a species complex. The complete genetic discontinuity model of rotifer population succession is supported by these data. Our results are similar to those found in cladocerans inhabiting temporary habitats.     

Genetic variation in the diatomFragilaria capucina (Fragilariaceae) along a latitudinal gradient across North America

Genetic variation in 126 clones of the planktonic diatom,Fragilaria capucina, collected from seven lakes from Manitoba to Texas, was determined by RAPD analysis. Five primers yielded 48 scorable RAPD fragments in 123 unique combinations. Patterns of genetic variation were analysed by cluster analysis, which showed that most clones grouped together according to the site from which they were collected. This pattern in genetic variation may be a result of the geographically disjunct nature of these populations and/or the different environmental conditions, especially temperature, in the different lakes.     

Differences in demographic parameters among electrophoretic clones of Daphnia obtusa Kurz (Crustacea: Cladocera)

In an 18-month electrophoretic study, a Daphnia obtusa population that recently colonized Lake Orta (a large, severely polluted lake in Northern Italy) showed no genetic variation.Populations from other localities were sampled. From clones established by single females, genotype differences were determined by two life table experiments (using Lake Orta and Lake Maggiore water as a culture medium).As expected, in the Lake Orta water the colonizing clone (LO) had longer survival, produced more newborns and had a larger rate of increase than the other clones. In the Lake Maggiore water the LO clone still fared better than the other clones.     

Phenotypic drift inBradyrhizobium japonicum populations after introduction into soils as established by numerical analysis

The degree of phenotypic variation of the bacterial strains USDA 125-Sp, USDA 138 and USDA 138-SmBradyrhizobium japonicum a long time after introduction was studied in three experimental fields. A total of 54 phenotypic characters were analyzed by constructing a dendrogram based on an hierarchic classification. Strong similarities (92.6, 94 and 95%) were found between the isolates introduced into soil 8, 10 and 13 years ago and between their respectiveB. japonicum parental clones. The dendrogrammic analysis detected a small amount of phenotypic drift, however, between soil isolates and parental clones belonging to the same serogroup (selective effects were found to have generated 0 to 3.9% variation for the USDA 125-Sp inoculum introduced 8 years ago, and 3.2–3.5% after 10 and 13 years, respectively, for the USDA 138 and USDA 138-Sm bacterial inocula) and within the serogroup 125 soil isolates (2.7%). We found a similar evolution of serogroup 125 isolates when compared with parental clones conserved on slant agar at 4°C. When a drift was observed, the isolates from soil presented a lower activity for several enzymes and lower diversity compared with the parental clones.     

A taxonomic approach to evaluation of the charge state model using twelve species of sea anemone

The charge-state model of electrophoretic variation was tested by comparing the distances between nearest electromorphs of five enzyme loci within polymorphic species and among pooled species of sea anemone. If the charge-state model is generally true, and in particular if it allows linear distance between electromorphs to be used as a measure of genetic distance, then electromorphs of different species should be on the same "mobility ladder". Therefore, distances between adjacent electromorphs should be approximately equal for the two sets of comparisons. It was found that distances between adjacent electromorphs for each locus were significantly smaller for the pooled comparisons than within polymorphic species. Thus, it was concluded that much of the variation detected among different species does not conform to the charge-state model, and therefore that distance between electromorphs per se would not be a good measure of genetic distance. However, the charge-state model does appear to adequately account for most of the variation existing as common polymorphisms within species, or between very closely related species. Possible reasons for this apparent difference in the nature of the variation seen within and among species are discussed.     

Molecular Ecology of Rotifers: From Population Differentiation to Speciation

The development of cost-effective molecular tools allowing the amplification of minute amounts of DNA effectively opened the field of molecular ecology for rotifers. Here I review these techniques and the advances they have provided in the understanding of sibling species complexes, clonal structure, resting egg banks, population structure, phylogeographic patterns and phylogenetic relationships in rotifers. Most of the research to date has focused on the rotifer species complex Brachionus plicatilis. The use of DNA sequence and microsatellite variation, in the context of the background knowledge of life history, mating behaviour, and temporal population dynamics in these organisms have revolutionised our views into the processes shaping the genetic diversity in aquatic invertebrates. Rotifers have populations with a very high number of clones in genetic equilibrium. In temporary populations clonal selection is effective in eroding the number of clones. Rotifer populations are strongly differentiated genetically for neutral markers, even at small geographical scales, and exhibit deep phylogeographic structure which might reflect the impact of Pleistocene glaciations. Despite the high potential for dispersal afforded by resting eggs, rotifers display persistent historical colonisation effects, with gene flow effective only at a local scale and with marked isolation by distance. Instances of long-distance transcontinental migration resulting in successful colonisation have also been revealed. B. plicatilis is composed of a group of several ancient species and sympatry is common. Despite this, the presence of cosmopolitan species in this species complex cannot be discounted. I discuss future priorities and point out the main areas where our knowledge is still insufficient.     

An analysis of molecular polymorphism of esterase M produced by motileAeromonas strains

The high levels of electrophoretic polymorphism of esterase M detected in eight distinct hybridization groups of motileAeromonas raise questions of genetic homogeneity of the electromorphs. The 40 electromorphs detected fall in fourM _r classes—75, 80, 90, and 110 kD—and one typical variant belonging to each of these classes was purified. The four purified esterases exhibited the same resistance to heat, topH and to diisopropyl fluorophosphate, the sameK _m values for 1-naphthyl acetate and 1-naphthyl propionate (1mm), and immunological cross-reactions. Within each class, the electromorphs appeared to be related in term of single amino acid substitutions as estimated from their comparative titration patterns. The titration curves of the four purified esterases were strictly parallel suggesting close structural similarities. Thus, despite considerable variation in theirpI,M _F,andM _r values, it seems likely that the variants of esterase M are the products of closely related loci originating from a common ancestral gene.This work was supported by a grant from the Conseil Scientifique de la Faculté Xavier Bichat (Université Paris VII).     

MORPHOLOGICAL AND GENETIC VARIATION IN ECOLOGICALLY CENTRAL AND MARGINAL POPULATIONS OF RUMEX ACETOSELLA L. (POLYGONACEAE)

Six ecologically central (old field) and marginal (strip mine) populations of the hexaploid Rumex acetosella were collected, grown under uniform conditions, and examined for genetic and morphological variation. Extensive electrophoretic variation was found in alcohol dehydrogenase and phosphoglucose isomerase, while other enzymes surveyed showed little or no variation. Hedrick's genotypic measure of identity revealed mean values of 0.506 for central populations, 0.836 for marginal populations, and 0.633 for comparisons of central with marginal populations. Alcohol dehydrogenase phenotypes had significantly fewer electromorphs per individual in marginal populations. Clones of individuals from both environments were subjected to different watering regimes. No significant differences in root/shoot ratio, leaf number, total leaf area, or relative growth rate were found between strip mine and old-field individuals within each watering treatment, although significant differences were found between watering treatments. There are small, but significant amounts of isozyme differentiation between central and marginal populations, while there was no such differentation for morphological characters.     

Variation of Microsatellite Size Homoplasy Across Electromorphs, Loci, and Populations in Three Invertebrate Species

Size homoplasy was analyzed at microsatellite loci by sequencing electromorphs, that is, variants of the same size (base pairs). This study was conducted using five interrupted and/or compound loci in three invertebrate species, the honey bee Apis mellifera, the bumble bee Bombus terrestris, and the freshwater snail Bulinus truncatus. The 15 electromorphs sequenced turned out to hide 31 alleles (i.e., variants identical in sequence). Variation in the amount of size homoplasy was detected among electromorphs and loci. From one to seven alleles were detected per electromorph, and one locus did not show any size homoplasy in both bee species. The amount of size homoplasy was related to the sequencing effort, since the number of alleles was correlated with the number of copies of electromorphs sequenced, but also with the molecular structure of the core sequence at each locus. Size homoplasy within populations was detected only three times, meaning that size homoplasy was detected mostly among populations. We analyzed population structure, estimating F _st and a genetic distance, based on either electromorphs or alleles. Whereas little difference was found in A. mellifera, uncovering size homoplasy led to a more marked population structure in B. terrestris and B. truncatus. We also showed in A. mellifera that the detection of size homoplasy may alter phylogenetic reconstructions. Received: 21 July 1997 / Accepted: 29 January 1998     

Interclonal variation in diel horizontal migration behaviour of the water flea <Emphasis Type="Italic">Daphnia magna</Emphasis>—searching for a signature of adaptive evolution

In shallow temperate lakes, zooplankton populations may exhibit diel horizontal migration (DHM) and move towards macrophytes during the day to avoid fish. Using a natural Daphnia magna population, we undertook an experimental investigation aimed to describe the genetic variation for DHM and to study whether an adaptive micro-evolutionary response occurred to changes in macrophyte coverage and fish predation pressure through time. Twenty-seven D. magna clones were hatched from ephippia in the sediment of shallow Lake Ring, Denmark. This lake was eutrophied during the 20th century and was subject to restoration measures in the 1970s. The DHM behaviour of the clones was observed both in the presence and absence of fish kairomone. Significant interclonal variation in DHM behaviour occurred in both treatments. To study the micro-evolutionary response of the Lake Ring D. magna population, two approaches were used. First, we compared the DHM behaviour of clones derived from ephippia collected at different depths. A comparison was conducted between clones resurrected from the period of eutrophication (1960–1980) and from the period of recovery (1986–2000). A significant treatment (presence and absence of fish kairomone) × period interaction effect was identified, suggesting a significant micro-evolutionary response for DHM behaviour. The D. magna clones exhibited a significantly stronger horizontal migration response during the period of eutrophication than in the recovery phase. Second, clonal means, representing the influence of the genotype on the trait, were correlated with environmental conditions (macrophyte cover, fish predation pressure and Secchi depth). The results of this analysis also suggest that a micro-evolutionary response by Daphnia has occurred in reaction to changes in fish predation pressure. In periods with high fish predation pressure, Daphnia migrated more strongly towards the plants. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

The interplay between colonization history and gene flow in passively dispersing zooplankton: microsatellite analysis of rotifer resting egg banks

Zooplanktonic organisms that disperse passively as diapausing eggs often exhibit surprisingly strong population subdivision given their high colonization ability. Here we attempt to disentangle the impacts of colonization history and gene flow on these organisms by studying the population genetic structure of the rotifer Brachionus plicatilis. The resting egg banks of B. plicatilis in fourteen salt lake populations in the Iberian Peninsula were examined using seven microsatellite loci. A remarkably high degree of geographical structuring was found (F_st=0.43), with a significant pattern of isolation by distance. Microsatellite loci were in genetic equilibrium, ruling out inbreeding as an important force in population structuring. Comparisons are drawn with previously published phylogeographical data. Surprisingly, introgression of nuclear genes was detected in neighbouring populations with divergent mtDNA haplotypes. These results stress the long lasting impact of colonization history and the modulating effect of gene flow at local scales in these organisms.     

Patterns of genetic diversity in Great Lakes bloaters (Coregonus hoyi) with a view to future reintroduction in Lake Ontario

The originally diverse ciscoe fish fauna of the Laurentian Great Lakes has suffered many extinctions and local extirpations. Bloaters (Coregonus hoyi) are presumed extirpated from Lake Ontario and the reintroduction of this deepwater fish is under consideration. Given the demographic fluctuations of this species in the other Great Lakes and its recent intralacustrine origin, we sought to identify a genetically diverse and similar source of C. hoyi via an analysis of genetic diversity and population structure using 10 microsatellite loci. Despite well-documented demographic declines, we found no genetic evidence of bottlenecks in 12 C. hoyi samples from the four potential donor lakes (Huron, Michigan, Superior and Nipigon). By contrast, evidence of bottlenecks in historical samples of C. artedi from Lake Ontario suggested that standard genetic methods frequently used to identify population bottlenecks can only detect very severe and long-lasting demographic declines in naturally large populations. Patterns of genetic differentiation and assignment tests indicated that C. hoyi from Lake Huron and Lake Michigan, which are not differentiated, are genetically most similar to Lake Ontario ciscoes. The small available sample of deepwater ciscoes recently caught in Lake Ontario did not allow determining if these represent a small undetected C. hoyi population or a recent invasion of the deep section by C. artedi. On the basis of genetic criteria, we conclude that C. hoyi from any location within Lake Huron or Lake Michigan would be judicious sources of breeders for reintroducing C. hoyi in Lake Ontario.     

Spatial Variation in <Emphasis Type="Italic">Streptomyces</Emphasis> Genetic Composition and Diversity in a Prairie Soil

Understanding how microbial genotypes are arrayed in space is crucial for identifying local factors that may influence the spatial distribution of genetic diversity. In this study we investigated variation in 16S rDNA sequences and rep-PCR fingerprints of Streptomyces stains isolated from prairie soil among three locations and four soil depths. Substantial variation in Streptomyces OTU (operational taxonomic unit) and BOX-PCR fingerprint diversity was found among locations within a limited spatial area (1 m²). Further, phylogenetic lineages at each location were distinct. However, there was little variation in genetic diversity among isolates from different soil depths and similar phylogenetic lineages were found at each depth. Some clones were found at a localized scale while other clones had a relatively widespread distribution. There was poor correspondence between 16S rDNA groupings and rep-PCR fingerprint groupings. The finding of distinct phylogenetic lineages and the variation in spatial distribution of clones suggests that selection pressures may vary over the soil landscape.     

Studies on life history characteristics of <Emphasis Type="Italic">Brachionus plicatilis</Emphasis> O. F. Müller (Rotifera) in relation to temperature,salinity and food algae

Effects of temperature (18, 24, and 30°C), salinity (5–40 ppt, five intervals) and algal foods (Synechococcus sp., Chlorella pyrenoidosa, Isochrysis zhanjiangensis, Dunaliella salina and Tetraselmis cordiformis) on the life table demography of six geographical Brachionus plicatilis sensu stricto clones, which had been identified according to the prevalent taxonomy and biometric analysis of B. plicatilis sensu lato, were studied. The results showed that temperature, salinity and temperature × salinity significantly influenced the life history parameters. Genotype (clone) had no effect on the population growth rate but did influence the net reproductive rate, generation time and lifespan. All rotifer clones showed the expected increase in population growth rate with increasing temperature. B. plicatilis s. s. attained a higher population growth rate at low–medium salinities (5–20 ppt) than at high salinities (25–40 ppt). The equivalent spherical diameter (ESD) of food algae, salinity and ESD × salinity had significant effects on the life history parameters. In this case, genotype had no effect on population growth rate, net reproductive rate and generation time but did influence lifespan. The population growth rate of B. plicatilis s. s. evaluated against particle retention spectrum of algae at two salinities resulted in bell-shaped curves. Dunaliella salina with an ESD = 7.7 μm was considered to be the best food for B. plicatilis s. s. while Synechococcus appeared to be an inadequate food algae.     

HERITABILITY OF MANTLE AREOLAR CHARACTERS IN AULACOSEIRA SUBARCTICA (BACILLARIOPHYTA)1

Are size and arrangement of valve mantle areolae in Aulacoseira Thwaites adapted to light intensity? To test one criterion demonstrating an adaptation, heritability experiments were run on isolates of Aulacoseira subarctica (Müller) Haworth. Several clones of A. subarctica were isolated from Yellowstone Lake (Wyoming, USA), Lewis Lake (Wyoming), and East Rosebud Lake (Montana, USA). Two to four clones from each lake were grown in batch cultures under three irradiance levels: 2, 11.4, and 115 μmol photons·m^?2·s^?1. Five randomly chosen valves for each of two replicates of each clone were examined using a scanning electron microscope for a total of 300 valves. Size measurements were taken for each valve examined, and images of mantle areolae were captured on film at a magnification of 20,000×. Each image was digitized, and quantitative morphometric areolar characters were measured. A quantitative genetic analysis was performed within each light environment for the mean area of the external opening of mantle areolae, the mean distance between areolae within pervalvar striae, and the mean distance between pervalvar striae. Resulting estimates of heritability from among‐lake and within‐lake analyses indicate that all three mantle areolar characters could presently respond to selection and thus have potentially done so in the past.     

Effects of population outcrossing on rotifer fitness

Background  

Outcrossing between populations can exert either positive or negative effects on offspring fitness. Cyclically parthenogenetic rotifers, like other continental zooplankters, show high genetic differentiation despite their high potential for passive dispersal. Within this context, the effects of outcrossing may be relevant in modulating gene flow between populations through selection for or against interpopulation hybrids. Nevertheless, these effects remain practically unexplored in rotifers. Here, the consequences of outcrossing on the rotifer Brachionus plicatilis were investigated. Cross-mating experiments were performed between a reference population and three alternative populations that differed in their genetic distance with regard to the former. Two offspring generations were obtained: F1 and BC ('backcross'). Fitness of the outcrossed offspring was compared with fitness of the offspring of the reference population for both generations and for three different between-population combinations. Four fitness components were measured throughout the rotifer life cycle: the diapausing egg-hatching proportion, clone viability (for the clones originating from diapausing eggs), initial net growth rate R for each viable clone, and the proportion of male-producing clones. Additionally, both the parental fertilisation proportion and a compound fitness measure, integrating the complete life cycle, were estimated.     

Molecular genetics of rotifers: preliminary restriction napping of the mitochondrial genome of Brachionus plicatilis

Methods are presented to extract and purify mitochondrial DNA from the rotifer Brachionus plicatilis. The mtDNA obtained is of sufficient purity for digestion with restriction endonucleases. EcoR I restriction patterns are presented for 4 geographically separated clones. A restriction map based on digestion with 5 different restriction enzymes is included for one of these clones. Finally, use of mtDNA analysis for studies on the population structure and biogeography of rotifers is discussed.