A SYNOPSIS OF THE GENUS CHARA,SERIES GYMNOBASALIA (SUBSECTION WILLDENOWIA RDW)

The series Gymnobasalia, a sharply differentiated subgeneric complex within the genus Chara, consists of at least seven morphologically distinct species. All seven species are reproductively isolated from one another. No evidence of even limited hybridization between the seven was encountered. The two dioecious members of the complex occupy restricted and, possibly partially overlapping ranges in South America. Monoecious members are in general more widely distributed but still appear to have originated in Central or South America. The holotype of C. zeylanica Klein ex Willdenow was examined and found to be tetrascutate. For the abundant and widely distributed monoecious-conjoined, octoscutate member of the complex, previously shown to be distinct from C. zeylanica, the earlier name C. haitensis Turpin is here validated. Chara foliolosa Muhl. ex Willdenow is considered conspecific with, but to hold priority over, C. sejuncta Braun.     

TAXONOMY OF CHARA BRAUNII: AN EXPERIMENTAL APPROACH1

Results from the present study suggest that Chara braunii encompasses a complex of geographical races or incipient species that although morphologically similar, are isolated to varying degrees ranging from partial interfertility to complete reproductive incompatibility. Most of the 6 clones studied were interfertile to the extent that fertilization was followed by the development of black oospores, but subsequent germination tests disclosed that many of these either failed to germinate or gave rise to offspring that were partially or completely sterile. No support was encountered for the suggestion that C. braunii is of recent development or that it has been distributed rapidly throughout the world.     

Two in one: cryptic species discovered in biological control agent populations using molecular data and crossbreeding experiments

There are many examples of cryptic species that have been identified through DNA‐barcoding or other genetic techniques. There are, however, very few confirmations of cryptic species being reproductively isolated. This study presents one of the few cases of cryptic species that has been confirmed to be reproductively isolated and therefore true species according to the biological species concept. The cryptic species are of special interest because they were discovered within biological control agent populations. Two geographically isolated populations of Eccritotarsus catarinensis (Carvalho) [Hemiptera: Miridae], a biological control agent for the invasive aquatic macrophyte, water hyacinth, Eichhornia crassipes (Mart.) Solms [Pontederiaceae], in South Africa, were sampled from the native range of the species in South America. Morphological characteristics indicated that both populations were the same species according to the current taxonomy, but subsequent DNA analysis and breeding experiments revealed that the two populations are reproductively isolated. Crossbreeding experiments resulted in very few hybrid offspring when individuals were forced to interbreed with individuals of the other population, and no hybrid offspring were recorded when a choice of mate from either population was offered. The data indicate that the two populations are cryptic species that are reproductively incompatible. Subtle but reliable diagnostic characteristics were then identified to distinguish between the two species which would have been considered intraspecific variation without the data from the genetics and interbreeding experiments. These findings suggest that all consignments of biological control agents from allopatric populations should be screened for cryptic species using genetic techniques and that the importation of multiple consignments of the same species for biological control should be conducted with caution.     

MOLECULAR AND REPRODUCTIVE CHARACTERIZATION OF SIBLING SPECIES IN THE EUROPEAN EARWIG (FORFICULA AURICULARIA)

The European earwig (Forficula auricularia) was formerly thought to present a mosaic of populations differing in their reproductive biology. We show that it is comprised of two as yet unrecognized sibling species. The molecular divergence between the two species, for a 627-bp amplified fragment overlapping the COI and COII mitochondrial loci, is six times larger than intraspecific variation. A species with two clutches a year lives predominantly in lowland and oceanic European habitats. A species with one clutch a year—except in the Mediterranean area where it has two clutches—lives predominantly in highland and continental European habitats. They both invaded North America during the 20th century, respectively, from the west and the east coasts, with no apparent mixing of their populations. The two species can occur in sympatry in Europe and are reproductively isolated by nearly complete failure to produce F₁ hybrids.     

REPRODUCTIVE ISOLATION BETWEEN SYMPATRIC RACES OF PEA APHIDS. I. GENE FLOW RESTRICTION AND HABITAT CHOICE

Determining the extent and causes of barriers to gene flow between genetically divergent populations or races of single species is an important complement to post facto analyses of the causes of reproductive isolation between recognized species. Sympatric populations of pea aphids (Acyrthosiphon pisum Harris, Homoptera: Aphididae) on alfalfa and red clover are highly genetically divergent and locally adapted. Here, hierarchical estimates of population structure based on F_st suggest that gene exchange between closely adjacent aphid populations on the two hosts is highly restricted relative to that among fields of the same host plant. Although these host-associated races are presently considered to be the same subspecies, they appear to be significantly reproductively isolated, suggesting incipient speciation. Habitat (host) choice was investigated as the first in a temporal series of factors that could reduce gene exchange between these sympatric populations. Field studies of winged colonists to newly planted fields of each host suggest pronounced habitat fidelity. This result was verified using replicated observations of the host choice behavior of different aphid genotypes for which the relative demographic performance on each host was known. These laboratory observations of behavior revealed a strong genetic correlation between habitat choice (or acceptance) and the relative performance in each habitat. Because mating occurs on the host plant, habitat choice in this system leads to assortative mating and is therefore a major cause of reproductive isolation between the sympatric pea aphid populations on alfalfa and clover. However, the extent of dispersal between hosts estimated from the field study of winged colonists (9–11%) is too great to be consistent with the genetic divergence estimated between the races. This suggests that barriers to gene flow other than host choice also exist, such as selection against migrants or hybrids in the parental environments, hybrid sterility, or hybrid breakdown.     

SYSTEMATICS AND POPULATION GENETICS OF FIRE ANTS (SOLENOPSIS SAEVISSIMA COMPLEX) FROM ARGENTINA

Specimens of seven fire ant species collected from their native ranges in Argentina were studied by protein electrophoresis and morphological analysis. Concordance between the genetic and morphological character sets is strong (96% agreement on identifications), suggesting that recognition of reproductively isolated populations and partitioning of intra- and interspecific variation can in most cases be achieved using appropriate characters of either type in this taxonomically difficult group. Genetic differentiation between native (Argentina) and introduced (USA) conspecific populations of two species, Solenopsis invicta and S. richteri, is rather typical of the differentiation existing between conspecific populations found within either country. Furthermore, there appears to have been little reduction of variability (heterozygosity) at enzyme loci following colonization by either species of the United States, although some rare alleles have been lost in the introduced populations. Hybridization is rare between S. invicta and S. richteri where their native ranges overlap in central Argentina, in contrast to the extensive hybridization of these species in the United States, suggesting that prezygotic barriers to gene flow have been compromised in introduced populations. Phylogenetic analysis of the seven species indicates that S. invicta and S. richteri are relatively distantly related within the S. saevissima complex. Given that hybrids between these species in the United States suffer little apparent loss of fitness, genomic incompatibilities generally may be insufficient to create effective postzygotic barriers to interspecific gene flow in this group of ants.     

Genetic analysis of sympatric migratory ecotypes of Arctic charr Salvelinus alpinus: alternative mating tactics or reproductively isolated strategies?

Three populations of Arctic charr Salvelinus alpinus from southern Baffin Island were previously identified to display variable migratory phenotypes, with an anadromous component of the population and another remaining resident in fresh water. In this study, 14 microsatellite markers were used to help distinguish between two alternative hypotheses to explain the co‐existence of the two ecotypes: that the two ecotypes originate from a single population and are the result of a conditional mating tactic or that the migratory ecotypes are reproductively isolated populations utilizing alternative migratory strategies. In two of the three replicate systems, F_ST values between the resident and anadromous individuals were non‐significant, while they were significant in a third sampling location. Bayesian clustering analysis implemented in structure, however, failed to identify any within‐location clustering in all three sampling locations. It is concluded from these analyses that the life‐history ecotypes are most likely conditional mating tactics, rather than reproductively isolated populations. Other evidence in favour of the alternative mating tactic hypothesis is briefly reviewed, and implications for management of those populations are discussed.     

THE GEOGRAPHIC PATTERN OF GENETIC DIFFERENTIATION BETWEEN HOST ASSOCIATED POPULATIONS OF RHAGOLETIS POMONELLA (DIPTERA: TEPHRITIDAE) IN THE EASTERN UNITED STATES AND CANADA

True fruit flies in the Rhagoletis pomonella sibling species group are at the center of a long-standing debate concerning modes of speciation. The allopatric separation of populations is widely thought to be a prerequisite for speciation in sexually reproducing animals. However, speciation in the R. pomonella group appears to have occurred sympatrically as a consequence of these flies shifting and adapting to new host plants. The sympatric shift of R. pomonella from its native host hawthorn to introduced domestic apple, which occurred approximately 150 years ago, provides a test of whether host specialization is sufficient to allow populations to differentiate in the absence of geographic barriers to gene flow. We report the results of a geographic study of allozyme variation for hawthorn and apple infesting populations of R. pomonella across the eastern United States and Canada. Six loci consistently show significant allele frequency differences at paired apple and hawthorn sites. These six loci map to three different regions of the genome, and linkage disequilibrium exists between non-allelic genes within each of these regions. Allele frequencies for five of the six loci displaying host associated differences also co-vary significantly with latitude. Inter-host divergence is, therefore, superimposed on north-south clinal patterns of intra-host variation such that the magnitude of genetic divergence between hawthorn and apple flies is a function of latitude. The findings suggest that partially reproductively isolated “host races” can evolve in sympatry as a consequence of R. pomonella infesting new host plants. Host recognition and host associated developmental traits are discussed as important factors differentiating apple and hawthorn flies.     

LOCAL VARIATION IN THE GENETIC BASIS OF PAEDOMORPHOSIS IN THE SALAMANDER AMBYSTOMA TALPOIDEUM

The hypothesis that local isolated populations differed in the genetic basis for life-history traits was tested in the salamander Ambystoma talpoideum. Genetic basis was defined as the specific genetic architecture (additive and nonadditive) that contributes, along with maternal and environmental factors, to the phenotype. All crosses within and between three populations were made to produce nine F₁ populations. Nine within-population crosses produced the F₂ generation. This design does not permit an estimation of the exact nature of the genetic basis (e.g., additive, nonadditive) for any trait within populations. However, hybrid dissimilarity in the F₂ generation was taken as evidence of a different genetic basis for a trait in each population. The genetic basis of life-history pathway (metamorphosis vs. paedomorphosis) and per capita fecundity differed between two populations. The genetic basis of life-history pathway, per capita fecundity, survival, and growth rate was similar between the remaining sets of populations. This study and related ones (Semlitsch and Wilbur, 1989; Semlitsch et al., 1990) suggest that a heterochronic shift that causes rapid morphological evolution between metamorphosis and paedomorphosis (a macroevolutionary pattern) can evolve independently and does not require a macromutation or other nonmicroevolutionary mechanisms.     

GENETIC AND MORPHOLOGICAL IDENTIFICATION OF FUCUS RADICANS SP. NOV. (FUCALES,PHAEOPHYCEAE) IN THE BRACKISH BALTIC SEA1

Brown seaweeds of the genus Fucus occupy a wide variety of temperate coastal habitats. The genus is evolutionary dynamic with recent radiations to form morphologically distinct taxa. In the brackish Baltic Sea, fucoids are the only perennial canopy‐forming macroalgae. The most northern populations of Fucus occur permanently submerged in extremely low salinity (3–5 psu). These are currently referred to as Fucus vesiculosus L. but are morphologically distinct with a narrow frond without bladders. We report here that a population of this unique morphotype is reproductively isolated from a truly sympatric population of common F. vesiculosus and conclude that the northern morphotype represents a previously undescribed species. We describe Fucus radicans sp. nov., which is attached and dioecious with broadly elliptic receptacles, characterized by a richly branched narrow flat frond (2–5 mm), short thallus (<26 cm), and a high capacity for vegetative recruitment of attached plants. Analysis of five highly polymorphic microsatellite DNA loci showed genetic differentiation between sympatric populations of F. radicans and F. vesiculosus, whereas allopatric populations of the same species revealed a coherent pattern of genetic variation. Sequences of the RUBISCO region in F. radicans were identical to or differing at only one to two dinucleotide positions from those of F. vesiculosus, indicating a recent common origin of the two species.     

GENETIC POPULATION STRUCTURE OF PSEUDO‐NITZSCHIA PUNGENS (BACILLARIOPHYCEAE) FROM THE PACIFIC NORTHWEST AND THE NORTH SEA1

Several species of the diatom Pseudo‐nitzschia produce the neurotoxin domoic acid (DA). Consumption of fish and shellfish that have accumulated this potent excitotoxin has resulted in severe illness and even death in humans, marine mammals, and seabirds. Pseudo‐nitzschia pungens (Grunow ex Cleve) Hasle is a cosmopolitan diatom commonly occurring in the waters of the Pacific Northwest (PNW) and the eastern North Atlantic, including the North Sea. However, genetic and physiological relationships among populations throughout this large geographic distribution have not been assessed. Population genetic parameters (e.g., Hardy–Weinberg equilibrium, linkage equilibrium, F_ST) calculated for P. pungens collected from the Juan de Fuca eddy region in the PNW indicated the presence of two distinct groups that were more divergent from each other than either was from a P. pungens sample from the North Sea. Geographic heterogeneity was also detected within each of the two PNW groups. These results suggested that the populations of P. pungens recently mixed in the Juan de Fuca eddy region (a seasonally retentive feature off the coasts of Washington State, USA, and Vancouver Island, Canada) but did not exchange genetic material by sexual reproduction. Alternatively, these two groups may be cryptic (morphologically identical, but reproductively isolated) species. Identifying cryptic diversity in Pseudo‐nitzschia is important for bloom prediction and aiding the identification of molecular markers that can be used for rapid detection assay development.     

THE GEOGRAPHICAL DISTRIBUTION OF B-CHROMOSOMES OF XANTHISMA TEXANUM DC. (ASTERACEAE). I. SURVEY OF THE RANGE

The distribution of B-chromosomes throughout the range of Xanthisma texanum has been determined from approximately 750 individuals from 80 wild populations in Texas, Oklahoma, and New Mexico. Counts of meiotic chromosome figures of n = 4_II + 1 B_I and n = 5_II (4_II + IB_II) are reported for the first time for subsp. texanum var. texanum and for subsp. texanum var. orientalis. The wide occurrence of B-chromosomes in natural populations of subsp. drummondii is also documented. B-chromosomes are present in 22 (27.2%) randomly distributed populations whose characteristics are in no way distinct from populations without B-chromosomes. In populations with B-chromosomes present, 19.8% of the plants contained one or two B-chromosomes, but in total only 5.8 % of all plants contained one or two B-chromosomes. All populations in one isolated population group of subsp. drummondii lack B-chromosomes.     

EVOLUTIONARY STUDIES OF ATRIPLEX: A RELIC GIGAS DIPLOID POPULATION OF ATRIPLEX CANESCENS

Growing on shifting sand dunes in central Utah is a small endemic population of a gigas form of A triplex canescens. Whereas normal A. canescens usually grows to a height of three to four feet and occasionally to five or six feet, the gigas form often reaches ten and sometimes twelve feet. All normal A. canescens so far examined (67 populations) have 2n = 36 chromosomes; the gigas form has 2n = 18 chromosomes. Several lines of evidence suggest that the gigas form is a relic diploid and the normal form is an autotetraploid derived from it. The growth rate of seedlings and new twigs is nearly twice as great in the diploid as in the tetraploid. Seed germination is faster and much better in the diploid. The tetraploid is reproductively isolated from the diploid because of a much earlier flowering period. The diploid plants possess many attributes which make them uniquely adapted to the drifting sand dune habitat.     

BIOCHEMICAL TAXONOMY OF MARINE PHYTOPLANKTON BY ELECTROPHORESIS OF ENZYMES. I. THE CENTRIC DIATOMS THALASSIOSIRA PSEUDONANA AND T. FLUVIATILIS1,2

Diatom systematics depends almost entirely upon structure of the silica shell. It is not known to what extent the taxonomic species, as defined by shell structure, corresponds to the genetic species—i.e., to the reproductively isolated population. As an approach to this problem, we report here a comparison of enzymes by electrophoresis. We have examined the genetic constitution of a number of clones of (presumably) the same species for each of 2 closely related, centric diatom species: Thalassiosira pseudonana Hasle and Heimdal and T. fluviatilis Hustedt. The 4 clones of T. fluviatilis form a distinct group, clearly separated from all the T. pseudonana clones. Within T. pseudonana, 4 estuarine clones and one reef clone form a group that is distinctly different from 4 oceanic clones. A single clone of T. pseudonana from the Continental Slope waters is intermediate between these 2 groups and probably shares genes with both groups, indicating that the 2 T. pseudonana groups are not genetically isolated. We conclude that i) within groups, isolates are closely related even though they originated from different continents; and, ii) T. pseudonana is subdivided into ecological races.     

Analysis of cross-compatibility inEriastrum densifolium (Polemoniaceae)

Cross-compatibility within and among populations of the perennialEriastrum densifolium was investigated to determine if sterility barriers have evolved within this polytypic species, and to correlate any such barriers to previously known patterns of genetic and morphological variation. Furthermore, the reproductive affinities of the endangeredE. densifolium subsp.sanctorum are investigated. Results indicate that no single population or group of populations is reproductively isolated from all other populations, and that F1 hybrids are fully fertile. Interspecific crosses (withE. eremicum andE. sapphirinum) were largely unsuccessful. The endangeredE. densifolium subsp.sanctorum demonstrates high interfertility with all other populations.     

REPRODUCTIVE CHARACTER DISPLACEMENT OF EPICUTICULAR COMPOUNDS AND THEIR CONTRIBUTION TO MATE CHOICE IN DROSOPHILA SUBQUINARIA AND DROSOPHILA RECENS

Interactions between species can alter selection on sexual displays used in mate choice within species. Here we study the epicuticular pheromones of two Drosophila species that overlap partially in geographic range and are incompletely reproductively isolated. Drosophila subquinaria shows a pattern of reproductive character displacement against Drosophila recens, and partial behavioral isolation between conspecific sympatric versus allopatric populations, whereas D. recens shows no such variation in mate choice. First, using manipulative perfuming experiments, we show that females use pheromones as signals for mate discrimination both between species and among populations of D. subquinaria. Second, we show that patterns of variation in epicuticular compounds, both across populations and between species, are consistent with those previously shown for mating probabilities: pheromone compositions differ between populations of D. subquinaria that are allopatric versus sympatric with D. recens, but are similar across populations of D. recens regardless of overlap with D. subquinaria. We also identify differences in pheromone composition among allopatric regions of D. subquinaria. In sum, our results suggest that epicuticular compounds are key signals used by females during mate recognition, and that these traits have diverged among D. subquinaria populations in response to reinforcing selection generated by the presence of D. recens.     

THE EVOLUTIONARY HISTORY OF PARTHENOGENETIC CNEMIDOPHORUS LEMNISCATUS (SAURIA,TEIIDAE). I. EVIDENCE FOR A HYBRID ORIGIN

Chromosomes and allozymes were studied from chromosomally distinct unisexual (races B and C) and bisexual (races D and E) populations of the teiid lizard Cnemidophorus lemniscatus, and from selected outgroup taxa (C. murinus, C. nigricolor, Ameiva ameiva, and A. auberi). Karyotyping confirmed the racial identity of individuals and showed that the chromosomal composition of populations at specific localities has remained the same for 20 years. All individuals of both unisexual populations were heterozygous for a pericentric inversion that distinguishes D and E bisexuals. Also, the unisexuals were all heterozygous for 8 of 11 protein loci for which D and E were fixed or nearly fixed for different alleles. Most of these alleles represent derived states relative to the other Cnemidophorus and Ameiva analyzed, and the fixed heterozygote condition at these nine markers provides unequivocal support for the hypothesis that the unisexual C. lemniscatus arose by hybridization between ancestors genetically similar to extant D and E populations. At the remaining three loci for which D and E show fixed differences, the unisexuals were homozygous rather than heterozygous. This suggests that either (1) allozymes have been lost by mutations to null, silent, or convergent mobility states, (2) ancestral genotypes were similar to but not identical with the extant D and E races, and/or (3) limited recombination may occur between unisexual genomes. Allozyme-based genetic distances between D and E were large, suggesting that bisexual races D and E are genetically isolated; each race should be accorded full species status. This conclusion is supported by the absence of any clear biochemical evidence for their monophyly with respect to the other Cnemidophorus examined. Cladistic analyses of 17 phylogenetically informative loci revealed two equally parsimonious shortest trees, one supporting monophyly and the other paraphyly of the C. lemniscatus complex. Further testing of the monophyly of C. lemniscatus requires additional data. With the present study, the evidence that all parthenogenetic Cnemidophorus are of hybrid origin is complete.     

DIFFERENTIATION AND INTEGRATION OF THE GENOME IN POPULATIONS OF THE MARINE COPEPOD TIGRIOPUS CALIFORNICUS

Geographically separated populations of the intertidal copepod Tigriopus californicus are sharply differentiated at several enzyme-encoding gene loci. Two studies were performed to investigate the extent to which the gene pools of local populations are organized into harmoniously interacting (or “coadapted”) gene complexes. In the first, the effects of interpopulation hybridization on development time were assessed. Results showed that while F₁ hybrids did not differ from parental lines, mean F₂ developmental times were as much as 50% longer. The second study used two unlinked enzyme polymorphisms as genetic markers to determine the genotypic specificity of F₂ hybrid breakdown. For two sets of parental populations, the relative viabilities of the different two-locus genotypes were determined from segregation ratios among the F₂ progeny. Sharp deviations from Mendelian ratios were observed; in the extreme, a block of genes marked by the Me^F allozyme from the LJ (La Jolla) population was found to be nearly lethal when homozygous in the F₂ of LJ × AB (Los Angeles) crosses. This same block of genes had a tenfold higher viability in crosses between LJ and SC (Santa Cruz). In the AB × LJ crosses, the two marker loci had independent (multiplicative) effects on viability. In the SC × LJ crosses, deviations from the multiplicative model were observed; the data indicate that parental homozygous genotypes have higher viability than predicted by independence, while nonparental homozygotes have lower than predicted viability. These results suggest that substantial integration of the genome has occurred within natural T. californicus populations.     

Production of Oogonia and Oospores of <Emphasis Type="Italic">Leptolegnia chapmanii</Emphasis> Seymour (Straminipila: Peronosporomycetes) in <Emphasis Type="Italic">Aedes aegypti</Emphasis> (L.) Larvae at Different Temperatures

The aquatic oomycete fungus Leptolegnia chapmanii Seymour is pathogenic to mosquito larvae, but it has been little studied since it was first isolated. Although studies have been performed on different biological isolates of L. chapmanii around the world, they were made on zoospores and a very little or even nothing is known about the sexual stage (oogonia and oospores), which allows L. chapmanii to remain in the environment when conditions are not favorable. The main objective of this study was to determine the relationship between temperature and time of onset of L. chapmanii oogonia and oospores in Ae. aegypti larvae. Leptolegnia chapmanii-infected IV instar Ae. aegypti larvae were incubated at different temperatures between 5 and 45°C and photoperiod-controlled for 90 days. The number of oogonia and oospores was examined daily for each tested temperature. As was expected, low temperatures extended the times of oogonia formation, as much as seven times. Likewise, temperatures significantly affect the number of oogonia produced.