Genetic insights into the biogeography of the southeastern North American endemic, Ceratiola ericoides (Empetraceae)

The southeastern United States harbors an unusually large number of endemic plant taxa, which may reflect the refugial nature of the region during Pleistocene glacial maxima. Understanding the genetic diversity and structure of extant plant taxa can provide insights into the biogeographical processes that shaped them genetically. Here, we investigate the levels and partitioning of allozyme diversity in the southeastern North American endemic, Ceratiola ericoides, which displayed greater genetic variation and structure than other endemics. Central Florida populations represent a center of genetic diversity, whereas South Carolina and Georgia Fall Line sandhill populations have a subset of the Central Florida genetic diversity and may be relicts of a once continuous distribution. This much broader, continuous distribution throughout the southeastern United States occurred during glacial maxima when the scrub habitat, dominated by C. ericoides, expanded considerably owing to drier climatic conditions. Georgia Coastal Plain populations appear to have been independently founded more recently by propagules from Central Florida and the Fall Line sandhills because they have an even more limited subset of genetic diversity and greater genetic heterogeneity among populations. Since their establishment, coastal plain populations appear to have had little, if any, gene exchange among each other or with the relatively proximate Fall Line sandhill populations. These data underscore the importance of understanding the genetic composition and historical biogeography of species before intelligent management or restoration decisions can be made regarding their preservation.     

Sex ratio and spatial pattern of males and females in the dioecious sandhill shrub, Ceratiola ericoides ericoides (Empetraceae) Michx

Sex ratio, size, age, and spatial pattern were investigated for males and females of the dioecious shrub Ceratiola ericoides ericoides (Empetraceae) Michx. within seven mapped populations in Georgia and South Carolina, USA. Among the sites studied, two are regularly burned and one site long-unburned. Age was estimated from node counts of individual shrubs. Only one (fire-suppressed) population showed a female-biased sex ratio, while all others did not differ significantly from 1:1. Mean age estimates did not differ between sexes at any site nor did mean shrub canopy diameter. Bivariate Ripley’s K analysis with a null hypothesis of random labeling was used to investigate whether any of the mapped populations exhibited spatial segregation of the sexes (SSS). No population showed strong evidence of SSS. Rather all sites but one showed males and females to be associated (though not significantly) at a scale of 1–10 m. At a scale of 10–35 m male and female shrubs were located randomly with respect to one another at all sites.     

Allelopathic effects and root distribution of Ceratiola ericoides (Empetraceae) on seven rosemary scrub species

We studied the root distribution and the effects of leachates from the dominant shrub in rosemary scrub, Florida rosemary (Ceratiola ericoides), on germination of seven subordinate rosemary scrub species. For rosemary scrub specialists, (Eryngium cuneifolium and Hypericum cumulicola), germination was suppressed by the leaf and litter leachates. For species that are not found exclusively in rosemary scrub (Liatris ohlingerae, Polygonella basiramia, Paronychia chartacea, and Palofoxia feayi) litter and leaf leachate did not suppress germination significantly. Species limited to gaps in rosemary scrub (E. cuneifolium, H. cumulicola, and Lechea deckertii) showed reduced germination from rosemary leachates while species not limited to rosemary-free gaps (L. ohlingerae and P. feayi) were not affected by rosemary leachates. Rosemary root abundance was greatest near shrubs, at a shallow depth, and at sites not recently burned. As rosemary scrub patches age, rosemary roots are more likely to interact with herbaceous species in gaps.     

Local genetic structure in a North American epiphytic lichen, Ramalina menziesii (Ramalinaceae)

Epiphytic lichens possess unique life history traits that can have conflicting effects on genetic structure: symbiotic mutualism between a fungus with its algal or cyanobacterial photobiont, association with a host plant, and ability to reproduce sexually and asexually. Our study species, Ramalina menziesii, has small ascospores that can facilitate long-distance gene movement, and it is capable of clonal reproduction. The goals of this study are to test whether different haplotypes were differentially distributed across host plant species, to look for evidence of asexual vs. sexual reproduction, and to assess the local genetic structure of the population. We sampled individuals from multiple trees of three oak species in four lichen subpopulations within a savanna ecosystem. Using DNA sequence data from four fungal nuclear loci, we found no tendency for host specialization. Alleles were randomly distributed across subpopulations. The frequency of multilocus genotypes was consistent with a randomly mating population. Sexual reproduction involving relichenization appeared to be the predominant mode of reproduction of R. menziesii at this study site. We found no significant local genetic structure suggesting widespread gene flow at the local scale. The genetic structure of this lichen is comparable to that of widely distributed epiphytic plants.     

Origin of the Hawaiian endemic mints within North American Stachys (Lamiaceae)

The Hawaiian endemic mints constitute a major island radiation, displaying a remarkable diversity of floral, fruit, and vegetative features. Haplostachys and Phyllostegia have flowers associated with insect pollination, whereas Stenogyne has flowers typical of bird pollination. The three genera had been thought to be closely related to East Asian members of Lamioideae tribe Prasieae because of the fleshy nutlets borne by Phyllostegia and Stenogyne. We evaluated the origins of the Hawaiian mints using phylogenetic analyses of DNA sequence data from the plastid rbcL and trnL intron loci and the nuclear ribosomal 5S nontranscribed spacer. The Hawaiian genera were found to be monophyletic but deeply nested inside another lamioid genus, Stachys. In particular, they were found to be most closely related to a group of temperate North American Stachys from the Pacific coast, suggesting that the Hawaiian mints derived from a single colonization event from western North America to the Hawaiian Islands. Furthermore, Stachys, which contains amphiatlantic and transberingian clades, was found to be polyphyletic, with some species more closely related to Gomphostemma, Phlomidoschema, Prasium, and Sideritis than to other species of Stachys. Based on chromosomal evidence and our phylogenetic analyses, we hypothesize that the Hawaiian mints may be polyploid hybrids whose reticulate genomes predate the Hawaiian dispersal event and are derived from Stachys lineages with flowers exhibiting insect- vs. bird-pollination characteristics. Thus, the Hawaiian endemic mints may provide yet another insular system for the combined study of polyploidy, hybrid cladogenesis, and adaptive radiation.     

Spatial genetic structure of Aquilegia taxa endemic to the island of Sardinia

Background and Aims

The Mediterranean Basin is one of the most important regions for the Earth''s plant biodiversity; however, the scarcity of studies on fine scale patterns of genetic variation in this region is striking. Here, an assessment is made of the spatial genetic structure of all known locations of the three Sardinian endemic species of Aquilegia in order to determine the relative roles of gene flow and genetic drift as underlying evolutionary forces canalizing the divergence of Sardinian Aquilegia taxa, and to see if the spatial genetic structure found fits the current taxonomic differentiation of these taxa.

Methods

DNA from 89 individuals from all known locations of Aquilegia across Sardinia was analysed by means of amplified fragment length polymorphism (AFLP) markers. Both principal co-ordinates analysis (PCoA) and Bayesian clustering analyses were used to determine the spatial genetic structure irrespective of any taxonomic affiliation. Historical effects of gene flow and genetic drift were assessed by checking for the existence of isolation-by-distance patterns.

Key Results

STRUCTURE and PCoA analyses revealed a pattern of genetic variation geographically structured into four spatial genetic groups. No migration–drift equilibrium was detected for Aquilegia in Sardinia, when analysed either as a whole or in individual groups. The scenario approached a Case III pattern sensu Hutchinson and Templeton, which is associated with extreme isolation conditions where genetic drift has historically played a dominant role over gene flow.

Conclusions

The pattern of genetic variation of Sardinian taxa of Aquilegia indicates that genetic drift has been historically more influential than gene flow on population structure of Sardinian species of Aquilegia. Limited seed dispersal and divergent selection imposed by habitat conditions have been probably the main causes reinforcing post-Pleistocene geographical isolation of Aquilegia populations. The spatial genetic structure found here is not fully compatible with current taxonomic affiliations of Sardinian Aquilegia taxa. This is probably a consequence of the uncoupling between morphological and genetic patterns of differentiation frequently found in recently radiated taxa.     

Population genetic structure of an endangered Utah endemic, Astragalus ampullarioides (Fabaceae)

The endangered Shivwits milkvetch, Astragalus ampullarioides, is a perennial, herbaceous plant. This Utah endemic was federally listed as endangered in 2001 because of its high habitat specificity and low numbers of individuals and populations. All habitat currently occupied by A. ampullarioides was designated as critical by the U.S. Fish and Wildlife Service in 2006 as a result of conservation litigation. We used AFLP markers to assess genetic differentiation among the seven extant populations and quantified genetic diversity in each. Six different AFLP markers resulted in 217 unambiguous polymorphic loci. We used multiple methods to examine any changes in population genetic structure in this species over time. Results indicate that A. ampullarioides had much higher gene flow among populations in the past, but has since fragmented into regional genetic units. These regions further fragmented genetically, and extant populations have differentiated through genetic drift. Populations had low levels of gene flow, even between geographically close populations. Rapid urban development reduces gene flow among regions and encroaches on populations of A. ampullarioides and remaining patches of unoccupied habitat. The genetic makeup of each of the extant populations should be carefully considered in management decisions such as population establishment or augmentation.     

Matriarchal genetic population structure of North American beluga whales Delphinapterus leucas (Cetacea: Monodontidae)

The North American beluga whale Delphinapterus leucas population has been divided into a number of putative geographical stocks based upon migration routes and areas of summer concentration. Nucleotide sequences of the mitochondrial DNA (mtDNA) control region were used to assess whether these geographical stocks are genetically distinct. Beluga whale samples from 25 sites were collected primarily from aboriginal subsistence hunts across North America from 1984 to 1994. Thirty-nine mtDNA haplotypes were identified in 628 beluga samples. No differences were found in the distribution of haplotypes between male and female beluga whales at any sampling site. These haplotypes segregated into two distinct assemblages in both a haplotype network and a neighbour-joining tree. The haplotype assemblages had a geographically disjunct distribution that suggests postglacial recolonization of the North American Arctic from two different refugia.
An analysis of molecular variance based on haplotype relationships and frequency indicated genetic heterogeneity among beluga whale summering groups ( P ≤ 0.001). Sequence divergence estimates between sampling sites also indicated geographical differentiation, particularly between samples taken at east Hudson Bay or St Lawrence River and the western or central Arctic. The results of this study show a high degree of philopatry to specific summering areas by this highly mobile animal.     

The cryptic genetic structure of the North American captive gorilla population

Western lowland gorillas (Gorilla gorilla gorilla) were imported from across their geographical range to North American zoos from the late 1800s through 1974. The majority of these gorillas were imported with little or no information regarding their original provenance and no information on their genetic relatedness. Here, we analyze 32 microsatellite loci in 144 individuals using a Bayesian clustering method to delineate clusters of individuals among a sample of founders of the captive North American zoo gorilla collection. We infer that the majority of North American zoo founders sampled are distributed into two distinct clusters, and that some individuals are of admixed ancestry. This new information regarding the existence of ancestral genetic population structure in the North American zoo population lays the groundwork for enhanced efforts to conserve the evolutionary units of the western lowland gorilla gene pool. Our data also show that the genetic diversity estimates in the founder population were comparable to those in wild gorilla populations (Mondika and Cross River), and that pairwise relatedness among the founders is no different from that expected for a random mating population. However, the relatively high level of relatedness (R = 0.54) we discovered in a pair of known breeding pairs reveals the need for incorporating genetic relatedness estimates in the captive management of western lowland gorillas.     

Spatial and genetic structure within populations of wild American ginseng (Panax quinquefolius L., Araliaceae)

Spatial structure and fine-scale genetic structure were analyzed for the medicinal plant American ginseng (Panax quinquefolius L.) to more fully understand biological processes within wild populations. P. quinquefolius has been harvested for more than 250 years and is now considered threatened or rare throughout its range. Plants within four protected and four unprotected populations were significantly clumped based on Ripley's univariate analysis. Analysis with Ripley's bivariate test determined that juvenile plants were significantly clumped with adult plants at the shortest distance classes in all populations. Although plants were highly clumped, we found that significant fine-scale genetic structure was restricted to the shortest distance classes based on estimates of coancestry (f(ij)). In most cases, estimates of f(ij) were more significant among juveniles than among adults, especially at the shortest distance classes. The spatial structure of ginseng seems to result from the establishment and persistence of plants in favorable microhabitats coupled with limited seed dispersal around maternal individuals. There were no differences in patterns of fine-scale genetic structure between protected and unprotected populations.     

Achene structure and taxonomy of North American Bolboschoenus (Cyperaceae)

Surface and transverse section achene structures ofBolboschoenus maritimus, B. robustus, B. fluviatilis, B. novae-angliae, B. glaucus, and the putative hybridsB. maritimus × B. rubustus andB. glaucus × B. maritimus from North America are described and illustrated with SEM micrographs. The same three type of pericarp structure reported by Browning and Gordon-Gray (1993) from southern Africa were found except that the “type 2” (B. glaucus) was not matched precisely. Intermediate achene structures and other evidence support both the hybrid origin ofB. novae-angliae (=B. fluviatilis × B. robustus) and hybridization betweenB. glaucus andB. maritimus. The new combinationBolboschoenus novae-angliae is made.     

Phylogenetic, morphological, and chemotaxonomic incongruence in the North American endemic genus Echinacea

The study of recently formed species is important because it can help us to better understand organismal divergence and the speciation process. However, these species often present difficult challenges in the field of molecular phylogenetics because the processes that drive molecular divergence can lag behind phenotypic divergence. In the current study we show that species of the recently diverged North American endemic genus of purple coneflower, Echinacea, have low levels of molecular divergence. Data from three nuclear loci and two plastid loci provide neither resolved topologies nor congruent hypotheses about species-level relationships. This lack of phylogenetic resolution is likely due to the combined effects of incomplete lineage sorting, hybridization, and backcrossing following secondary contact. The poor resolution provided by molecular markers contrasts previous studies that found well-resolved and taxonomically supported relationships from metabolic and morphological data. These results suggest that phenotypic canalization, resulting in identifiable morphological species, has occurred rapidly within Echinacea. Conversely, molecular signals have been distorted by gene flow and incomplete lineage sorting. Here we explore the impact of natural history on the genetic organization and phylogenetic relationships of Echinacea.     

Within-population genetic structure and clonal diversity of a threatened endemic metallophyte, Viola calaminaria (Violaceae)

We studied the within-population genetic structure and the clonality extent of Viola calaminaria, a rare endemic species of calamine soils, by means of RAPD markers in two populations (one recent and one ancient) with expected harsh and heterogeneous heavy-metal stress. At a very local scale (0.2-3 m), clonal propagation was detected in both populations, but the levels of clonal diversity were high (number of genets/number of ramets sampled = 0.9 [recent] and 0.76 [ancient]) and the maximal observed extension of the clones was 0.4 m. This indicated that clonality is not, for the species, an important mode of propagation and that clonal growth cannot be interpreted as a strategy for propagating or perpetuating adapted genotypes under harsh ecological constraints. Spatial autocorrelations revealed a significant (P < 0.001) negative value of correlogram slope in the two populations even when a single individual per clone was considered (i.e., analysis at the genet level). We conclude that spatial genetic structure at a very local scale reflects limited gene flow due to restricted seed dispersal rather than variation in clonal pattern in response to environmental heterogeneity. At a larger scale (2-30 m), spatial autocorrelations revealed a positive (P < 0.001) correlation at < 3 m and a random pattern at larger distances for the two populations. This suggested a patchy distribution of the genetically linked individuals associated with a disrupted pattern at a longer distance probably due to gene flow by pollen dispersal and a seed bank effect. The implications for the conservation of V. calaminaria are discussed.     

A satellite DNA evolutionary analysis in the North American endemic dioecious plant Rumex hastatulus (Polygonaceae)

We studied the evolution of RAE180 satellite DNA family in the North American endemic dioecious plant Rumex hastatulus. In this species, the Texas race is characterized by a single XX/XY sex chromosome system, whereas the North Carolina race has evolved a derived complex XX/XY(1)Y(2) sex chromosome system. RAE180 repeats were autosomic and poorly represented (2 × 10(-4)% of the genome) with no differences between individuals of different genders or different races of R. hastatulus. In fact, the sex chromosomes of the North Carolina race are still euchromatic, and they have not accumulated satellite DNA sequences, which contrasts with that occurring in the rest of dioecious XX/XY(1)Y(2) Rumex species. In R. hastatulus, we detected the existence of three RAE180 subfamilies. Notwithstanding, while in the Texas race the TX1/NC1 subfamily is the most frequent, the TX2/NC2 subfamily is the most abundant in the North Carolina race. Additionally, the third, less represented subfamily (TX3/NC3) appears currently as relict sequences in both genomes. A common feature of RAE180 satellite is the sudden replacement of one sequence variant by another in different species (or populations as in R. hastatulus races). Thus, the phylogenetic analysis of RAE180 repeats from six dioecious Rumex species supports the "library" hypothesis. According to this hypothesis, we assume that a set of divergent RAE180 variants were present in the ancestral genome of dioecious Rumex species, from which novel tandem arrays originated by the amplification of different variants in different lineages. Differential levels of RAE180 satellite DNA amplification in each lineage, at different evolutionary times, and in different chromosomal positions gave rise to differential patterns of sequence evolution.     

Population genetic structure of the South American species Hypochaeris lutea (Asteraceae)

The genus Hypochaeris has a recent evolutionary history caused by long‐distance dispersal in conjunction with adaptive radiation in the South American continent. Hypochaeris lutea is a perennial herb that grows mostly at altitudes of around 1000 m in cold swamps of the southern regions of Brazil. We investigated the amplified fragment length polymorphism (AFLP) in 270 individuals representing 11 Brazilian populations of H. lutea to elucidate the population genetic structure of this species. The frequencies of polymorphic loci and gene diversity ranged from 83.42% to 91.66% and from 0.26 to 0.34, respectively. Analysis of molecular variance revealed that most of the genetic variability was found within (76.67%) rather than among (23.3%) populations, agreeing with the pattern of genetic distribution within and among populations observed in other allogamous species of Hypochaeris. A Mantel test showed no correlation between genetic and geographic distances when all populations were considered. Simulations performed using a Bayesian approach consistently identified two clusters with different admixture proportions of individuals, as also revealed by a UPGMA dendrogram of populations. The pattern of genetic structure observed in H. lutea is consistent with a process of successive colonization events by long‐distance dispersal resembling the rapid and recent radiation that has been proposed to explain the origin of the South American species of Hypochaeris.     

Spatial genetic structure in populations of the terrestrial orchid Orchis cyclochila (Orchidaceae)

Orchid seeds are minute, dust-like, wind-borne and, thus, would seem to have the potential for long-distance dispersal. Based on this perception, one may predict near-random spatial genetic structure within orchid populations. In reality we do not know much about seed dispersal in orchids and the few empirical studies of fine-scale genetic structure have revealed significant genetic structure at short distances (< 5m), suggesting that most seeds of orchids fall close to the maternal plant. To obtain more empirical data on dispersal, Ripley’s L(d)-statistics, spatial autocorrelation analyses (coancestry, f_ij analyses) and Wright’s F statistics were used to examine the distribution of individuals and the genetic structure within two populations of the terrestrial orchid Orchis cyclochila in southern Korea. High levels of genetic diversity (H_e = 0.210) and low between-population variation were found (F_ST = 0.030). Ripley’s L(d)-statistics indicated significant aggregation of individuals, and patterns varied depending on populations. Spatial autocorrelation analysis revealed significant positive genetic correlations among individuals located <1 m, with mean f_ij values expected for half sibs. This genetic structure suggests that many seeds fall in the immediate vicinity of the maternal plant. The finding of significant fine-scale genetic structure, however, does not have to preclude the potential for the long distance dispersal of seeds. Both the existence of fine-scale genetic structure and low F_ST are consistent with a leptokurtic distribution of seed dispersal distances with a very flat tail.     

Changes in genetic structure of North American Bythotrephes populations following invasion from Lake Ladoga, Russia

1. We used allozyme electrophoresis to compare Bythotrephes longimanus (Crustacea: Onychopoda: Cercopagididae) from recently founded North American populations with those from native European populations, and to examine changes in genetic structure of North American populations over time.
2. The genetic structure of North American populations in 1996 was similar to that of European populations, because of the disappearance of founder effects which distinguished North American from European populations in 1989.
3. The Lake Ladoga, Russia population was more closely related to North American populations than to other European populations, consistent with non-genetic evidence implicating Lake Ladoga as the source of North American populations.
4. Our results provide additional evidence of the presence of an invasion corridor that allows Urasian and Ponto-Caspian species to be introduced into North American freshwater ecosystems, and show that founder effects can erode over time following establishment of invasive species.     

Population genetic structure of a California endemic branchiopod,Branchinecta sandiegonensis

Branchinecta sandiegonensis (Crustacea: Anostraca) is a narrow range endemic fairy shrimp discontinuously distributed in ephemeral pools on coastal mesas in San Diego County, USA. Ten populations across the range of the species were subjected to allozyme analysis for eleven loci. The species exhibits low variability (P₉₅ =9.1–45.5) and one third of the loci tested did not conform to Hardy-Weinberg equilibrium expectations. The species also exhibited a high degree of genetic differentiation between populations. F _ST values (fixation index) for most pairs of populations were above 0.25 (0.036–0.889).Low genetic variability and high genetic structure may result from low gene flow and founder effects due to habitat fragmentation and the lack of potential vectors for cyst dispersal. The unpredictable rainfall of the region also creates potential for variable population sizes which could affect structure and variability. This revised version was published online in August 2006 with corrections to the Cover Date.     

Phlebotomine fauna (Diptera: Psychodidae) and species abundance in an endemic area of American cutaneous leishmaniasis in southeastern Minas Gerais, Brazil

This study was undertaken to identify the phlebotomine fauna and species abundance in domiciliary and peridomiciliary (hen-house and guava-tree) environments, on a lake shore, a cultivated area of coffee and banana, and a forested area of Concei??o da Aparecida municipality, southeastern the state of Minas Gerais, to provide information for the control and epidemiological surveillance of leishmaniasis in this area. The captures were carried out monthly between May 2001 and November 2002, with automatic light and Shannon traps. A total of 1444 sand flies were captured, 951 (76.5%) with automatic light traps and 493 (23.5%) with the Shannon trap. Thirteen species were captured, the most frequent being Nyssomyia whitmani (62.7%), Migonemyia migonei (21.4%), Pintomyia fischeri (6.9%), and Evandromyia lenti (3.6%). Species abundance was determined using the automatic light traps installed in the six environments. The most abundant species according to the standardized index of species abundance were Ny. whitmani (1.0) and Mg. migonei (0.82). In view of the dominance of these two species, known vectors of cutaneous leishmaniasis in other Brazilian areas, their participation in the transmission of the disease in this county is suggested. The diversity and evenness indexes in the domicile were the lowest due to the high frequency (83%) of Ny. whitmani. The capture of Lutzomyia longipalpis, rarely recorded in the south-eastern and southern regions of Minas Gerais, is also noteworthy.