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Nevirapine (NVP) is an effective drug for the treatment of HIV infections, but its use is limited by a high incidence of severe skin rash and liver injury. 12-Hydroxynevirapine (12-OH-NVP) is the major metabolite of nevirapine. There is strong evidence that the sulfate of 12-OH-NVP is responsible for the skin rash. While several cytosolic sulfotransferases (SULTs) have been shown to be capable of sulfating 12-OH-NVP, the exact mechanism of sulfation in vivo is unclear. The current study aimed to clarify human SULT(s) and human organs that are capable of sulfating 12-OH-NVP and investigate the metabolic sulfation of 12-OH-NVP using cultured HepG2 human hepatoma cells. Enzymatic assays revealed that of the thirteen human SULTs, SULT1A1 and SULT2A1 displayed strong 12-OH-NVP-sulfating activity. 1-Phenyl-1-hexanol (PHHX), which applied topically prevents the skin rash in rats, inhibited 12-OH-NVP sulfation by SULT1A1 and SULT2A1, implying the involvement of these two enzymes in the sulfation of 12-OH-NVP in vivo. Among five human organ cytosols analyzed, liver cytosol displayed the strongest 12-OH-NVP-sulfating activity, while a low but significant activity was detected with skin cytosol. Cultured HepG2 cells were shown to be capable of sulfating 12-OH-NVP. The effects of genetic polymorphisms of SULT1A1 and SULT2A1 genes on the sulfation of 12-OH-NVP by SULT1A1 and SULT2A1 allozymes were investigated. Two SULT1A1 allozymes, Arg37Asp and Met223Val, showed no detectable 12-OH-NVP-sulfating activity, while a SULT2A1 allozyme, Met57Thr, displayed significantly higher 12-OH-NVP-sulfating activity compared with the wild-type enzyme. Collectively, these results contribute to a better understanding of the involvement of sulfation in NVP-induced skin rash and provide clues to the possible role of SULT genetic polymorphisms in the risk of this adverse reaction., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier Inc.)

This review summarizes the development of population genetics and population genomics studies of forest trees in Slovakia during the past 40 years. Various protein and DNA markers have been applied during this period to address several topics in evolutionary genetics and biogeography of trees: allozymes, uniparentally inherited chloroplast and mitochondrial markers, simple sequence repeats and single nucleotide polymorphisms. The main object of studies of phylogeny and postglacial migration were Fagus sylvatica s.l . and eastern-Mediterranean firs ( Abies Mill. section Abies ), where the divergence of genetic lineages (species and subspecific taxa) in time, as well as colonization of the current ranges during the Holocene were reconstructed. The studies on intraspecific gene flow and homoploid hybridization focused on hybrid swarms Pinus sylvestris/P. mugo and firs. Unusual maternal inheritance of chloroplast DNA was revealed in P. mugo × P. sylvestris crosses. Contrasting geographical structures of hybrid zones were revealed in wind-dispersed vs. animal-dispersed trees. Within the studies of adaptation, signals of selection were identified both in field observations and common-garden experiments on Picea abies, F. sylvatica and A. alba . Perspectives of ongoing research employing next-generation sequencing were shortly outlined., Competing Interests: Conflicts of interest/competing interestsThe authors do not perceive any conflict of interests., (© Slovak Academy of Sciences 2021.)

Amphibians are the most vulnerable vertebrates to biodiversity loss mediated by habitat destruction, climate change and diseases. Informed conservation management requires improving the taxonomy of anurans to assess reliably the species' geographic range. The genus Odontophrynus that is geographically refined to Argentina, Bolivia, Brazil, Uruguay and Paraguay includes currently 12 nominal species with many populations of uncertain taxonomic assignment and subsequently unclear geographic ranges. In this study, we applied integrative taxonomic methods combining molecular (mitochondrial 16S gene), allozyme, morphological and bioacoustic data to delimit species of the genus Odontophrynus sampled from throughout Argentina where most species occur. The combined evidence demonstrates one case of cryptic diversity and another of overestimation of species richness. The populations referred to as O. americanus comprise at least three species. In contrast, O. achalensis and O. barrioi represent junior synonyms of the phenotypically plastic species O. occidentalis . We conclude that each of the four species occurring in Argentina inhabits medium to large areas. The Red List classification is currently "Least Concern". We also propose a phylogenetic hypothesis for the genus and associated genera Macrogenioglottus and Proceratophrys (Odontophrynidae)., Competing Interests: The authors declare that they have no competing interests.

The effect of radiation pollution on genetic variation in natural populations of Melandrium album was investigated at the head part of the East-Ural Radioactive Trace (EURT) and background areas. The highest genetic differentiation estimated using F ST was revealed between compared pairs of the background and impact samples in populations of M. album. The highest rate of polymorphism was observed at the closest to nuclear accident, Impact-1 site. The unique alleles (Mdh-3 104 , Pgi-2 106 , Lap 105 , Mdh-2 96 , and Dia 94 ) were discovered at the EURT. Individuals from chronically low-level irradiated sites were genetically closer than to plants from background sites using Nadhdh locus. The increase of the frequency of unique homozygous and heterozygous genotypes was identified in populations of M. album growing under chronic radiation exposure conditions. The largest contribution to the group of unique heterozygous genotypes at the EURT was made by three loci - Lap, Pgi-2, and Nadhdh; the main role in interpopulation differentiation of samples was made by the alleles Sod-2 115 , Skdh 100 , and Nadhdh 100 . Our results provide evidence for the correlation between the increase of genetic variation other than the «genetic erosion» and chronic radiation exposure factor in natural plant populations.

Premise of the Study: Cryptic species are a challenge for botanists and taxonomists. To improve species delineation in the genus Botrychium (Ophioglossaceae), which includes multiple instances of allotetraploid speciation, we examined a cryptic species complex using genetics and morphology., Methods: We sampled species in the B. matricariifolium complex, concentrating on the Upper Peninsula of Michigan and including multiple proposed morphospecies. We analyzed over 1500 samples using 10 enzyme systems, measured 42 quantitative and qualitative morphological characters for over 650 individuals, and analyzed 145 samples using AFLPs. We tested for diagnostic enzymes in the morphospecies and calculated the correlation between morphological and genetic distances to determine whether putatively distinct morphotypes warrant taxonomic recognition., Key Results: Allozyme allelic variation corresponded loosely to some morphotypes of B. matricariifolium, but with lower genetic distinction among them than found between B. matricariifolium and B. michiganense. Botrychium michiganense contains unique alleles, indicating a different hybrid origin from that of B. matricariifolium and supporting its status as a genetically distinct species., Conclusions: We showed that B. acuminatum morphology and genetics are accommodated taxonomically within B. matricariifolium; B. matricariifolium and B. michiganense likely represent hybridization events between related species; and morphotypes within B. matricariifolium likely represent repeated hybridization events between the same two parental species. These hybridizations have resulted in the array of morphotypes observed by field botanists. By helping to identify diagnostic morphological characters, genetic analyses also help us understand and resolve morphological variation observed in the field., (© 2016 Botanical Society of America.)

The Ross Sea, Eastern Antarctica, is considered a "pristine ecosystem" and a biodiversity "hotspot" scarcely impacted by humans. The sibling species Contracaecum osculatum sp. D and C. osculatum sp. E are anisakid parasites embedded in the natural Antarctic marine ecosystem. Aims of this study were to: identify the larvae of C. osculatum (s.l.) recovered in fish hosts during the XXVII Italian Expedition to Antarctica (2011-2012); perform a comparative analysis of the contemporary parasitic load and genetic variability estimates of C. osculatum sp. D and C. osculatum sp. E with respect to samples collected during the expedition of 1993-1994; to provide ecological data on these parasites. 200 fish specimens (Chionodraco hamatus, Trematomus bernacchii, Trematomus hansoni, Trematomus newnesi) were analysed for Contracaecum sp. larvae, identified at species level by allozyme diagnostic markers and sequences analysis of the mtDNA cox2 gene. Statistically significant differences were found between the occurrence of C. osculatum sp. D and C. osculatum sp. E in different fish species. C. osculatum sp. E was more prevalent in T. bernacchii; while, a higher percentage of C. osculatum sp. D occurred in Ch. hamatus and T. hansoni. The two species also showed differences in the host infection site: C. osculatum sp. D showed higher percentage of infection in the fish liver. High genetic variability values at both nuclear and mitochondrial level were found in the two species in both sampling periods. The parasitic infection levels by C. osculatum sp. D and sp. E and their estimates of genetic variability showed no statistically significant variation over a temporal scale (2012 versus 1994). This suggests that the low habitat disturbance of the Antarctic region permits the maintenance of stable ecosystem trophic webs, which contributes to the maintenance of a large populations of anisakid nematodes with high genetic variability.

We found low genetic differentiation between two temporal samples of Fasciola hepatica (2006 and 2008) collected from nine sheep of the same flock that shared the same pasture for at least 2 years. However, each sample, represented by four and five infrapopulations respectively, showed strong heterozygote deficits regarding Hardy-Weinberg expectations and a high degree of genetic structure at infrapopulation level. This is an unexpected result since genetic drift should increase temporal variation among years. Our findings are most likely explained by the fact that the parasite can survive many years in the definitive host. Temporal gene flow favored by high longevity probably increases levels of genetic variability of the population but could also contribute to the observed heterozygote deficits within temporal samples and infrapopulations if it favors the Wahlund effect. Despite the homogenizing effect of gene flow, the high genetic divergence observed between infrapopulations is most likely a consequence of strong genetic drift associated to the complexity of the life cycle., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Widespread species with morphologically and ecologically differentiated populations are key to understand speciation because they allow investigating the different stages of the continuous process of population divergence. The alpine newt, Ichthyosaura alpestris, with a range that covers a large part of Central Europe as well as isolated regions in all three European Mediterranean peninsulas, and with strong ecological and life-history differences among populations, is an excellent system for such studies. We sampled individuals across most of the range of the species, and analyzed mitochondrial (1442 bp) and nuclear (two nuclear genes -1554 bp- and 35 allozyme loci) markers to produce a time-calibrated phylogeny and reconstruct the historical biogeography of the species. Phylogenetic analyses of mtDNA data produced a fully resolved topology, with an endemic, Balkan clade (Vlasina) which is sister to a clade comprising an eastern and a western group. Within the former, one clade (subspecies I. a. veluchiensis) is sister to a clade containing subspecies I. a. montenegrina and I. a. serdara as well as samples from southern Romania, Bosnia-Herzegovina, Serbia and Bulgaria (subspecies I. a. reiseri and part of I. a. alpestris). Within the western group, populations from the Italian peninsula (subspecies I. a. apuana and I. a. inexpectata) are sister to a clade containing samples from the Iberian Peninsula (subspecies I. a. cyreni) and the remainder of the samples from subspecies I. a. alpestris (populations from Hungary, Austria, Poland, France, Germany and the larger part of Romania). Results of (∗)BEAST analyses on a combined mtDNA and nDNA dataset consistently recovered with high statistical support four lineages with unresolved inter-relationships: (1) subspecies I. a. veluchiensis; (2) subspecies I. a. apuana+I. a. inexpectata; (3) subspecies I. a. cyreni+part of subspecies I. a. alpestris (the westernmost populations, plus most Romanian populations); and (4) the remaining populations, including subspecies I. a. serdara, I. a. reiseri and I. a. montenegrina and part of subspecies I. a. alpestris, plus samples from Vlasina. Our time estimates are consistent with ages based on the fossil record and suggest a widespread distribution for the I. alpestris ancestor, with the split of the major eastern and western lineages during the Miocene, in the Tortonian. Our study provides a solid, comprehensive background on the evolutionary history of the species based on the most complete combined (mtDNA+nDNA+allozymes) dataset to date. The combination of the historical perspective provided by coalescent-based analyses of mitochondrial and nuclear DNA variation with individual-based multilocus assignment methods based on multiple nuclear markers (allozymes) also allowed identification of instances of discordance across markers that highlight the complexity and dynamism of past and ongoing evolutionary processes in the species., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Short-finned squid Illex argentinus (Omastrephidae) is an economically important species which supports a long-term and intensive commercial fishery in the South-West Atlantic Ocean. In this study, the genetic variability of allozymes (40 loci) and mitochondrial regions, COI (556 bp) and 16S rDNA (439 bp), were addressed in the two most important species' spawning stocks, the Summer Spawning Stock (SSS) and the South Patagonic Stock (SPS). Five out of 20 polymorphic allozyme loci were polymorphic at 95% criterion and heterozygosity levels were low. The concatenated analysis of mitochondrial molecular markers revealed high to low values of haplotype and nucleotide diversity, respectively. Nuclear and mitochondrial data revealed no significant genetic differences between these two spawning stocks. Our results and the biological characteristics of I. argentinus deal with a no simple genetic structure of populations described as spatial and temporal chaotic patchiness. SSS and SPS spawning stocks of I. argentinus could not be elevated to the status of genetic stocks., (Copyright © 2014 Académie des sciences. Published by Elsevier SAS. All rights reserved.)

• Premise of the study: Geographic isolation due to discontinuities of suitable habitat may have significant effects on the genetic structure of plant populations. Even within a few kilometers, physical barriers to gene flow may lead to considerable genetic differentiation among populations.• Methods: Sedum ussuriense is a boreal species that in Korea occurs only in four valleys separated by mountain ranges in Juwangsan National Park and its vicinity (a range of ∼15 km). Its congener S. kamtschaticum, by contrast, co-occurs in the four valleys but also on the intervening mountains. Using 12 allozyme loci, we comparatively assessed genetic variability and structure in 12 population pairs of the two stonecrops.• Key results: While we found high and comparable levels of within-population genetic variation for the two species, among-population divergence was significantly higher in S. ussuriense (F ST = 0.261 vs. F ST = 0.165). Sedum ussuriense also showed a much higher percentage of among-valley variation (19%) than S. kamtschaticum (4%).• Conclusions: High levels of genetic diversity in the two Sedum species are consistent with the previous hypothesis that mountains of the Korean Peninsula served as glacial refugia for many boreal species. Given that the two congeners have similar life-history traits, the lower among-population differentiation in S. kamtschaticum is attributable to its higher abundance and more continuous distribution in the study area. This study confirms the central role of geographic isolation in the genetic structure of plant species even at very small scales., (© 2014 Botanical Society of America, Inc.)

Boechera perstellata is an endangered plant found only in middle Tennessee and north central Kentucky. After sampling 4 Tennessee and 3 Kentucky populations, genetic variability and population structure were examined for this species using isozymes, chloroplast DNA, and microsatellites (averaging 35, 29, and 27 individuals per population per class of marker, respectively). The only genetic variability detected for 23 isozymes was a fixed difference between Tennessee and Kentucky populations at 1 locus. Fixed differences between populations of the 2 states were also observed for 3 chloroplast markers. Polymorphism at 19 nuclear microsatellites was 74% at the species level and averaged 21% at the population level. However, observed heterozygosity was extremely low in all populations, ranging from 0.000 to 0.005. High F IS values (0.93) suggest that Boechera perstellata is a primarily selfing species. Tennessee populations have more genetic diversity than Kentucky populations of B. perstellata. Microsatellite markers revealed substantial genetic divergence between the states and genetic differences among populations within each state. Analysis of molecular variance indicates that most variability in this species occurs between the 2 states (49%) and among populations within each state (42%), with relatively little variation found within populations (9%). These data indicate that there is very little gene flow among populations of B. perstellata and that it is important to protect as many populations as possible in order to conserve the genetic diversity of this rare species.

Background and Aims: Despite the great importance of autopolyploidy in the evolution of angiosperms, relatively little attention has been devoted to autopolyploids in natural polyploid systems. Several hypotheses have been proposed to explain why autopolyploids are so common and successful, for example increased genetic diversity and heterozygosity and the transition towards selfing. However, case studies on patterns of genetic diversity and on mating systems in autopolyploids are scarce. In this study allozymes were employed to investigate the origin, population genetic diversity and mating system in the contact zone between diploid and assumed autotetraploid cytotypes of Vicia cracca in Central Europe., Methods: Four enzyme systems resolved in six putative loci were investigated in ten diploid, ten tetraploid and five mixed-ploidy populations. Genetic diversity and heterozygosity, partitioning of genetic diversity among populations and cytotypes, spatial genetic structure and fixed heterozygosity were analysed. These studies were supplemented by a pollination experiment and meiotic chromosome observation., Key Results and Conclusions: Weak evidence of fixed heterozygosity, a low proportion of unique alleles and genetic variation between cytotypes similar to the variation among populations within cytotypes supported the autopolyploid origin of tetraploids, although no multivalent formation was observed. Tetraploids possessed more alleles than diploids and showed higher observed zygotic heterozygosity than diploids, but the observed gametic heterozygosity was similar to the value observed in diploids and smaller than expected under panmixis. Values of the inbreeding coefficient and differentiation among populations (ρST) suggested that the breeding system in both cytotypes of V. cracca is mixed mating with prevailing outcrossing. The reduction in seed production of tetraploids after selfing was less than that in diploids. An absence of correlation between genetic and geographic distances and high differentiation among neighbouring tetraploid populations supports the secondary contact hypothesis with tetraploids of several independent origins in Central Europe. Nevertheless, the possibility of a recent in situ origin of tetraploids through a triploid bridge in some regions is also discussed.

Species delineation remains one of the most challenging tasks in the study of biodiversity, mostly owing to the application of different species concepts, which results in contrasting taxonomic arrangements. This has important practical consequences, since species are basic units in fields like ecology and conservation biology. We here review molecular genetic evidence relevant to the systematics of toads in the Bufo bufo species group (Anura, Bufonidae). Two studies recently published in this journal (Recuero et al., MPE 62: 71-86 and García-Porta et al., MPE 63: 113-130) addressed this issue but reached opposing conclusions on the taxonomy of the group (four versus two species). In particular, allozyme data in the latter paper were interpreted as evidence for hybridization across species (between B. bufo-B. spinosus and B. bufo-B. verrucosissimus). We tested claims for hybridization through re-analysis of allozyme data for individuals instead of populations, to be able to distinguish between sympatry with and without admixture, and found no evidence of hybridization across taxa. We propose alternative explanations for the observed patterns that García-Porta et al. (2012) failed to consider. In the absence of unequivocal evidence for hybridization and introgression, we reject the proposal to downgrade Bufo spinosus and Bufo verrucosissimus to the subspecies level., (Copyright © 2013 The Authors. Published by Elsevier Inc. All rights reserved.)

Premise of the Study: Drosera peltata var. nipponica, an element of the East Asia warm-temperate vegetation, and D. rotundifolia, a widely distributed boreal species, reach one of their northernmost and southernmost limits, respectively, on the Korean Peninsula. Because the Last Glacial Maximum (LGM)-Holocene dynamics of warm-temperate and boreal paleovegetation differed considerably on the Peninsula, the population history of these two sundews is expected to be different, leaving differential imprints in their genetic structure., Methods: We investigated population genetic structure of D. peltata var. nipponica and D. rotundifolia in South Korea (10 populations of each for 20 allozyme loci) to infer their population history in this region. In addition, we compared the genetic variation harbored in the two sundews to those reported for other carnivorous and wetland plants., Key Results: Drosera peltata var. nipponica showed no genetic diversity, whereas D. rotundifolia exhibited extremely low within-population variation (He = 0.005) and considerable among-population divergence (FST = 0.817)., Conclusions: Our results suggest that extant populations of D. peltata var. nipponica likely originated from a single ancestral population from southern Japan or southern China through postglacial dispersal. On the contrary, D. rotundifolia probably survived the LGM in situ, with extant populations derived from either one or several small source populations. We argue that separate conservation strategies should be employed, given that the two taxa have different ecological and demographic traits and harbor different levels of genetic diversity.

Spores of homosporous ferns are small, wind-borne and thus have the potential for long-distance dispersal. This common perception has led to a prediction of near-random spatial genetic structure within fern populations. Spore dispersal and spore bank studies, however, indicate that most spores fall close to the maternal plant (< 5 m), supporting a prediction of significant fine-scale genetic structure (FSGS) within populations. To determine which of these two hypotheses is more likely to occur in nature, we measured inbreeding and quantified the spatial distribution of individuals and allozyme-based genotypes using spatial autocorrelation methods within four populations of the fern Cyrtomium falcatum in southern South Korea. Inbreeding levels were low, and all populations exhibited significant aggregation of individuals and strong FSGS. The present results support the second hypothesis, and the substantial FSGS in C. falcatum could reflect the unique features of most homosporous ferns (outcrossing mating systems that lead a majority of spores to occur at short distances and a very limited dispersal distance of male gametes). Although fern spores are physically analogous to orchid seeds, the intensity of FSGS exhibited in C. falcatum is four times stronger than that in 16 terrestrial orchid species., (© 2013 The Authors. New Phytologist © 2013 New Phytologist Trust.)

Various aspects of the genetics of the rat lungworm, Angiostrongylus cantonensis, are reviewed. This nematode has an XX/XO sex-determination mechanism, with the female having a 2n = 12 (XX) and the male 2n = 11 (XO) chromosome constitution. Allozymes (12 loci) exhibit a low proportion of polymorphic loci (P = .08) and low mean heterozygosity (H = 0.43) in specimens from Hawa'i, and no polymorphism or heterozygosity in specimens from Thailand. The phosphoglucomutase-2 (PGM-2) locus exhibits sex-limited expression, with no detectable enzyme activity in the male worms from either location. Based on the 12 allozyme loci, Nei's genetic distance between the Hawa'i and Thailand isolates is D = 0.03. The p-distance (proportion of nucleotide sites) based on cytochrome c oxidase subunit I (COI) is 3.61% between the Thailand and China isolates as well as between Thailand and Hawa'i isolates, and 0.83% between China and Hawa'i isolates. The partial DNA sequences of the 66 kDa protein gene show a great diversity of haplotypes, indicating both inter- and intra-population variation. Intra-specific sequence variation is also found in the internal transcribed spacer regions. For the small-subunit ribosomal RNA (SSU rRNA) gene, two distinct genotypes have been recorded.

Background and Aims Allozyme and reproductive data sets for the Canarian flora are updated in order to assess how the present levels and structuring of genetic variation have been influenced by the abiotic island traits and by phylogenetically determined biotic traits of the corresponding taxa; and in order to suggest conservation guidelines. Methods Kruskal-Wallis tests are conducted to assess the relationships of 27 variables with genetic diversity (estimated by A, P, Ho and He) and structuring (GST) of 123 taxa representing 309 populations and 16 families. Multiple linear regression analyses (MLRAs) are carried out to determine the relative influence of the less correlated significant abiotic and biotic factors on the genetic diversity levels. Key Results and Conclusions The interactions between biotic features of the colonizing taxa and the abiotic island features drive plant diversification in the Canarian flora. However, the lower weight of closeness to the mainland than of (respectively) high basic chromosome number, partial or total self-incompatibility and polyploidy in the MLRAs indicates substantial phylogenetic constraint; the importance of a high chromosome number is feasibly due to the generation of a larger number of linkage groups, which increase gametic and genotypic diversity. Genetic structure is also more influenced by biotic factors (long-range seed dispersal, basic chromosome number and partial or total self-incompatibility) than by distance to the mainland. Conservation-wise, genetic structure estimates (FST/GST) only reflect endangerment under intensive population sampling designs, and neutral genetic variation levels do not directly relate to threat status or to small population sizes. Habitat protection is emphasized, but the results suggest the need for urgent implementation of elementary reproductive studies in all cases, and for ex situ conservation measures for the most endangered taxa, even without prior studies. In non-endangered endemics, multidisciplinary research is needed before suggesting case-specific conservation strategies. The molecular information relevant for conservation should be conserved in a standardized format to facilitate further insight.

Allozyme electrophoresis was used to examine 12 enzymatic systems in two populations of the genus Neoplecostomus from the Paraná River basin. Samples of Neoplecostomus sp. 1 were collected in Paraitinguinha stream of the Tietê River basin, in the municipality of Salesópolis, São Paulo State, and those of Neoplecostomus sp. 2 from São Domingos stream of the Rio Grande River basin, in the municipality of Muzambinho, Minas Gerais State. The genetic variability of the two populations was estimated by Nei's expected heterozygosity and was considered lower than average for populations of freshwater fish. The proportion of polymorphic loci was low (only 5.26% for the locus Idh). The low frequency of heterozygosity for both populations revealed a high fixation of alleles for each locus. Homozygote excess was observed in both populations. The values of Nei's genetic identity and the presence of loci with different allele frequencies in both populations may imply that the two populations belong to different species. The genetic variability between populations was compared to other data for loricariids.

Only very few cases have documented that an increase in connectivity after a period of fragmentation in ecological time has had an effect on the distribution, genetic structure and morphology of stenotopic species. In this study we present an example of clinal variability in a woodland ground beetle as a result of changes in the connectivity of a landscape during the last two centuries. The study area hosts both the nominate form Carabus violaceus s. str. and the subspecies Carabus violaceus purpurascens, which is ranked as a distinct species by some authors. We studied 12 Carabus violaceus populations from a 30 km transect of ancient and recent forests in north-western Germany. We analyzed three polymorphic enzyme loci, classified the elytron sculpture and measured the shape of the aedeagus tip of the specimens. Carabus violaceus showed secondary gradients both in allozyme markers and morphometric characters in our study area. A genetic differentiation of 16% between the populations is high but lies within the range of intraspecific variability in habitat specialists of the genus Carabus. Populations had no significant deficit of heterozygotes. We found many hybrid populations in terms of morphological properties. This study highlights the conservation value of ancient woodland and the consequences of landscape connectivity and defragmentation on the genetic setting of a ground beetle. Moreover, it shows that differences in the external shape of male genitalia do not prevent gene flow within the genus Carabus. Thus, the establishment of species status should not exclusively be based on this property.

Two allopatric morphotypes of the genus Rinelocaria were compared through the allozyme electrophoresis technique: one morphotype, R. pentamaculata, from the Keller River in the middle stretch of the Ivaí River basin and the other, R. aff. pentamaculata, from the São João River in the upper portion of the Ivaí River basin. The morphotype from the São João River was collected upstream from the São João waterfall, which is about 80 m deep. Twelve enzymatic systems (AAT, ADH, EST, GCDH, G3PDH, GPI, IDH, LDH, MDH, ME, PGM and SOD) were analyzed, which allowed to score 22 loci. Only lociAat-2, Est-3 and Mdh-C showed polymorphism. The two samples differed in allele frequencies at the three polymorphic loci. The average expected heterozygosity for all loci was 0.0806 ± 0.0447 in the Keller River sample. For the São João River morphotype, this value was 0.0489 ± 0.0350. Nei' s genetic identity and distance between the two populations were respectively 0.9789 and 0.0213. Wright's F(IS) , F(IT) and F(ST) over all loci were estimated as 0.3121, 0.4021 and 0.1309, respectively. We consider that the two morphotypes represent species in statunascendi.

Two syntopic morphotypes of the genus Hypostomus - H. nigromaculatus and H. cf. nigromaculatus (Atlântico Stream, Paraná State) - were compared through the allozyme electrophoresis technique. Twelve enzymatic systems (AAT, ADH, EST, GCDH, G3PDH, GPI, IDH, LDH, MDH, ME, PGM and SOD) were analyzed, attributing the score of 20 loci, with a total of 30 alleles. Six loci were diagnostic (Aat-2, Gcdh-1, Gpi-A, Idh-1, Ldh-A and Mdh-A), indicating the presence of interjacent reproductive isolation. The occurrence of few polymorphic loci acknowledge two morphotypes, with heterozygosity values He = 0.0291 for H. nigromaculatus and He = 0.0346 for H. cf. nigromaculatus. F(IS) statistics demonstrated fixation of the alleles in the two morphotypes. Genetic identity (I) and distance (D) of Nei (1978) values were I = 0.6515 and D = 0.4285. The data indicate that these two morphotypes from the Atlântico Stream belong to different species.

This study assessed intrinsic differences in tissue quality and growth rate among populations of Metrosideros polymorpha native to sites with a range of soil fertilities. We collected seedlings from three Hawaiian mesic forests that were either phosphorus-limited, nitrogen-limited, or relatively fertile. These individuals were grown in a common garden under a factorial high/low, N/P fertilization regime for 1.5 years and then harvested to determine genetic divergence; aboveground growth rate; and lignin, N, and P concentrations in leaves and roots. Allozyme analyses indicated that the three groups had genetically diverged to some degree (genetic distance = 0.036-0.053 among populations). Relative growth rate did not differ significantly among the populations. Senescent leaves from the fertile-site population had the highest N concentrations (due to low N resorption) and had lower lignin concentrations than plants from the N-limited site. Across treatments, P concentrations in senescent leaves were highest in plants from the fertile and P-limited site. Root tissue quality did not generally differ significantly among populations. Since decomposition rate of senescent leaves in this system is related positively to N concentration and negatively to lignin concentration, senescent leaves from the fertile-site population may have a genetic tendency toward faster decay than the others. The intrinsic qualities of the three populations may provide positive feedbacks on nutrient cycling at each site-nutrient availability may be raised to some degree at the fertile site, and reduced at the N- or P-limited sites. Our results suggest that even a small degree of genetic differentiation among groups can influence traits related to nutrient cycling.

The genetic structure of populations of Schizophyllum commune was inferred from electrophoretic variation among 136 individuals at 11 polymorphic allozyme loci to determine the extent of geographic differentiation in this widespread mushroom species. The majority of the genetic variation was contained within populations; however, considerable genetic differentiation was observed among populations (global G ST = 0.214). Clustering analysis demonstrated that genetic distance was correlated with geographic distance and that a large component of the genetic variation was due to allele frequency differences among populations from the eastern and western hemispheres. Our results also suggest that populations are large and geographically widespread. The lack of fixed genetic differences among intercontinental populations at any of the allozyme loci suggests that long-distance spore dispersal may counter the effects of genetic drift in this cosmopolitan species. These results are contrasted with a previous description of the same collection, in which the mating allele distribution of the species displayed no population substructure at any geographic scale (Raper et al. 1958). Broader implications of this study are that both species and mating allele distributions may not be correlated with long-distance gene flow in basidiomycete fungi., (© 1999 The Society for the Study of Evolution.)

Spatial variation in allelic frequencies at nine allozyme loci were assayed in 20 populations of the crown-of-thorns starfish, Acanthaster planci, collected throughout the Pacific and Indian Oceans. These data were analyzed together with published data, for the same loci, from an additional 19 populations, giving a total sample size of approximately 1800 individuals. There was a marked discontinuity between the Indian and Pacific Ocean populations, but those off Western Australia and from the Southeast Asian region had a strong Pacific affinity. The genetic groups were congruent with the distributions of two color morph groups: gray-green to red-brown forms in the Pacific and a blue to pale red form in the Indian Ocean. These patterns of genetic structure are similar to those described for the starfish Linckia laevigata, which has similar life-history characteristics. Vicariant events may have influenced some populations within the Pacific, but the allozyme data cannot resolve the effects of these events clearly. Patterns of variation within regions were consistent with isolation by distance, but, at larger scales, were obscured by regional vicariance and some outliers, particularly by apparently high levels of gene flow between Japan and the Great Barrier Reef, Australia. Apparent gene flow between population pairs was not closely related to present-day ocean currents. The results demonstrate a strong influence of allopatric separation on genetic divergence at large geographic scales, but also show evidence of slow rates of change in gene frequencies consistent with the large population sizes of this species. Low levels of divergence between groups demonstrate the genetic structure is recent (Pleistocene) and are likely responses to changes in climate and sea level., (© 1999 The Society for the Study of Evolution.)

The sponge species Chondrilla nucula has a simple morphology and a very wide geographical distribution. To verify whether the latter might be an artifact of the former, samples of this species were collected across 10,000 km of its range, in the Mediterranean, the Caribbean, and the southwestern atlantic. The classical (spicule morphology) and molecular (allozymes) systematic approaches were compared, to try to define the geographic limits between populations and detect possible cryptic species. We found five distinct genetic forms within C. nucula that sometimes showed morphological homogeneity and other times plasticity. The difference in size of spicules could not be related to the clear-cut genetic differences, suggesting that the use of spicule sizes for sponge systematics should be reappraised. The population of one of the genetic forms along 3000 km of the Brazilian coast was highly structured (F ST = 0.21; N e m = 0.96). Our results reject the null hypothesis of cosmopolitanism of C. nucula and indicate that the putative worldwide distribution of some marine sponges, and possibly many other benthic invertebrates, may be the result of overly conservative systematics. Cryptic species appear to be particularly prevalent when genera are well defined but species are characterized by only a few morphological characters., (© 1999 The Society for the Study of Evolution.)

The spatial distribution of clonal versus sexual reproduction in plant populations should generally have differing effects on the levels of biparental inbreeding and the apparent selfing rate, produced via mating by proximity through limited pollen dispersal. We used allozyme loci, join-count statistics, and Moran's spatial autocorrelation statistics to separate the spatial genetic structure caused by clonal reproduction from that maintained in sexually reproduced individuals in two populations of Adenophora grandiflora, a perennial herb. Join-count statistics showed that there were statistically significant clustering of clonal genotypes within distances less than 4 m. Both the entire populations and the sets of sexually reproduced individuals exhibited significant spatial autocorrelation at less than about 12 m, and the sexually reproduced individuals are substantially structured in an isolation-by-distance manner, consistent with a neighborhood size of about 50., (© 1999 The Society for the Study of Evolution.)

I examined the effects of historical division and secondary contact between eastern and western varieties of ponderosa pine (Pinus ponderosa Laws Pinaceae) on extant patterns of genetic variation. Fossil and biogeographic evidence both indicate that the current point of contact between these two varieties represents secondary contact following historical separation during the Wisconsin glaciation. Current gene flow was assessed by observing the degree of introgression of paternally inherited cpDNA and maternally inherited mtDNA polymorphisms. Both seeds and pollen are wind dispersed in ponderosa pine. Introgression was primarily from west to east, the direction of the prevailing wind, for both organelles, but introgression of cpDNA far exceeded that of mtDNA. Thus pollen is the main agent of contemporary gene flow between the two varieties. Neither seeds nor pollen showed enough introgression since secondary contact to have homogenized the two gene pools. However, allozyme differentiation was minimal. This calls into question assumptions of selective neutrality for at least some of the markers. Theory predicts that nuclear markers will show a high locus-to-locus variance of F ST following historical separation. This prediction is confirmed by the allozyme data for ponderosa pine, and may provide a useful means of identifying historical separations from allele frequency data., (© 1999 The Society for the Study of Evolution.)

To shed light on the potential effects of xeric/arid versus mesic environments on plant population genetic structure and patterns of gene flow, we have compared allozyme and cpDNA haplotype variation in populations of two closely related, highly outcrossed, and largely wind-dispersed winter annuals of Senecio (Asteraceae). The species form a distinctive zone of parapatric distribution in the Near East by differing in their ecogeographical regimes. Senecio vernalis mainly thrives in the mesic Mediterranean life zone of Israel, whereas S. glaucus inhabits either xeric maritime or arid (semi-) desert sites. Significant differences in allozymic population subdivision among S. vernalis (θ n = 0.04; Nm n = 5.85) and S. glaucus (θ n = 0.12; Nm n = 1.85) largely resulted from topogeographical substructuring present within the latter species. Because of the similarity of within-region estimates of population structure for S. glaucus with those measured among populations of S. vernalis, it appears unlikely that ecological "aridity" factors per se are important in influencing levels of population differentiation in these species. Based on hierarchical F-statistics and tests of isolation by distance, we further conclude that geographical topologies influence the level and mode of nuclear gene flow (via pollen and/or seed) among and within subsets of S. glaucus populations, although without providing a complete barrier to interregional dispersal (dNm reg = 2.16) and without promoting allopatric differentiation via drift. The allozymic data further suggested that S. vernalis and S. glaucus form a zone of secondary contact in the Near East, accompanied by an almost complete interspecific barrier to nuclear gene flow (dnNm sp = 0.253). However, to account for the considerable sharing of cpDNA haplotypes, both at the intra- and interspecific level, it is necessary to invoke either (1) selection acting against alien nuclear but not cytoplasmic DNA; or (2) the sporadic immigration of cpDNA via seed with large homogenizing effects on cytoplasmic population structure over time., (© 1999 The Society for the Study of Evolution.)

Host-plant dependent fitness trade-offs refer to traits that enhance the performance of an insect on one plant species to its detriment on others. Such trade-offs are central to models of sympatric speciation via host shifts, but have proven difficult to empirically demonstrate. Here, we test for host-plant dependent selection on larvae of apple (Malus pumila L.)- and hawthorn (Crataegus mollis L. spp.)-infesting races of Rhagoletis pomonella (Walsh). Samples of larvae were reared in the field and under protective conditions in a garage. Our rationale was that the garage should slow rates of fruit rot relative to the field, relaxing selection pressures associated with declining fruit quality. Four findings emerged from the study. (1) Larvae suffered higher mortality in fruits in the field than the garage. (2) The increase in mortality was greater for larvae in haws. (3) Larvae possessing the alleles Me 100, Acon-2 95, and Mpi 37, three allozymes displaying host-related differentiation in R. pomonella that map to linkage group II in the fly, left fruits earlier than other genotypes. (4) Allele frequencies for Me 100, Acon-2 95, and Mpi 37 were significantly higher in both apple and haw larvae surviving the field versus the garage treatment. Our results suggested that field conditions favored larvae that rapidly developed and left rotting fruits. Since these individuals tended to possess the alleles Me 100, Acon-2 95, and Mpi 37, frequencies of these allozymes were higher in the field. Selection on larvae was directional for Me 100, Acon-2 95, and Mpi 37 (or linked genes) in both host races. We previously showed that these same alleles can be disfavored in the pupal stage, especially in the apple race, where they correlate with premature diapause termination. Fitness trade-offs in Rhagoletis may therefore be due as much to differences in the relative strengths of directional selection pressures acting on different life stages as to disruptive selection affecting any one particular stage. The necessity to consider details of the entire life-cycle highlights one of the many challenges posed to documenting fitness trade-offs for phytophagous insects., (© 1999 The Society for the Study of Evolution.)

The lack of morphological variation in many freshwater invertebrates over vast distances has been cited as evidence for their frequent, long-distance dispersal. This scenario implies that vicariance will be an insignificant determinant of species distributions or diversity. We carried out a phylogeographic and population genetics study of one widespread crustacean group, the North American Daphnia laevis complex. Allozyme and sequence variation of two mtDNA genes (12S and 16S rRNA) clearly indicates the existence of five morphologically cryptic, largely allopatric groups (Daphnia dubia, D. laevis laevis, D. laevis gessneri, D. magniceps magniceps, and D. magniceps pacifica ssp. n.). Within each of these groups, there is weak or no genetic differentiation over large geographic areas suggesting their recent long-distance dispersal. The present-day distributions and phylogeography of the regional groups suggests the occurrence of both deep and shallow vicariance events. Although divergence times from mtDNA sequences do indicate both deep and shallow divergences, these estimates are incongruent with their proposed vicariance times. The results show that even within closely related freshwater invertebrates, a complex biogeography exists, whose analysis is made difficult by long-distance dispersal, cryptic endemism, and pseudocongruence., (© 1998 The Society for the Study of Evolution.)

Life history should relate to gene flow (Nm) through its influence on dispersal and effective population size. Comparative studies designed to elucidate this relationship must contend with historical events that can yield misleading estimates of gene flow and statistical problems associated with inclusion of life-history traits correlated with phylogeny. We studied the relationships of life-history characters and gene flow in 15 species of darters, a monophyletic group of stream fishes. Populations of coexisting species were sampled in three geographic regions with different Pleistocene glaciation histories. Gene flow was estimated indirectly from allozymes using two methods, 8 and private alleles. Isolation-by-distance was also tested using regression of pairwise estimates of gene flow (M̂) on distance. Theta and private-alleles methods produced congruent estimates of Nm, except in a study region hypothesized to have been historically fragmented and then united following Pleistocene glaciation. A relatively weak association between life-history traits and Nm (based on θ) was observed when species from the historically fragmented region were included in stepwise regression analysis, because Nm was low despite life-history differences among taxa in this region. Excluding observations from this region produced stronger associations between clutch size and Nm (r 2 = 0.57), and between female size, egg size, and Nm (r 2 = 0.95). Additional analyses that corrected for female body size and phylogenetic nonindependence agreed that darters with high fecundity and small eggs exhibited high gene flow, whereas darters with small clutches and large eggs had low gene flow. The latter combination of life-history traits primarily is exhibited in species from headwater habitats where parental investment presumably confers survivorship on offspring. Reduced gene flow and genetic divergence among demes appear to be evolutionary consequences of this strategy., (© 1998 The Society for the Study of Evolution.)

Two recently diverged northeastern Pacific sibling snail species, Nucella ostrina and N. emarginata, currently inhabit adjacent zoogeographic provinces. Their distributions overlap in central California to the north of a major faunal boundary at Point Conception, California (PC). To test the hypothesis that modern sympatry is due to a recent northward range expansion by N. emarginata, I analyzed the population structures of both species with nuclear (allozyme) and mitochondrial DNA (mtDNA) markers. Populations of N. emarginata in the region of overlap exhibit significantly lower heterozygosity and allelic diversity than either populations to the south of PC or populations of N. ostrina. A single mtDNA haplotype characterizes all but one population of N. emarginata sampled in this region, but no haplotype to the south of PC is found at more than one locality. MtDNA haplotypes and allozyme allele frequencies also indicate monophyly of central California populations of N. emarginata. Sharp differences in allelic diversity over small geographic distances may reflect the action of natural selection, but because both nuclear and mtDNA markers display concordant patterns, a range expansion across PC best explains patterns of genetic variation in N. emarginata. Allozymes and mtDNA also reveal that the geologically older N. ostrina is paraphyletic with respect to N. emarginata. This pattern is consistent with, but not indicative of, a peripheral isolation model of speciation. Low genetic diversity is also expected if a significant bottleneck occurred at speciation. However, low allelic diversity is not universal throughout the geographic range of N. emarginata; high allelic diversity at the southern end of the distribution of N. emarginata suggests that in the past N. emarginata has been geographically restricted much further south than PC. A northward range expansion across PC by N. emarginata may thus represent only the most recent postglacial movement by the species. The thermal and oceanographic discontinuities found at PC may not have been directly involved in geographic isolation if N. emarginata originated much further south of this modern boundary. Despite uncertainty regarding the exact spatial distribution of populations at speciation, genetic data indicate that even though N. ostrina and N. emarginata currently exhibit a broad range of geographic overlap, speciation was likely allopatric and was initiated by physical isolation of populations in different zoogeographic provinces., (© 1998 The Society for the Study of Evolution.)

Recent studies of intraspecific phylogeography have suggested that the geographic location of genetic discontinuities, or phylogeographic breaks, may frequently coincide with biogeographic boundaries. The concordance is hypothesized to reflect similarity in the processes governing species boundaries and intraspecific lineage boundaries. This concordance has not, however, been widely tested. In the case of the Point Conception biogeographic boundary between the Oregonian and Californian marine biotas, only the supralittoral copepod Tigriopus californicus has been found to have a coincident phylogeographic break. Here I show that the apparent relationship between this break and Point Conception was, in fact, an artifact of insufficient geographic sampling. Mitochondrial DNA analyses of T. californicus populations between Morro Bay and San Diego reveal at least five equally deep phylogeographic breaks in the region (where only one biogeographic boundary is recognized). Limited nuclear DNA sequence data and allozyme data also support the occurrence of multiple genetic discontinuities along this geographic range. Lack of one-to-one correspondence between intraspecific phylogeography and biogeographic boundaries indicates that the processes affecting the genetic differentiation of populations of T. californicus differ from those responsible for determining species distributional limits at the Point Conception biogeographic boundary. A review of genetic data from other species also fails to provide evidence for concordance of biogeography and intraspecific phylogeography across Point Conception. I suggest that the concordance of phylogeography with biogeography will only be pronounced where the biogeographic boundary separates biotas that are phylogenetically related. The numerous cases of interspecific hybrid zones in the region of Cape Canaveral, for example, indicate that many sister-species pairs occur across this biogeographic boundary. Such hybrid zones are not common at Point Conception, and there appears to be no cases of intraspecific phylogeographic breaks associated with this well-recognized biogeographic boundary., (© 1998 The Society for the Study of Evolution.)

Comparisons of cytoplasmic and nuclear diversity within and among natural plant populations have the potential to distinguish the relative influences of seed and pollen dispersal on contemporary gene flow, or alternatively, may permit inferences of the colonization history of a species via seed. We examined patterns of cpDNA and allozyme variation in Senecio gallicus, a diploid, annual plant that occurs in both coastal and ruderal inland areas of the Iberian Peninsula and southern France. The species appears to have a strong propensity for long-distance seed dispersal. Five cpDNA haplotypes were found by RFLP analysis among a sample of 111 individuals derived from 11 populations. Differences in haplotype frequencies across populations were most evident with respect to a dramatic increase in the frequency of a derived haplotype from coastal to inland localities. The level of cpDNA differentiation among populations within the inland group (θ 0 = 0.07) was significantly less than that seen within the coastal group (θ 0 = 0.41). In contrast, for allozymes, no significant difference in population structure was evident between collections from coastal and inland habitats. At the rangewide geographic scale, there was only a very weak association between inferred levels of gene flow and geographic distance for cpDNA, and no such association was found for allozymes. It appears that while seed movement in the species might be sufficiently great to disturb the pattern of isolation by distance for cpDNA, it cannot fully account for the nearly randomized spatial structure at polymorphic allozyme loci. It is suggested that isolation of populations in Atlantic-Mediterranean coastal refugia during previous glacial maxima, and the effects of subsequent colonization events in inland areas, have had an important effect on molding the present genetic structure of the species., (© 1998 The Society for the Study of Evolution.)

The sawflies that feed on the plant family Salicaceae can be divided into eight informal groups based on larval feeding habit or gall type: (1) species with free-living larvae; (2) leaf folders; (3) leaf blade gallers; (4) apical leaf gallers; (5) basal leaf gallers; (6) midrib and petiole gallers; (7) stem gallers; and (8) bud gallers. It has been proposed that the galling habit evolved from free-living larvae via leaf folders, and that the different gall types evolved gradually in the sequence mentioned above. Thus, the galling site would have "wandered" from the leaf margin toward the stem as a result of gradual changes in oviposition site preference. Allozyme data from eight informative loci were used to reconstruct the phylogeny of 18 representative sawfly species. The results suggest that indeed leaf folders seem to be a basal group; leaf blade gallers evolved independently of the other true gallers; apical and basal leaf gallers are not the ancestors of petiole and bud gallers, but they may share a common galling ancestor; bud gallers evolved from midrib/petiole gallers; and stem gallers are polyphyletic. The cause for the observed wandering of the galling site could be intraspecific competition due to a possible "nutrient shading effect" of galls situated closer to the host plant's main vascular system., (© 1998 The Society for the Study of Evolution.)

The phylogeography of the freshwater fish fauna of the southeastern United States has almost achieved paradigm status in evolutionary biology (Avise 1992), and the major geographic features responsible for shaping species distributions are well-characterized. Nevertheless, variation among species in distributions of allele or haplotype frequencies suggests that species-specific processes (e.g., migration) may also play a role in establishing those distributions. There has also been relatively little investigation into how population structure may differ among subregions in the Southeast, for example, on the Florida peninsula versus the U.S. mainland to the northwest and/or northeast. The geology of the peninsula is such that both physical and biotic fluctuations may have been (and still be) particularly important in establishing the population structure of freshwater taxa. This possibility leads to two interesting questions in population genetics. (1) Does gene flow in freshwater species of the region better approximate a one- or two-dimensional pattern? (2) Are populations on the peninsula farther from migration-genetic drift equilibrium than their counterparts on the mainland? These questions were addressed by examining the population strucuture of a livebearing fish, Heterandria formosa; several features of the biology of the species make it particularly likely that recent gene flow has been important in its evolution. I surveyed electrophoretic variation in 34 populations distributed throughout the species range. The phylogeographic patterns observed are in general concordance with those found in other species, although with some differences. A two-dimensional hypothesis of gene flow on the Florida peninsula better explains the data than does a one-dimensional one. There is no evidence that populations on the peninsula are farther from migration-drift equilibrium than those to the northwest. Populations in the northeast have lower genetic diversity than those to the south and west and show no isolation by distance; those results are consistent with a recent range expansion into the northeast, although smaller historical effective population sizes could also explain the pattern., (© 1998 The Society for the Study of Evolution.)

Both mtDNA variation and allozyme data demonstrate that geographic groupings of different color morphs of the starfish Linckia laevigata are congruent with a genetic discontinuity between the Indian and Pacific Oceans. Populations of L. laevigata sampled from Thailand and South Africa, where an orange color morph predominates, were surveyed using seven polymorphic enzyme loci and restriction fragment analysis of a portion of the mtDNA including the control region. Both allozyme and DNA data demonstrated that these populations were significantly genetically differentiated from each other and to a greater degree from 23 populations throughout the West Pacific Ocean, where a blue color morph is predominant. The genetic structure observed in L. laevigata is consistent with traditional ideas of a biogeographic boundary between the Indian and Pacific Oceans except that populations several hundreds kilometers off the coast of north Western Australia (Indian Ocean) were genetically similar to and had the same color morphs as Pacific populations. It is suggested that gene flow may have continued (possibly at a reduced rate) between these offshore reefs in Western Australia and the West Pacific during Pleistocene falls in sea level, but at the same time gene flow was restricted between these Western Australian populations and those in both Thailand and South Africa, possibly by upwellings. The molecular data in this study suggest that vicariant events have played an important role in shaping the broadscale genetic structure of L. laevigata. Additionally, greater genetic structure was observed among Indian Ocean populations than among Pacific Ocean populations, probably because there are fewer reefs and island archipelagos in the Indian Ocean than in the Pacific, and because present-day surface ocean currents do not facilitate long-distance dispersal., (© 1998 The Society for the Study of Evolution.)

Host plant-associated fitness trade-offs are central to models of sympatric speciation proposed for certain phytophagous insects. But empirical evidence for such trade-offs is scant, which has called into question the likelihood of nonallopatric speciation. Here, we report on the second in a series of studies testing for host-related selection on pupal life-history characteristics of apple- (Malus pumila L.) and hawthorn- (Crataegus mollis L. spp.) infesting races of the Tephritid fruit fly, Rhagoletis pomonella (Walsh). In particular, we examine the effects of winter length on the genetics of these flies. We have previously found that the earlier fruiting phenology of apple trees exposes apple-fly pupae to longer periods of warm weather preceding winter than hawthorn-fly pupae. Because R. pomonella has a facultative diapause, we hypothesized that this selects for pupae with more recalcitrant pupal diapauses (or slower metabolic/development rates) in the apple-fly race. A study in which we experimentally manipulated the length of the prewintering period for hawthorn-origin pupae supported this prediction. If the period preceding winter is important for apple- and hawthorn-fly pupae, then so too should be the length (duration) of winter; the rationale for this prediction is that "fast developing" pupae that break diapause too early will deplete their energy reserves and disproportionately die during long winters. To test this possibility, we chilled apple- and hawthorn-origin pupae collected from a field site near Grant, Michigan, in a refrigerator at 4°C for time periods ranging from one week to two years. Our a priori expectation was that longer periods of cold storage would select against allozyme markers that were associated with faster rates of development in our earlier study. Since these electromorphs are typically found at higher frequencies in hawthorn flies, extending the overwintering period should favor "apple-fly alleles" in both races. The results from this "overwinter" experiment supported the diapause hypothesis. The anticipated genetic response was observed in both apple and hawthorn races, as allele frequencies became significantly more "apple-fly-like" in eclosing adults surviving longer chilling periods. This indicates that it is the combination of environmental conditions before and during winter that selects on the host races. Many tests for trade-offs fail to adequately consider the interplay between insect development, host plant phenology, and local climatic conditions. Our findings suggest that such oversight may help to explain the paucity of reported fitness trade-offs., (© 1997 The Society for the Study of Evolution.)

Samples of Luxilus cornutus, Luxilus chrysocephalus, and their hybrids were collected along hypothesized routes of dispersal from Pleistocene refugia to examine the significance of geographic variation in patterns of introgression between these species. Patterns of allozyme and mitochondrial DNA (mtDNA) variation were generally consistent with those from previous studies. Tests of Hardy-Weinberg equilibrium revealed significant deficiencies of heterozygotes in all samples, indicating some form of reproductive isolation. Mitochondrial DNAs of each species were not equally represented in F 1 hybrids; however, this bias was eliminated when the two largest samples were excluded from the analysis. Backcross hybrids exhibited biased mtDNA introgression, as samples from Lake Erie (eastern) and Lake Michigan (western) drainages showed significant excesses of mtDNAs from L. chrysocephalus and L. cornutus, respectively, relative to frequencies of diagnostic allozyme markers. The extent and direction of allozyme and mtDNA introgression was quantified by calculating isolation index values from morphologically "pure" individuals of each species from each locality. Analysis of variance of these measures identified limited introgression of allozyme variants with no geographic pattern, but significant differences in direction of mtDNA introgression between drainages (i.e., postglacial dispersal route). Association between patterns of mtDNA introgression and dispersal route across the latitudinal width of the contact zone is best explained by genetic divergence during past isolation of ancestral populations from these drainages. These results identify a significant role for historical effects in the evolution of reproductive isolation and the process of speciation., (© 1997 The Society for the Study of Evolution.)

The hypothesis that levels of gene flow among populations are correlated with dispersal ability has typically been tested by comparing gene flow among species that differ in dispersal abilities, an approach that potentially confounds dispersal ability with other species-specific differences. In this study, we take advantage of geographic variation in the dispersal strategies of two wing-dimorphic planthopper species, Prokelisia marginata and P. dolus, to examine for the first time whether levels of gene flow among populations are correlated with intraspecific variation in dispersal ability. We found that in both of these coastal salt marsh-inhabiting species, population-genetic subdivision, as assessed using allozyme electrophoresis, parallels geographic variation in the proportion of flight-capable adults (macropters) in a population; in regions where levels of macroptery are high, population genetic subdivision is less than in regions where levels of macroptery are low. We found no evidence that geographic variation in dispersal capability influences the degree to which gene flow declines with distance in either species. Thus, both species provided evidence that intraspecific variation in dispersal strategies influences the genetic structure of populations, and that this effect is manifested in population-genetic structure at the scale of large, coastal regions, rather than in genetic isolation by distance within a region. This conclusion was supported by interspecific comparisons revealing that: (1) population-genetic structure (G ST ) of the two Prokelisia species correlated negatively with the mean proportion of flight-capable adults within a region; and (2) there was no evidence that the degree of isolation by distance increased with decreasing dispersal capability. Populations of the relatively sedentary P. dolus clustered by geographic region (using Nei's distances), but this was not the case for the more mobile P. marginata. Furthermore, gene flow among the two major regions we surveyed (Atlantic and Gulf Coasts) has been substantial in P. marginata, but relatively less in P. dolus. The results for P. marginata suggest that differences in the dispersal strategies of Atlantic and Gulf Coast populations occur despite extensive gene flow. We argue that gene flow is biased from Atlantic to Gulf Coast populations, indicating that selection favoring a reduction in flight capability must be intense along the Gulf. Together, the results of this study provide the first rigorous evidence of a negative relationship within a species between dispersal ability and the genetic structure of populations. Furthermore, regional variation in dispersal ability is apparently maintained by selective differences that outweigh high levels of gene flow among regions., (© 1997 The Society for the Study of Evolution.)

The Pacific island geckos Gehyra mutilata and Gehyra oceanica were studied on several Pacific Basin archipelagos to determine the degree that their distributions have been modified by humans (as commensals), through the analysis of protein variation using starch gel electrophoresis. Gehyra mutilata is an anthropophilic species that is widespread in the Pacific Basin and Southeast Asia. No protein variation was found in the Pacific Basin and southern Asia, although there were fixed allelic differences between populations of southern Asia and those further north. These results suggest possible recent human-aided transport across the Pacific from a population that experienced a genetic bottleneck in southern Asia. Gehyra oceanica, based on protein variation, consists of two natural groups in the Pacific, a northern (Micronesian) form and a southern (Melanesian and Polynesian) form. The northern form has very similar gene frequencies across its range in Micronesia. The southern form has its greatest allelic diversity in the south-central Pacific. F-statistics for G. oceanica in the south fall within the range of values in the literature for mainland Australian species of Gehyra that are not human commensals and for other island lizards that have been considered as natural dispersers. These values are consistent with the hypothesis that G. oceanica was naturally dispersed across the Pacific, prior to the arrival of humans and that the equatorial currents are a barrier to natural, north-south gene flow/dispersal in Pacific Basin lizards. However, human-aided dispersal within the northern and southern regions cannot be ruled out. By comparing the ecology of these two species, G. oceanica has the adaptations necessary for natural oversea dispersal, whereas G. mutilata has an ecology consistent with human-mediated dispersal, in support of the conclusions from the genetic data., (© 1997 The Society for the Study of Evolution.)

Meta-analyses of published correlation coefficients between multilocus heterozygosity (MLH) and two fitness surrogates, growth rate and fluctuating asymmetry, suggested that the strength of these correlations are generally weak. A variety of plants and animals was included in the meta-analyses. A statistically homogeneous group of MLH-growth rate correlation coefficients that included both plants and animals yielded a common correlation of r z = 0.133. A common correlation of r z = -0.170 was estimated for correlations between MLH and fluctuating asymmetry in three species of salmonid fishes. These results suggest that selection, including overdominance, has at most a weak effect at allozyme loci and cast some doubt on the widely held notion that heterozygosity and individual fitness are strongly correlated., (© 1996 The Society for the Study of Evolution.)

Phylogeography seeks to evaluate the relationship between genetic variation and geographic distribution of a species to examine the influence of historical events on divergence. Congruent phylogeographic patterns in codistributed species indicate historical association of the taxa being compared, and the uniform action of biogeographic events in shaping genetic variation. We sought to evaluate the congruence of patterns of genetic variation of five closely related fish species across a well-defined biogeographic boundary. We gathered allozymic and mitochondrial DNA (mtDNA) sequence data for five species of darters (Percidae: Etheostomatini) from populations distributed among biogeographic regions in the Ozark and Ouachita highlands of the south-central United States. Comparisons among species revealed noncongruence in the magnitude of genetic divergence in both allozymes and mtDNA sequences. We hypothesized that noncongruence resulted, in part, from differences in life histories of the species in our comparison. To address this hypothesis, we evaluated the association between gene flow (measured by N e m) and variation in body size and fecundity variables because they have been shown to influence gene flow in fishes. Correlation analysis revealed an association between gene flow and fecundity (r = 0.88), but not with body size (r = 0.36) or reproductive investment per individual (r = -0.23). The result was similar when independent contrasts of the original variables were used in correlation analyses. Phylogeographic analysis of mtDNA sequence data indicated the importance of history, evident in gene trees of Percina nasuta and Percina phoxocephala. Divergence rates between these two taxa may differ because of historically persistent differences in population sizes, reflected in present-day abundance and fecundity differences. Conversely, Percina caprodes showed little evidence of divergence in mtDNA sequences and yielded the highest mean N e m values from allozyme data. Comparisons among closely related, codistributed taxa may help discriminate among the possibilities for noncongruence in biogeographic studies by allowing evaluation of the influence of life history on patterns of gene flow., (© 1996 The Society for the Study of Evolution.)

Life cycle and breeding system variation in Epichloë grass endophytes (choke disease) is tightly linked to the degree of stroma formation. It is not known whether this variation results from differences in host resistance, fungal virulence, or environmental conditions. We found genetic differentiation between 173 asymptomatic (NS) and 93 stromata-forming (S) Epichloë strains isolated from one grass species, Brachypodium sylvaticum, based on 13 presumed allozyme loci, of which six were variable. The fungal strains originated from 10 sites in Switzerland, three sites of which were represented by both NS and S subpopulations. In total, 19 allozyme genotypes, that were nonrandomly distributed among S and NS were detected. Genetic variation measured as G ST between S and NS strains isolated from the same site ranged from 0.73 to 0.98. Clonality, measured as linkage disequilibrium at one site, was significant in the NS subpopulation (P ≪ 0.001), but not in the S subpopulation (P = 0.21), implying asexual reproduction by NS strains as well as successful horizontal transmission of S strains. Since all seeds are usually infected vegetatively, horizontal transmission implies the occurrence of multiple host infections. Altogether, these results provide indirect evidence that NS and S strains do not belong to one panmictic population and that differentiation patterns of stroma formation found in nature are due to genetic differences among fungi in associations with their host plants. We discuss the direction of evolution of disease expression in this system. The distribution of genetic variability suggests that the asymptomatic strains were derived from stromata-forming populations., (© 1996 The Society for the Study of Evolution.)

The relationship between gene flow and geographic proximity has been assessed for many insect species, but dispersal distances are poorly known for most of these. Thus, we are able to assess the concordance between vagility and gene flow for only a few species. In this study, I documented variation at six allozyme loci among Washington and Oregon populations of the sedentary, patchily distributed, lycaenid butterfly, Euphilotes enoptes (Boisduval) to assess whether the relationship between gene flow and geographic distance is consistent with the dispersal biology of this species. Both a phenogram based on genetic distances between populations and a regression analysis of gene flow estimates on geographic distances showed a pattern consistent with genetic isolation by distance. Many estimates of gene flow among pairs of populations separated by more than 100 km exceeded the equivalent of 10 individuals exchanged per generation, a value much greater than would be predicted from the limited dispersal ability of this species. However, based on the allozyme data, genetic neighborhood size was estimated to be approximately 39 individuals, a value that is consistent with poor vagility. The results of this study speak to the power of stepping-stone gene flow among populations and are compared to the results of other studies that have examined the relationship between dispersal and gene flow in sedentary insects., (© 1996 The Society for the Study of Evolution.)

Many investigators categorize individuals from hybrid zones to facilitate comparisons among genotypic classes (e.g., parental, F 1 , backcross) for comparative studies in which components of fitness or geographic variation are being analyzed. Frequently, multiple character sets representing genetically independent traits are used to classify these individuals and various methodologies are employed to combine the classifications obtained from the different character sets. We adapted the principles of total evidence and taxonomic congruence (two formalized approaches used by systematists in formulating phylogenetic hypotheses) to address the problem of discriminating hybridizing species and classifying individuals from hybrid zones. As our model, we used two morphological (coloration and morphometric) and two molecular (allozyme and mitochondrial DNA restriction-fragment-length polymorphism) character sets that differentiate two stone crab species (Menippe adina and M. mercenaria). Using principal-components analysis, we determined that combining character sets and eliminating characters or character sets that did not have large eigenvector coefficients for the principal component that best separated the two species yielded the highest level of discrimination between species and allowed us to classify a broad range of morpho-genotypes as hybrids. For the stone crabs, three diagnostic allozyme loci and five diagnostic coloration characters best separated the species. The two character sets were not completely congruent, but they agreed in their classification of 50% of the individuals from the hybrid zone and rarely strongly disagreed in their classifications. Classification discrepancies between the two character sets probably represent variation between traits in interspecific gene flow rather than intraspecific, ecologically mediated variation. Our results support the assertions of previous investigators who espoused the benefits associated with using multiple character sets to classify individuals from hybrid zones and demonstrate that, if character sets are reasonably congruent and numerically balanced, combining diagnostic characters from multiple character sets (a total-evidence approach) can enhance discriminatory power between species and facilitate the assignment of hybrid-zone individuals to genotypic classes. On the contrary, classifying hybrid-zone individuals using character sets separately (a taxonomic-congruence approach) provides the opportunity to compare levels of introgression between species and to assess reasons for discordance among the data sets., (© 1996 The Society for the Study of Evolution.)