The functional impact of Pgm amino acid polymorphism on glycogen content in Drosophila melanogaster

Earlier studies of the common PGM allozymes in Drosophila melanogaster reported no in vitro activity differences. However, our study of nucleotide variation observed that PGM allozymes are a heterogeneous mixture of amino acid polymorphisms. In this study, we analyze 10 PGM protein haplotypes with respect to PGM activity, thermostability, and adult glycogen content. We find a twofold difference in activity among PGM protein haplotypes that is associated with a threefold difference in glycogen content. The latitudinal clines for several Pgm amino acid polymorphisms show that high PGM activity, and apparently higher flux to glycogen synthesis, parallel the low activity clines at G6PD for reduced pentose shunt flux in northern latitudes. This suggests that amino acid polymorphism is under selection at this branch point and may be favored for increased metabolic storage associated with stress resistance and adaptation to temperate regions.     

Extensive amino acid polymorphism at the pgm locus is consistent with adaptive protein evolution in Drosophila melanogaster

PGM plays a central role in the glycolytic pathway at the branch point leading to glycogen metabolism and is highly polymorphic in allozyme studies of many species. We have characterized the nucleotide diversity across the Pgm gene in Drosophila melanogaster and D. simulans to investigate the role that protein polymorphism plays at this crucial metabolic branch point shared with several other enzymes. Although D. melanogaster and D. simulans share common allozyme mobility alleles, we find these allozymes are the result of many different amino acid changes at the nucleotide level. In addition, specific allozyme classes within species contain several amino acid changes, which may explain the absence of latitudinal clines for PGM allozyme alleles, the lack of association of PGM allozymes with the cosmopolitan In(3L)P inversion, and the failure to detect differences between PGM allozymes in functional studies. We find a significant excess of amino acid polymorphisms within D. melanogaster when compared to the complete absence of fixed replacements with D. simulans. There is also strong linkage disequilibrium across the 2354 bp of the Pgm locus, which may be explained by a specific amino acid haplotype that is high in frequency yet contains an excess of singleton polymorphisms. Like G6pd, Pgm shows strong evidence for a branch point enzyme that exhibits adaptive protein evolution.     

The structure of morphometric and allozyme variation in relict populations of Gypsophila fastigiata (Caryophyllaceae) in Sweden

Patterns of variation and the structuring of diversity in seed phenotype and allozymes were investigated in Sweden Gypsophila fastigiata, a perennial herb with a disjunct ‘Late Glacial relict’ distribution in eastern and northern Europe. The overall patterns of variation in allozymes and seed morphology are congruent and are significantly correlated with geographic distance. However, most of the congruence between the distance matrices based on allozymes and seed morphology is attributable to regional differentiation between the isolated Öland and Dalarna metapopulations. On a local scale, within the two metapopulations, seed shape variation is partially correlated with geography whereas allozyme variation is unrelated to the geographic disposition of the sampling sites. Despite the fact that regional metapopulations can be discriminated on the basis of seed shape and allozyme frequencies, relatively little of the total diversity in seed morphology and allozymes is due to differentiation between regions. Partitioning of diversity into its within- and among-regional, site and individual components, showed that the majority of diversity (52% for seed shape and 91% for allozymes) is stored within sites. Seventy-three percent of the variation in allozymes is due to variation within individuals (i.e. heterozygosity) whereas phenotypic variation in seed shape within individuals is low (0.4% of the total diversity). The events that led to the large scale disjunction of the Late Glacial distribution of G. fastigiata may have shaped the pattern of differentiation observed among the regional isolates of the species. However, the species' history of local population disjunction during post-glacial and historic times has not had a substantial impact on the spatial organization of allozyme and morphometric diversity within regions. Extensive local gene flow may have allowed the regional metapopulations to have functioned as extended effective populations–on a time scale of tens or hundreds of years–and may have hindered the loss of within-site diversity.     

Evolutionary implications of allozyme and RAPD variation in diploid populations of dioecious buffalograss Buchloë dactyloides

Buffalograss, Buchloë dactyloides, is widely distributed throughout the Great Plains of North America, where it is an important species for rangeland forage and soil conservation. The species consists of two widespread polyploid races, with narrowly endemic diploid populations known from two regions: central Mexico and Gulf Coast Texas. We describe and compare the patterns of allozyme and RAPD variation in the two diploid races, using a set of 48 individuals from Texas and Mexico (four population samples of 12 individuals each). Twelve of 22 allozyme loci were polymorphic, exhibiting 35 alleles, while seven 10-mer RAPD primers revealed 98 polymorphic bands. Strong regional differences were detected in the extent of allozyme polymorphism: Mexican populations exhibited more internal gene diversity (H_e= 0.20, 0.19) than did the Texan populations (H_e= 0.08, 0.06), although the number of RAPD bands in Texas (n= 62) was only marginally smaller than in Mexico (n= 68). F-statistics for the allozyme data, averaged over loci, revealed strong regional differentiation (mean F_RT=+ 0.30), as well as some differentiation among populations within regions (mean F_PR=+ 0.09). In order to describe and compare the partitioning of genetic variation for multiple allozyme and RAPD loci, we performed an Analysis of Molecular Variance (AMOVA). AMOVA for both allozyme and RAPD data revealed similar qualitative patterns: large regional differences and smaller (but significant) population differences within regions. RAPDs revealed greater variation among regions (58.4% of total variance) than allozymes (45.2%), but less variation among individuals within populations (31.9% for RAPDs vs. 45.2% for allozymes); the proportion of genetic variance among populations within regions was similar (9.7% for RAPDs vs. 9.6% for allozymes). Despite this large-scale concordance of allozyme and RAPD variation patterns, multiple correlation Mantel techniques revealed that the correlations were low on an individual by individual basis. Our findings of strong regional differences among the diploid races will facilitate further study of polyploid evolution in buffalograss.     

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

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

Genetic variation within and between populations of Potamogeton pusillus agg.

Patterns of isozyme variation were examined in 17 populations of P. pusillus and P. berchtoldii, together with one population of P. trichoides taken for comparison. Both P. pusillus and P. berchtoldii displayed low levels of variation within populations associated with high levels of interpopulation differentiation. This pattern of partitioning of genetic variation within and between populations is attributed to the founder effect, frequent vegetative propagation by turions, dominant self-fertilization and limited seedling recruitment. The mechanism of pollen transfer was investigated in cultivation. Effective pollination takes place in air above the water surface (autogamy, geitonogamy, anemogamy), on the water surface (epihydrogamy) or below water surface (hydroautogamy). The species are self-compatible. The low level of infra-population variation together with rare occurrence of heterozygotes suggest that selfing is the most frequent mode of pollination, although the protogynous flowers may occasionally permit some cross-pollination. Unique enzyme markers were found for P. pusillus and P. berchtoldii, and also for the single population of P. trichoides. All multienzyme phenotypes were species-specific. Isozyme data support the separate position of P. pusillus and P. berchtoldii. UPGMA dendogram based on enzyme data of 133 plant samples revealed three distinct main enzymatic entities perfectly corresponding to the three morphologically defined species.     

Inhibition of phosphoglucose isomerase allozymes from the wing polymorphic waterstrider,Limnoporus canaliculatus,by pentose shunt metabolites

Inhibition of phosphoglucose isomerase (PGI) allozymes from the wing-polymorphic waterstrider, Limnoporus canaliculatus, by three pentose-shunt metabolites was studied at several different temperatures. This was done to determine if the allozymes exhibited a differential ability to participate in lipid biosynthesis via differential partitioning of carbon flux through the pentose shunt versus glycolysis. 6-Phosphogluconate and erythrose-4-phosphate proved to be strong competitive inhibitors of PGI, while sedoheptulose-7-phosphate was a very weak inhibitor. The PGI allozymes from L. canalicualtus were differentially inhibited by 6-phosphogluconate at two of the three temperatures studied. However, this property does not appear to be an adaptive difference between the allozymes but, rather, a correlated effect resulting from variation in substrate binding. Estimates of reaction rates for the allozymes indicate that the differences in inhibition result in no detectable differences in reaction velocities. Thus, no evidence in support of the hypothesis that PGI allozymes from Limnoporus canaliculatus were adapted to function in different metabolic capacities via differential inhibition was obtained in this study. However, the importance of this characteristic in allozymic adaptation in natural populations remains an open question.Supported by NSF Grant DEB 7908802 and UPHS Grant GM 21133 to R. K. Koehn and an NSF dissertation improvement grant to A. J. Zera.     

Mating system and seed variation of <Emphasis Type="Italic">Acacia</Emphasis> hybrid (<Emphasis Type="Italic">A. mangium</Emphasis> × <Emphasis Type="Italic">A. auriculiformis</Emphasis>)

The mating system and seed variation of Acacia hybrid (A. mangium × A. auriculiformis) were studied using allozymes and random amplified polymorphic DNA (RAPD) markers, respectively. Multi-locus outcrossing rate estimations indicated that the hybrid was predominantly outcrossed (mean±s.e. t _m = 0.86±0.01). Seed variation was investigated using 35 polymorphic RAPD fragments. An analysis of molecular variance (AMOVA) revealed the highest genetic variation among seeds within a pod (66%–70%), followed by among pods within inflorescence (29%–37%), and the least variation among inflorescences within tree (<1%). In addition, two to four RAPD profiles could be detected among seeds within pod. Therefore, the results suggest that a maximum of four seeds per pod could be sampled for the establishment of a mapping population for further studies.     

Associations of allozyme variation and behavior in the cowpea weevil (Callosobruchus maculatus)

The cowpea weevil (Callosobruchus maculatus (Fabré)) exhibits several behavioral traits that are stable within, but vary among, strains. These traits are heritable and quantitative. We used cellulose acetate gel electrophoresis to quantify allozyme variation within and among laboratory cultures of four weevil strains and determine whether allozyme variation correlates with behavioral traits. Significant variation exists at 8 of 11 loci assayed and gene frequencies are significantly different among strains. The South Indian strain (SI) is most variable and measures of genetic distance set it apart from the other strains. It is also behaviorally unique. The Brazilian strain (BC) is most different from SI in allozyme diversity and behavioral phenotype, while two African strains (IITA, CAM) are intermediate in allozyme diversity and phenotype. These results are consistent with the known history of these strains and the differences in the allozymes parallel the differences in behavioral traits.     

Allozyme frequencies in western European populations of Asellus aquaticus (L.) Isopoda and their association with water pollution

A study was made of allozyme variation in western European populations of Asellus aquaticus. Two enzymes were examined glucose phosphate isomerase (CPI) and phosphoglucomutase (PGM).
The populations show significant latitudinal clincs for GPI and PGM with the GPI-2 and PGM-2 alleles increasing in more southerly latitudes. While GPI-2 frequencies in British populations are consistent with the overall clinc, considered separately they show a reverse cline. This appears to be the result of a strong association of GPI-2 frequencies with water pollution combined with the geographic distribution of the polluted sites.
The association of high GPI-2 frequencies with high levels of water pollution is present in two independent sets of populations in Britain, and is more specifically associated with low levels of environmental oxygen. This latter association also occurs independently in temporal studies made at three locations. However, an additional temporal association of high GPI-2 frequencies with high temperatures also exists, consistent with the observed latitudinal cline. A study of adult mortality by oxygen deprivation at different temperatures suggests that both temperature and oxygen concentrations may act selectively with the GPI 2-2 genotype to be favoured under certain conditions. In contrast to GPI, PGM allozymes do not appear to be associated with pollution related variables.
The findings of the study point to selection acting on the two loci examined, though most convincingly for GPI. However, it remains to be demonstrated that selection is acting directly on the loci, and if so, how it operates to produce the patterns of allozyme variation observed in nature.     

Genetic differentiation in Hippocrepis emerus (Leguminosae): allozyme and DNA fingerprint variation in disjunct Scandinavian populations

The structure of genetic variation in disjunct Scandinavian populations of Hippocrepis emerus was studied using allozymes and DNA fingerprinting. Variation in the three native regional populations in Scandinavia was compared with that in a recently introduced population in Sweden. In contrast to the recently introduced population, the native Scandinavian isolates of H. emerus showed high levels of allozyme fixation and low levels of DNA diversity. Variation in allozymes and at DNA fingerprint loci showed closely congruent patterns of geographic variation, with pronounced differentiation between the native Norwegian and Swedish isolates of the species. The structure of genetic variation in native Scandinavian H. emerus is interpreted in terms of historical population bottlenecks and founder events during the species' postglacial immigration into Scandinavia.     

Behavior of Cape fur seals (Arctocephalus pusillus pusillus) in relation to temporal variation in predation risk by white sharks (Carcharodon carcharias) around a seal rookery in False Bay,South Africa

The marked differences in predation risk posed by white sharks (Carcarodon carcarias) at island rookeries of Cape fur seals (Arctocephalus pusillus pusillus) offer a quasi‐experimental design within a natural system for exploring how prey adjust their behavior in response to temporal variation in predation risk. Here we compare movement of juvenile and adult Cape fur seals at a high risk (Seal Island) and low risk (Egg Island) rookery. We further compare juveniles and adults at Seal Island in low and high risk seasons and at low and high risk times of day within those seasons. Adult fur seals at Seal Island avoided traversing the zone of high white shark predation risk during the high risk period (0700–0959) in the season of high risk (winter), but not during the low risk season (summer). By contrast, adult fur seals at Egg Island showed no temporal discretion in either season. Unlike juvenile fur seals at Egg Island, juveniles at Seal Island adjusted their temporal movement patterns to more closely mimic adult seal movement patterns. This suggests that exposure to predators is the primary driver of temporal adjustments to movement by prey species commuting from a central place.     

Effects of various metabolites on two phosphoglucomutase allozyme activities from Drosophila melanogaster

The effects of various metabolites on the two most common phosphoglucomutase allozymes (PGM^A and PGM^B) in Drosophila melanogaster have been investigated in vitro. 2,3-Diphosphoglycerate (2,3DPG) inhibited PGM^A and PGM^B to the same degree in the presence of 25 µM glucose-1,6-diphosphate (G1, 6P₂). However a higher concentration of G1,6P₂ partially reversed the inhibition of PGM^A exerted by 2,3DPG, so that in the presence of 150 µM G1,6P₂ the inhibition of PGM^A was half that of PGM^B at pH 6.0. Glycerol-3-phosphate (G3P) had no significant effect at pH 7.4 but exerted an activating effect at pH 6.0 which was more pronounced in the case of PGM^B. ATP, citrate, and fructose-1, 6-diphosphate (F1,6P₂) inhibited both PGM^A and PGM^B. The differences found in vitro between these two allozymes can have a significant impact on in vivo function and, therefore, on the maintenance of PGM polymorphism in experimental populations of D. melanogaster studied in the laboratory.     

Male terminalia variation in the rainforest dwelling <Emphasis Type="Italic">Drosophila teissieri</Emphasis> contrasts with the sperm pattern and species stability

It is commonly recognized that speciation does not necessarily imply extensive variation between populations, and what the speciation process per se consists of still remains an unanswered question. We advocate here that the variation of male terminalia does not necessarily result in noticeable reproductive isolation. We report whether there is invariance or variance of traits central to sexual selection processes (i.e. male terminalia and sperm length) compared to traits which are generally assumed to vary more neutrally (i.e. allozymes) in the strictly Afrotropical forest-dwelling continental species Drosophila teissieri. Three geographic blocks can be recognized along the present range of the species. Our data suggest that the components of the species integrity do not obey the variance/invariance alternative consistently. Male terminalia and allozymes show extensive variation while sperm length distribution is strikingly similar between the geographic blocks. It is therefore inferred that sperm length might be one of the major targets of stabilizing selection. Finally, it is suggested that the striking fit between the extent of sperm heteromorphism (within male) and sperm polymorphism (between males) is instrumental in maintaining the species integrity.     

Genetic variation within and among North Atlantic and Baltic populations of the benthic alga Phycodrys rubens (Rhodophyta)

Genetic variation was examined within and among North Atlantic, North Sea and Baltic populations of the benthic red alga Phycodrys rubens using allozymes and random amplified polymorphic DNA (RAPD) markers. On western and eastern North Atlantic coasts distinct allozyme types were found, with the exception of western Newfoundland where East and West Atlantic types co-occur. Along the European coasts, two genetic groups were distinguished by fixed allelic differences: an outer oceanic group and a North Sea/Baltic group. The two genetic types co-occur in the Skagerrak and Kattegat region. Reproductive isolation between the two types is suggested by the lack of hybrids in the overlap zones, and they may therefore represent sibling species. Unexpectedly, an analysis of RAPD variation was unable to recover the two cryptic species identified using allozymes. Within-population RAPD variation was similar to or greater than between-population variation. The lack of structure in the RAPD data cannot be attributed solely to technical artefacts of the method but appears to reflect real biological variability. Within-population genomic polymorphisms caused by frequent mutational events are discussed, as are high amounts of genetic drift and possible disruptive selection brought about by stressed habitats. Finally, Baltic and extra-Baltic salinity ecotypes are known to exist in P. rubens. However, no correlation between ecotypic variation and allozyme groups was detected.     

Genetic distinctness of three widespread and morphologically variable species of Drupella (Gastropoda,Muricidae)

Corallivorous gastropods of the genus Drupella have caused considerable damage to corals at widely separated reefs in the Indo-Pacific. Morphological variability of Drupella species within and between areas has caused taxonomic confusion. To clarify the relationships, we examined allozyme variation at 16 gene loci in samples from Western Australia, Queensland and Japan. Within sites, the species D. cornus, D. rugosa and D. fragum were distinguishable individually by each of 9 to 11 loci, with average genetic identities of about 0.25. The differences extended across sites, whereas the conspecific genetic identities over distances up to 6000 km were 0.86 to 1.00, supporting the view that there are three widespread species of Drupella. Nevertheless, there is much variation within species for allozymes, size, shape and colour.     

Genetic Variation of Mantled Howler Monkeys (Alouatta palliata) from Costa Rica1

We examined genetic diversity of howler monkeys (Alouatta palliata) from Costa Rica. Blood samples of howler monkeys were collected at various locations in Costa Rica, and electrophoresis of total plasma proteins yielded no variation. We also conducted starch gel electrophoresis of red cell isozymes and did not find variation for any of the 14 loci analyzed (i.e., ACP, ADA, CA2, EST, GPI, IDH, LDH‐1, LDH‐2, MDH, PGD, PGM‐1, PGM‐2, SOD, and TPI). These findings were compared with the levels of genetic variation for A. seniculus and A. belzebul from one Brazilian population. Four of the 14 isozymes (ADA, GPI, PGD, and SOD) showed more than one allele for these species. Both A. seniculus and A. belzebul from Brazil showed similar levels of genetic variation. The potential causes of the low genetic variation in A. palliata from Costa Rica are discussed.     

Evolutionary genetics of Metridium senile. II. Geographic patterns of allozyme variation

Electrophoretic surveys have demonstrated that populations of the sea anemone Metridium senile along the northeast coast of the United States are polymorphic at four enzyme loci. Phosphoglucose isomerase (PGI) has two alleles in most populations, phosphoglucomutase (PGM) has three alleles, and two leucine aminopeptidase loci have two common alleles each. Phosphoglucose isomerase displays clinal variation and an apparent association with environmental temperature. Phosphoglucomutase shows clinal variation north of Cape Cod for two of the three alleles, while the two leucine aminopeptidase loci are not clinal. All loci show a great deal of variation in populations on Cape Cod, but there is no apparent systematic pattern to this variation. Temperature may be a selective agent in the maintenance of the PGI and PGM clines, although other possibilities cannot presently be completely excluded.Supported by Grant T-4 from the Health Research and Services Foundation, NSF DEB77-14442, NIH GM25809, and NIH GM28024.     

Range‐wide genetic homogeneity in the California sea mussel (Mytilus californianus): a comparison of allozymes,nuclear DNA markers,and mitochondrial DNA sequences

We tested for genetic differentiation among six populations of California sea mussels (Mytilus californianus) sampled across 4000 km of its geographical range by comparing patterns of variation at four independent types of genetic markers: allozymes, single‐copy nuclear DNA markers, and DNA sequences from the male and female mitochondrial genomes. Despite our extensive sampling and genotyping efforts, we detected no significant differences among localities and no signal of isolation by distance suggesting that M. californianus is genetically homogeneous throughout its range. This concordance differs from similar studies on other mytilids, especially in the role of postsettlement selection generating differences between exposed coastal and estuarine habitats. To assess if this homogeneity was due to M. californianus not inhabiting estuarine environments, we reviewed studies comparing allozymes with other classes of nuclear DNA markers. Although both types of markers gave broadly consistent results, there was a bias favouring studies in which allozymes were more divergent than DNA markers (nine to three) and a disproportionate number of these cases involved marine taxa (seven). Furthermore, allozymes were significantly more heterogeneous than DNA markers in three of the four studies that sampled coastal and estuarine habitats. We conclude that the genetic uniformity exhibited by M. californianus may result from a combination of extensive gene flow and the lack of exposure to strong selective gradients across its range.     

RIBOSOMAL DNA AND RAPD VARIATION IN THE RARE PLANT FAMILY LACTORIDACEAE

Lactoris fernandeziana, endemic to the island of Masatierra in the Juan Fernandez Archipelago, is the only living member of the primitive angiosperm family, Lactoridaceae. The species was surveyed for ribosomal DNA (rDNA) and RAPD (Random Amplified Polymorphic DNA) variation. Previous analyses of allozymes had revealed no variation within the species. Variation was found for length in the intergenic spacer and for restriction sites in the 18S–25S genes of rDNA, and for the presence of amplified bands using 16 primers. Different rDNA repeat lengths and restriction site variants were detected within individuals as well as within and among populations. The level of variation in RAPDs is low relative to other Juan Fernandez endemic species surveyed, and nearly all variants were restricted to single populations. The rDNA length variants were distributed throughout the island, whereas the rDNA restriction site variants and RAPD markers indicated minor genetic differences among the populations.