Properties of allelic variants of phosphoglucomutase from the sea anemone Metridium senile

The phosphoglucomutase (Pgm) locus from populations of the sea anemone Metridium senile has three alleles in natural populations from the northeastern coast of North America. Two of the alleles exhibit clinal variation north of Cape Cod, suggesting a possible association of allele frequency with environmental temperature. This clinal pattern is reproducible and stable over at least brief periods of time. The allozymes encoded by each of the six Pgm genotypes have been partially purified and characterized. The symmetrical pH optimum for Vmax is pH 7.5; the apparent Km (Kmapp) of glucose-1-phosphate declines monotonically as the pH increases from 6.5 to 8.5. There are no pronounced differences in heat stabilities of PGM produced by various genotypes, nor are there significant differences in specific activities. There are no differences in the sensitivity of Vmax to temperature. Kmapp values are very low for all genotypes, ranging from about 2 to 12 microM, depending upon the temperature. Kmapp of glucose-1-phosphate declines as the temperature is raised for all genotypes, whether the pH is held constant or allowed to vary with the temperature. Under certain conditions, there are small significant differences among genotypes in Kappm values, but there is no systematic pattern to these differences. The present data provide no biochemical explanation for the maintenance of the Pgm cline by selection for functional differences under different thermal regimes.     

Isozyme diversity in sour,sweet, and ground cherry

Thirty-six sour (Prunus cerasus L.), sweet (P. avium L.), and ground cherry (P. fruticosa Pall.) selections were evaluated for seven enzyme systems and principal coordinate analysis was used to examine isozyme divergence among these cherry species. The enzyme systems studied were phosphoglucose isomerase (PGI), isocitrate dehydrogenase (IDH), phosphoglucomutase (PGM), 6-phosphogluconate dehydrogenase (6-PGD), leucine aminopeptidase (LAP), shikimate dehydrogenase (SKDH), and malate dehydrogenase (MDH). The first principal coordinate, which accounted for 41% of the total variation, separated the diploid sweet cherry selections from the sour, ground, and sour x ground cherry tetraploids. An additional 86 selections were evaluated for up to six of the enzyme systems to determine the polymorphisms at the enzyme loci and the level of heterozygosity between the diploid sweet cherry and the tetraploid species and interspecific hybrids. 6-PGD was the most polymorphic enzyme exhibiting 16 patterns. The tetraploid cherry species were more heterozygous than the diploid sweet cherry with an average heterozygosity of 78% compared to 19% for the diploids.     

ELECTROPHORETIC ANALYSIS OF GENETIC VARIATION IN THE CHAROPHYTA. I. GENE DUPLICATION VIA POLYPLOIDY1

Electrophoretic analysis of 12 species of Chara indicates that functional gene duplication via polyploidy has commonly occurred in the genus. Duplication has been followed by differentiation of the loci encoding phosphoglucose isomerase (PGI). This has led directly to generation of substantial levels of genie variation both within and between taxa. A simple genetic model is proposed to account for the large array of PGI phenotypes encountered in natural populations. Constraints imposed by the reproductive biology of members of the genus, such as selfing, appear to have resulted in selective premiums on intrinsic mechanisms of generating genetic variation. Levels of variation in PGI were higher than would be predicted on the basis of charophyte reproductive characteristics; haploid loci segregate approximately two alleles within each species.     

Temperature-dependent kinetic variation among phosphoglucose isomerase allozymes from the wing-polymorphic water strider, Limnoporus canaliculatus

Phosphoglucose isomerase (PGI) allozymes were isolated from the wing- polymorphic water strider, Limnoporus canaliculatus, and were characterized biochemically with respect to temperature-dependent kinetic and thermostability properties. At higher temperatures, the allozymes exhibited significant differences in Michaelis constant (Km) values for substrates of both the forward and reverse reaction directions. Results were consistent with expectations of adaptive kinetic differentiation based on the latitudinal variation of PGI allele frequencies. PGI genotypes also differed with regard to maximal velocity (Vmax)/Km ratios at higher temperatures. These differences were due primarily, if not exclusively, to allozyme-dependent variation in Km values. The allozymes also exhibited dramatic differences in thermostability. However, no thermostability differences were observed when the substrate analogue 6-phosphogluconate was present in the incubation medium. The data from this study, together with data from Mytilus edulis and Metridium senile on temperature-dependent kinetic variation among PGI allozymes, form a consistent picture of natural selection influencing the clinal variation of alleles at this locus in these three phylogenetically distant organisms. More definitive support of this hypothesis, however, must await additional studies on the physiological effects of the allozymic variation as well as direct measurements of fitness differences among the enzyme genotypes.      

Studies of esterase 6 in Drosophila melanogaster. I. The genetics of a posttranslational modification

A locus has been found, an allele of which causes a modification of some allozymes of the enzyme esterase 6 in Drosophila melanogaster. There are two alleles of this locus, one of which is dominant to the other and results in increased electrophoretic mobility of affected allozymes. The locus responsible has been mapped to 3-56.7 on the standard genetic map (Est-6 is at 3-36.8). Of 13 other enzyme systems analyzed, only leucine aminopeptidase is affected by the modifier locus. Neuraminidase incubations of homogenates altered the electrophoretic mobility of esterase 6 allozymes, but the mobility differences found are not large enough to conclude that esterase 6 is sialylated.This work was supported by NIH Grant No. GM23706 and PHS Grant SO7RR7031 to Rollin C. Richmond and by NIH Genetics Training Grant No. 82 to Indiana University.     

Phosphoglucose isomerase polymorphism in the common goby, Pomatoschistus microps (Krøyer) (TeleosteirGobiidae), around the British Isles

Phosphoglucose isomerase (PGI) polymorphism has been investigated by horizontal starch-gel electrophoresis in the common goby, Pomatoschistus microps (Krøyer), from 21 localities around the British Isles and at Roscoff, France. Sixteen PGI phenotypes were observed, consistent with occurrence of four alleles at each of the two loci (A,B). In the Teign estuary at Shaldon, Devon, seasonal fluctuation in PGI-A allele frequencies was accompanied in some months by departure from Hardy-Weinberg equilibrium. At this locality, a significant change in PGI-A allele frequencies was found with increasing modal age of sample (within a year class) but not between year-classes over eight years. Similar stability is noted for other areas. Major alleles (PGI-A2, A3; PGI-B2, B3) were found at all localities, but with marked differences in proportions. Atlantic coast populations tended to have higher A2 and B2 frequencies than those of the North Sea and English Channel, with Irish Sea populations having high A2 but low B2. Historical and selective causes for this geographical variation are discussed. A significant correlation exists between the frequencies of the major PGI-A alleles and maximum monthly mean sea temperature for the same area.     

Biochemical genetics of Fundulus heteroclitus (L.). IV. Spatial variation in gene frequencies of Idh-A,Idh-B, 6-Pgdh-A,and Est-S

The spatial variation in gene frequencies of four unlinked polymorphic loci was studied in the teleost Fundulus heteroclitus. Three loci (Idh-A, Idh-B, and Est-S) exhibit significant north-south clinal variation in allelic frequencies along the Atlantic Coast of North America, while a fourth locus (6-Pgdh-A) shows a modest clinal variation. These data, together with our previous data for Ldh-B, Mdh-A, Gpi-B, and Pgm-A, reveal a pattern of low gene diversity in the colder northern extremes of the species range and high gene diversity in warmer southern latitudes.This work was supported by Grants DEB76-19877 and DEB79-12216 from the National Science Foundation and by Grant P60-80-04 from the State of Maryland. REC and RVB were supported by NIH Training Grant GM07231 to the Department of Biology.Contribution No. 1104 from the Department of Biology, The Johns Hopkins University.     

Inheritance,intracellular localization,and genetic variation of phosphoglucomutase isozymes in maize (Zea mays L.)

Phosphoglucomutase (PGM; EC 2.7.5.1) isozyme variants were studied in a large number of inbred lines, crosses, and races of maize (Zea mays L.). Patterns of Mendelian inheritance demonstrated for PGM isozyme variants indicated that they are encoded by nuclear genes. Two unlinked loci, Pgm1 and Pgm2, located on the long arm of chromosome 1 and the short arm of chromosome 5, respectively, specify the observed electrophoretic variation on starch gels. No intra- or interlocus hybrid bands were found, suggesting that each isozyme band consists of a single polypeptide. PGM isozymes were present in all plant parts studied and the activity specified by both loci appears to reside in the cytoplasm. In studies of 520 racial collections of maize from Latin America, a single allele at each locus predominated in most collections. Likewise, the same alleles predominated in a set of 406 inbred lines of maize from the United States and Canada.This work was supported in part by NIH Research Grant GM 11546.Paper No. 8496 of the Journal Series of the North Carolina Agricultural Research Service, Raleigh, North Carolina.     

Adaptation at Specific Loci. IV. Differential Mating Success among Glycolytic Allozyme Genotypes of Colias Butterflies

Male mating success as a function of genotype is an important fitness component. It can be studied in wild populations, in species for which a given group of progeny has exactly one father, by determining genotypes of wildcaught mothers and of sufficient numbers of their progeny. Here, we study male mating success as a function of allozyme genotype at two glycolytic loci in Colias butterflies, in which sperm precedence is complete, so that the most recent male to mate fathers all of a female's subsequent progeny.--For the phosphoglucose isomerase, PGI, polymorphism, we predict mating advantage and disadvantage of male genotypes based on evaluation of their biochemical functional differences in the context of thermal-physiological-ecological constraints on the insects' flight activity. As predicted, we find major, significant advantage in mating success for kinetically favored genotypes, compared to the genotype distribution of males active with the sampled females in the wild. These effects are repeatable among samples and on different semispecies' genetic backgrounds.--Initial study of the phosphoglucomutase, PGM, polymorphism in the same samples reveals heterozygote advantage in male-mating success, compared to males active with the females sampled. This contrasts with a lack of correspondence between PGI and PGM genotypes in other fitness index or component differences.--Epistatic interactions in mating success between the two loci are absent.--There is no evidence for segregation distortion associated with the alleles of either primary locus studied, nor is there significant assortative mating.--These results extend our understanding of the specific variation studied and suggest that even loci closely related in function may have distinctive experience of evolutionary forces. Implications of the specificity of the effects seen are briefly discussed.     

Description and genetics of glucose phosphate isomerase (GPI) and phosphoglucomutase (PGM) polymorphisms in Asellus aquaticus (L.)

Analysis of Western European populations of Asellus aquaticus uncovered 10 electrophoretic phenotypes of glucose phosphate isomerase (GPI) and 7 of phosphoglucomutase (PGM). Breeding studies indicate that the variation is controlled by codominant alleles at two autosomal loci. Genotype frequencies in the two sexes do not differ significantly, mating between genotypes is random, and no structural linkage is detectable between the two loci. PGM shows nongenetic, "secondary" banding, particularly in animals stored at -20 degrees C prior to electrophoresis. This secondary banding confounds the identification of the genetic variation but can be controlled by the reducing agent 2-mercaptoethanol.     

Linkage relationships among five enzyme-coding gene loci in the copepod Tigriopus californicus: A genetic confirmation of achiasmatic meiosis

Linkage relationships among five polymorphic enzyme-coding gene loci in the marine copepod Tigriopus californicus have been determined using electrophoretic analysis of progeny from laboratory matings. Phosphoglucose isomerase (PGI; EC 5.3.1.9) was found to be tightly linked to glutamate-pyruvate transaminase (GPT; EC 2.6.1.2), with only one recombinant observed in 364 progeny; glutamate-oxaloacetate transaminase (GOT; EC 2.6.1.1) is linked to the PGI-GPT pair, with a recombination fraction of approximately 0.20 in male double heterozygotes. Phosphoglucomutase (PGM; EC 2.7.5.1) and an esterase (EST; EC 3.1.1.1) are not linked to the PGI, GPT, GOT grouping, which has been designated linkage group I. Reciprocal crosses have revealed that no recombination occurs in female T. californicus; this observation confirms a previous report that meiosis in female Tigriopus is achiasmatic.     

Purification and characterization of phosphoglucose isomerase allozymes from Daphnia magna.

Phosphoglucose isomerase (PGI, EC 5.3.1.9) is polymorphic in many populations. Frequently, it has been shown that naturally occurring allozymes exhibit strong deviations form Hardy-Weinberg expectations, suggesting fitness relevant mutations. To investigate the nature of this allozymic variation, PGI was purified from Daphnia magna to high purity yielding a specific activity of 135.2 U/mg. The kinetic parameters of the allozymes were characterized depending upon ionic strength, pH and viscosity. The half-saturation constants of the allozymes were all equal, while the specific activity of the PGI from heterozygotes was consistently higher than the PGI of the homozygotes, independent of pH, ionic strength and viscosity of the solution.     

Adaptation at Specific Loci. V. Metabolically Adjacent Enzyme Loci May Have Very Distinct Experiences of Selective Pressures

The polymorphic phosphoglucomutase (PGM) and glucose-6-phosphate dehydrogenase (G6PD) loci have been studied in parallel to experimental work on the phosphoglucose isomerase (PGI) polymorphism in Colias butterflies. PGI, PGM and G6PD are also autosomal in Colias. PGM and G6PD are loosely linked (and represent the first identified autosomal linkage group in Colias); they assort independently from PGI. Recombination occurs in both sexes. Neither PGM nor G6PD shows large, consistent differences in flight capacity through the day among its genotypes, as PGI does. PGM shows some change of allele frequencies, and match to Hardy-Weinberg expectation, with air temperature in middle and latter parts of the season, but not early in the season. G6PD may show some heterozygote excess over Hardy-Weinberg expectation early in the day, but more testing is needed. No evidence for differential survivorship was seen at PGM or G6PD, in contrast to PGI. At the PGM and G6PD loci, male heterozygotes are advantaged in mating with females, but without the evidence of female choice which occurs for PGI. These effects are not correlated among the three loci. There is no assortative mating at G6PD (nor at PGI). There is minor positive assortative mating of PGM heterozygotes, but it is too weak to account for the PGM-genotype-specific male mating advantage. No trends of multilocus genotype frequencies involving PGI are seen. Certain PGM-G6PD two-locus genotypes are over-represented, and others under-represented, in wild adult samples, particularly among males and uniformly among successfully mating males. Our results emphasize that enzyme loci sharing a substrate need not have common experience of the existence or strength of natural selection, and suggest initial food-resource processing and allocation as a possible context for fitness-related effects of the PGM and G6PD polymorphisms.     

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.     

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.     

Clinal genetic variation and the 'rare allele phenomenon' in random mating populations of <Emphasis Type="Italic">Urophora cardui</Emphasis> (Diptera: Tephritidae)

In the present study we investigate a contact zone between two population groups of the tephritid fly Urophora cardui. We investigate scenarios that may have produced the genetic differentiation of the two groups, and we describe the 'rare allele phenomenon' from the contact zone. The rare allele phenomenon refers to alleles that are found at high frequency in contact zones but are rare or lacking outside the contact zone. The phenomenon is often observed in hybrid zones between subspecies of limited reproductive compatibility, but seldom in populations with random mating. Clinal genetic variation was observed at three loci in the contact zone. Three alleles at the locus Aat showed steep clines, between 20--70 km wide. A rare Aat-A allele occurred at high frequency in the centre of the contact zone. Two further loci, Hk and Pgd, showed less steep clinal genetic variation, the transition being in and slightly south of the centre of the Aat cline. Populations showed Hardy--Weinberg proportions and there was no evidence for linkage dis-equlibrium. These findings suggest random mating and gradual introgression between the population systems, which may originate from at least two range expansions. Aat's steep clines and rare allele may indicate selection on Aat alleles, although we presently can not quantify any agents. Because U. cardui experiences random mating in the contact zone with no apparent 'hybrid' incompatibility, mating experiments offer the possibility for future enquiries about the genetic basis of the rare allele phenomenon.     

Allozyme Variation in a Threatened Freshwater Fish, Spotted Murrel (Channa punctatus) in a South Indian River System

Samples of the spotted murrel (Channa punctatus) were collected from three rivers of Tamil Nadu and Kerala. The allozyme variation of C. punctatus was investigated by polyacrylamide gel electrophoresis. Eighteen enzymes were detected, but only 10 (EST, PGM, G3PDH, G6PDH, SOD, GPI, ODH, GDH, XDH, and CK) showed consistent phenotypic variations. Allele frequencies were estimated at the 18 polymorphic loci representing 10 enzymes. Two rare alleles, EST-4*C and G6PDH-2*C, were noted in the Tamirabarani and Kallada populations but were absent in the Siruvani population. The allele frequencies of the Tamirabarani and Kallada populations were similar, except for a few loci. Among the three populations, the maximum genetic distance (0.026) and FST (0.203) were found between the geographically distant Siruvani and Kallada populations. Overall the study showed that among the three populations, the Tamirabarani and Kallada have similar genetic structures.     

Phosphoglucose isomerase gene duplication in the bony fishes: An evolutionary history

Electrophoretic patterns of phosphoglucose isomerase (PGI) in bony fishes provide strong evidence for a model of genetic control by two independent structural gene loci, most likely resulting from a gene duplication. This model is confirmed by a comparison of certain kinetic and molecular properties of the PGI homodimers (PGI-1 and PGI-2) isolated from extracts of the teleost Astyanax mexicanus. In addition, in most higher teleosts examined, the PGI enzymes show a regular pattern of tissue distribution, with PGI-2 predominant in muscle, the heterodimer often strongest in the heart, and PGI-1 predominant in liver and other organs. An examination of 53 species of bony fishes belonging to 38 families indicates a widespread occurrence of duplicate PGI loci and an early origin of the gene duplication, perhaps in the Leptolepiformes. The apparent presence of three PGI loci in trout and goldfish exemplifies how new loci can be incorporated into the genome through polyploidization.This research was supported in part by a NSF graduate traineeship to J.C.A., by the Clayton Foundation for Research in Biochemistry (G.B.K.), by NSF Grant GB-15644 and NIH Grant GM-15769 to Robert K. Selander, and by contract AT(38-1)-310 between the University of Georgia and the U.S. Atomic Energy Commission.     

THE ECOLOGICAL GENETICS OF INTRODUCED POPULATIONS OF THE GIANT TOAD,BUFO MARINUS. III. GEOGRAPHICAL PATTERNS OF VARIATION

The allele frequencies at ten polymorphic loci are described from 31 Bufo marinus populations in the Moreton Bay region in southeastern Queensland, Australia and the variation of these is found to be non-random in all cases. The pattern of non-randomness varies among loci, being clinal in two instances. The allele frequencies at the same ten loci are also described for 12 populations sampled from throughout B. marinus' Australian range. The frequency variation on this larger geographical scale is non-random at all but two loci (Mpi and Hbdh) and also varies among loci, in this case being clinal in four instances. In both cases, the patterns of variation are most reasonably explained as having resulted from genetic drift occurring during the recent range expansion which B. marinus is known to have experienced in Australia. It seems that natural selection has played little, if any, role in generating the observed gene frequency patterns. These results emphasize the need for caution in interpreting geographical patterns of variation. They show that even when clinal patterns exist at some loci but not at others, one cannot conclude that the patterns result from natural selection, unless the demographic histories of the studied populations are known and are inconsistent with the alternative hypothesis that the patterns result from genetic drift.     

Genetics of Lupinus

Summary Preliminary surveys indicate that most populations of Lupinus succulentus are genetically monomorphic for three flower color loci, viz., BB PP DD. In one small geographic area, a number of populations were polymorphic for the D/d locus. In this case, clinal variation and seasonal variation were found. The S/s locus, affecting seed coat pattern, was polymorphic in a large majority of populations and genetic variation at this locus is present in all but a very few colonies. When subdivisions of five large populations were studied, genetic differentiation was found within two of these populations. Gene frequencies tended toward the limits of zero or one in populations from relatively undisturbed sites but tended toward intermediate values in the more disturbed ruderal populations presumed to be recently colonized. It is concluded that recently colonized populations are not always genetically uniform.This work supported in part by National Institutes of Health Grant GM 10476