Genetic coadaptation in the chromosomal polymorphism of Drosophila subobscura II. Changes of gametic disequilibrium in experimental populations

Experimental populations were examined for temporal changes of gametic disequilibria between allozyme loci (Lap and Pept-1) and gene arrangements of the O chromosome of Drosophila subobscura (O _st and O ₃₊₄₊₇) under several environmental conditions. In the foundation of the experimental populations a genetic perturbation was carried out in order to test the relevance of the current hypotheses used to explain the allozyme-inversion associations observed in natural populations. Differential changes of gametic disequilibria were detected over generations under the different environmental conditions. Mere mechanical or stochastic factors cannot explain the results and natural selection is probably the major agent generating the detected gametic associations. The observations are interpreted as a proof of coadaptation of D. subobscura inversions.     

A profile of Drosophila species' enzymes assayed by electrophoresis. I. Number of alleles,heterozygosities, and linkage disequilibrium in glucose-metabolizing systems and some other enzymes

Seven isolated large populations of Drosophila belonging to five different species were examined by starch gel electrophoresis for allozyme variation. Six to eleven enzyme loci in the glucose-metabolizing system (group I) and six to eight enzyme loci (group II) which were not directly involved in the above-mentioned system were assayed. The parameters estimated were the average number of alleles per locus, allele frequencies, proportions of polymorphic loci, and average heterozygosity per population for group I and group II loci. The major finding is that genetic variability measured by allozyme variations is much higher for group II than for group I enzymes in terms of every parameter in all the populations. This is consistent with the earlier findings in D. ananassae by Gillespie and Kojima (1968). Linkage disequilibrium, a measure of genome integration, was computed between an enzyme locus and an inversion segment of the same chromosome. The preliminary analysis of this aspect of the study indicates that no substantial linkage disequilibrium builds up between the chromosomal segments unless the pair of segments is less than 10 centimorgan units apart.This study was supported by USPHS Grant GM-15769 and Atomic Energy Commission Contract AT-(40-1)-3681.NIH Trainee supported by Grant 5 TO 1 GM 00337.     

Analysis of inbreeding in a colonizing population of Drosophila subobscura

An analysis of the effects of inbreeding on the genetic structure of a colonizing population of Drosophila subobscura has been carried out. Species of Drosophila, particularly D. subobscura, may have lethal alleles associated with chromosomal inversions and our aim was to assess the extent to which the genome is balanced in this way. The frequencies of chromosomal inversions were compared between a large population and a set of 72 lines that were maintained by brother-sister mating for 10 generations. Fisher's matrix method was used to calculate the expected homozygosity in these inbred lines for 5 allozyme loci (Aph, Hk-1, Lap, Odh, and Pept-1) used as markers of large chromosomal segments. Furthermore, the expected rates of fixation corresponding to these allozyme loci were also calculated. The results show that the amount of homozygosis observed did not differ significantly from expectations (with the corresponding loss of lines as a consequence of the reduction in viability). However, two deviations from strict neutrality were observed: there was a heterozygote excess at the Lap locus, and the frequency of the O ₅ inversion (always associated with a lethal gene in colonizing populations) was higher than expected.     

Gametic equilibrium of four loci on chromosome 1

Summary The present study is the first to investigate gametic disequilibrium of loosely linked loci (recombination >22%) of known map distance in humans. The study population consisted of 302 healthy unrelated individuals of white mixed-European ancestry living in the Denver, Colorado area. Pairwise combinations of four loci on chromosome 1 (Rhesus, Duffy, phosphoglucomutase-1, and amylase-2) were examined for nonrandom association of alleles at different loci. Observed and expected values were compared with chi-square goodness-of-fit tests. There was no evidence of first-order interaction, i.e., gametic disequilibrium. From these findings it seems that gametic disequilibrium may not be important in the genetic organization of chromosome 1.     

Gene activity of polytene chromosomes in Drosophila species of the obscura group

The polytene chromosome puffing patterns of Drosophila guanche were established and compared with those of Drosophila subobscura. A total of 150 loci, active in some of the 17 developmental stages studied, were described and 23 of them were found to form the characteristic puffing pattern of D. guanche. Taking into account the number of puffs as well as the gene activity of each chromosome and the total gene activity, D. guanche seems to be less active than D. subobscura. Although both species show a degree of homology in their puffing patterns lower than that found for sibling species, the degree of homology is stronger than that between species belonging to the same group but to different subgroups. Thus, D. guanche and D. subobscura must be considered as phylogenetically closely related species, belonging to the same subgroup.     

Dynamics of Correlated Genetic Systems. V. Rates of Decay of Linkage Disequilibria in Experimental Populations of DROSOPHILA MELANOGASTER

The dynamic behavior of four-locus gametic frequency distributions was studied in five replicate cage populations of Drosophila melanogaster for up to 50 generations. The joint frequency distributions were resolved into gene frequencies and various disequilibrium measures. In addition, F statistics for marginal single-locus genotypic frequency distributions were followed through time. The gene frequency, disequilibrium and F statistics were obtained for four chromosome 3 enzyme marker loci [isocitrate dehydrogenase (3–27.1), esterase-6 (3–36.8), phosphoglucomutase (3–43.4) and esterase-C (3–49.0)]. The initial structure of the experimental populations featured random mating proportions, and two complementary gametic types with respect to the marker loci, thus assuring complete pairwise linkage disequilibrium among the markers.——The experimental results indicate: (1) the between-replicate variance in gene frequency varied substantially among loci, with isocitrate dehydrogenase showing the greatest between-replicate variance, and esterase-C the least. (2) The F statistics initially were strongly negative but decayed to the neighborhood of zero for all marker loci except esterase-C. The rate at which the F statistics approached zero varied among the marker loci, indicating substantial differences in the distribution of selective effects along the chromosome. The centromeric region, marked by esterase-C, shows the strongest selective effects. (3) The rate of decay of linkage disequilibrium was much faster than expected for pairs of neutral loci, averaging 1.82 times the neutral rate over all replicates and pairs of loci. This acceleration, which was observed for all six pairwise combinations of loci, was interpreted as resulting from the interaction between selection and recombination. Our experimental results are consistent with many investigations of linkage disequilibrium in natural populations of Drosophila melanogaster that show little or no disequilibrium among enzyme loci. (4) A fortuitous contamination of two cages revealed an apparent regulatory interaction between the migrant and nonmigrant chromosomes at the esterase-C locus. The migrant chromosomes were very rapidly absorbed into the recipient populations, despite this interaction. This result suggests that the dynamics of migration in populations may be phenomenologically richer than anticipated by simple theory.     

Genetic Diversity and Linkage Disequilibrium in Drosophila melanogaster with Different Rates of Development

We have examined eight enzyme polymorphisms in groups of Drosophila melanogaster flies with fast, intermediate and slow development. The allelic frequencies are similar in all three developmental classes, but the distribution of the genotypes among the classes is significantly heterogenous for the three loci on the second chromosome. When the total sample of 300 individuals is examined, significant gametic disequilibrium appears in 3 out of 13 pairs of genes located on the same chromosome and in 4 out of 15 pairs of genes located on different chromosomes. This 25% incidence of disequilibrium between pairs of genes is larger than previously observed in other natural populations (but similar to the incidence observed in laboratory populations). The greater frequency of significant gametic disequilibrium in our study is probably due to the larger number of genomes sampled.-Some models specifically predict that individuals with faster rates of development (i.e., greater fitness) should be more heterozygous (and exhibit more linkage disequilibrium) than individuals with slower development. This hypothesis is not supported by our results.     

A long-term study on seasonal changes of gametic disequilibrium between allozymes and inversions in Drosophila subobscura

Seasonal variation (spring, early summer, late summer, and autumn) of gametic disequilibrium between gene arrangements (OST and O3+4) of the O chromosome and Lap, Pept-1, and Acph allozyme loci, located inside these inversions, has been recorded in a natural population of Drosophila subobscura during seven years over a 15-year period. The length of the study allowed us to investigate the temporal variation of the allozyme-inversion associations by statistical methods of time series analysis. Cyclic seasonal changes of allozyme-inversion associations for both Lap and Pept-1 are detected in the natural population. In both cases, the patterns of seasonal change are due to the seasonal change of frequency of Lap and Pept-1 allozymes occurring exclusively within the OST gene arrangement. In contrast, the allozyme frequencies at these loci within the O3+4 gene arrangement are stable along seasons. The patterns of temporal variation of allozyme-inversion associations for Lap and Pept-1 in the natural population are contrasted with those previously published that correspond to gene arrangements of the O chromosome and nucleotide polymorphism at the rp49 region located inside these inversions, suggesting that natural selection is operating on these allozyme-inversion associations.     

The role of genic selection in the establishment of inversion polymorphism in Drosophila subobscura

A model for explaining the establishment of newly arisen inversions in natural populations, in which the inverted segment may be selected for if its load of deleterious mutations is smaller than the average load of the noninverted segment in the population, is tested for Drosophila subobscura. The results show that for new inversions, originally with no deleterious alleles, the expected cumulative distribution of inversion lengths fits fairly well with the observed one. Therefore, genic selection may be an important cause of the establishment of newly arisen inversions in natural populations of D. subobscura. The applicability of the model to the maintenance of the inversion polymorphism present in this species; is discussed.     

Multilocus Structure of Natural Populations of HORDEUM SPONTANEUM

The association of alleles among different loci was studied in natural populations of Hordeum spontaneum, the evolutionary progenitor of cultivated barley. The variance of the number of heterozygous loci in two randomly chosen gametes affords a useful measure of such association. The behavior of this statistic in several particular models is described. Generally, linkage (gametic phase) disequilibrium tends to increase the variance above the value expected under complete independence. This increase is greatest when disequilibria are such as to maximize the sum of squares of the two-locus gametic frequencies.—When data on several loci per individual are available, the observed variance may be tested for its agreement with that expected under the hypothesis of complete interlocus independence, using the sampling theory of this model. When applied to allozyme data from 26 polymorphic populations of wild barley, this test demonstrated the presence of geographically widespread multilocus organization. On average, the variance was 80% higher than expected under random association. Gametic frequencies for four esterase loci in both of these populations of wild barley and two composite crosses of cultivated barley were analyzed. Most generations of the composites showed less multilocus structure, as measured by the indices of association, than the wild populations.     

Allozyme polymorphism in <Emphasis Type="Italic">Drosophila</Emphasis>

Every population possesses genetic variations which are achieved through gene mutation, genetic recombination, hybridization, gene duplication etc. These genetic variations provide raw materials for evolutionary forces to create a better surviving species. Genetic polymorphism is reflected at every level in the populations, for example, at phenotypic, chromosomal, protein and DNA levels. Protein or enzyme polymorphisms have been well studied in various organisms including Drosophila and humans. Drosophila has proven to be a good model organism for carrying out polymorphism studies. Among the different species of Drosophila, there is a wide variation in the levels of allozyme polymorphisms and heterozygosities which depends upon species, geographical regions, number and nature of loci in question etc. In Drosophila, the average polymorphic enzyme loci and average heterozygosity ranges from 35 to 70 percent and 10 to 20 percent respectively. The genetic differentiation as observed through allozyme or isozyme variation affords an important parameter in evaluating the phylogenetic relationships between different species of Drosophila and also for discussing the adaptive significance of allozyme polymorphisms. Therefore, this review attempts to compile all studies on allozyme polymorphism in Drosophila that have been undertaken so far.     

Sample sizes required to detect linkage disequilibrium between two or three loci.

Estimates of the degree of nonrandom association among genes (linkage disequilibrium) can provide evidence of the role of natural selection in maintaining allozyme polymorphisms in natural populations. This paper outlines the maximum likelihood procedures for such estimates based on gametic or zygotic frequencies at the level of two loci. The analysis is extended to estimating disequilibrium between three loci. In particular, the question of the sampling requirements to detect different intensities of disequilibrium is considered. It is found that relatively large samples are required to detect nonrandom association, unless gene frequencies are intermediate and disequilibrium is relatively intense. This might be one reason why cases of linkage disequilibrium have so far proved to be the exception, rather than the rule, in population studies.     

Non-random associations between Lap and Pept-1 loci and gene arrangements of chromosome O in D. subobscura — experiments in laboratory populations

In this paper the well-known non-random associations between Lap and Pept-1 loci and gene arrangements of chromosome O are studied in laboratory populations of D. subobscura. An increase of the frequency of the allele Lap ^1.00, towards an equilibrium point (0.70), was found to be associated with an increase of the gene arrangement O₃₊₄. This is an accordance to the associations found in natural populations. On the contrary no such an increase was observed in populations polymorphic for Lap and Pep-1 loci but homokaryotypic for gene arrangement O₃₊₄₊₈ differing in the initial allele frequencies at these loci. Although epistatic selection cannot be completely ruled out, our results are better explained under the assumption of neutrality.     

Gypsy homologous sequences inDrosophila subobscura (gypsyDS)

Summary Characterization of sequences homologous to theDrosophila melanogaster gypsy transposable element was carried out inDrosophila subobscura (gypsyDS). They were found to be widely distributed among natural populations of this species. From Southern blot and in situ analyses, these sequences appear to be mobile in this species.GypsyDS sequences are located in both euchromatic and heterochromatic regions. A completegypsyDS sequence was isolated from aD. subobscura genomic library, and a 1.3-kb fragment which aligns with the ORF2 of theD. melanogaster gypsy element was sequenced. Comparisons of this sequence in three species (D. subobscura, D. melanogaster, and D. virilis) indicate that there is greater similarity between theD. subobscura-D. virilis sequences than betweenD. subobscura andD. melanogaster. Molecular divergence ofgypsy sequences betweenD. virilis andD. subobscura is estimated at 16 MY, whereas the most likely divergence time of these two species is more than 60 MY. These data strongly suggest thatgypsy sequences have been horizontally transferred between these species.Offprint requests to: T.M. Alberola     

ASSOCIATIONS BETWEEN MITOCHONDRIAL AND NUCLEAR GENOTYPES IN CUTTHROAT TROUT HYBRID SWARMS

We examined mtDNA and nuclear allozyme genotypes in hybrid populations formed from interbreeding of westslope cutthroat trout (Oncorhynchus clarki lewisi) and Yellowstone cutthroat trout (O. c. bouvieri). These subspecies show substantial genetic divergence (Nei's D = 0.30; mtDNA P = 0.02). Diagnostic alleles at multiple nuclear loci and two distinct mtDNA haplotypes segregate in the hybrids. Nuclear and mtDNA genotypes are largely randomly associated, although there is slight disequilibrium in both nuclear and cytonuclear measures in some samples. Consistent positive gametic disequilibria for three pairs of nuclear loci confirm one previously reported linkage, and indicate two more. Allele frequencies provide no evidence for selection on individual chromosome segments. However, westslope mtDNA haplotype frequencies exceed westslope nuclear allele frequencies in all samples. This may be explained by differences in the frequency of occurrence of reciprocal F₁ matings, by viability, fertility, or sex ratio differences in the progeny of reciprocal matings, or by weak selection on mtDNA haplotypes.     

Genetic diversity of Cryptomeria japonica using co-dominant DNA markers based on sequenced-tagged sites

 We have investigated the genetic diversity of 11 natural populations of C. japonica using 13 polymorphic STS markers. The average unbiased heterozygosities (H _e ), the average number of alleles per locus (N _a ) and the proportion of polymorphic loci (Pl) were 0.281, 1.93 and 76.92%, respectively. Coefficients of linkage disequilibrium were calculated, and no significant deviation was found except in four combinations – which might have occurred by chance alone. The fixation index (F _IS ) for 3 loci showed statistically significant values at the 1% level. The genetic differentiation between populations was only 0.047, and there were no clear geographical tendencies in the allele frequencies or the heterozygosities among populations. Consequently, the results from STS-based co-dominant DNA marker analysis were very similar to those from a previous allozyme study. However, the resolution of the technique is greater than allozyme analysis because many loci with high heterozygosities can be evaluated, and it is very simple. Therefore, the STS-based marker approach is very useful and convenient for population genetics and genome mapping of C. japonica. Received: 18 July 1998 / Accepted: 13 August 1998     

Effect of thermal environment on life histories of free living Drosophila melanogaster and D. subobscura

The variability of morphological and lifehistory traits in Drosophila melanogaster and D. subobscura, two sympatric Drosophila species of different climatic origin, were investigated with regard to seasonal and daily temperature and humidity fluctuations to assess thermal sensitivity and the responses of freeliving organisms to oscillating temperatures. Temperature and humidity were measured continuously at the site where Drosophila were observed throughout the day, and thus represent a realistic picture of the temperature environment of these animals. A phenotypic model (gaussian curve) of temperature performance was fitted to daily fecundities and provided estimates of the position of temperature optimum and of the maximum and the breadth of performance for both species. D. melanogaster had a higher optimum temperature than D. subobscura. A trade-off between the maximum and the breadth of performance was detected both within and between the species.     

Nucleotide diversity of a ND5 fragment confirms that population expansion is the most suitable explanation for the mtDNA haplotype polymorphism of <Emphasis Type="Italic">Drosophila subobscura</Emphasis>

Results from mitochondria (mt) DNA restriction site analyses (RSAs) have revealed that wild populations of Drosophila subobscura are formed by two common (I and II) and some rare, often endemic, low-frequency haplotypes. In the study reported here, we analysed nucleotide diversity in a 942-bp fragment of the mtDNA ND5 gene in 48 D. subobscura individuals captured from three populations that showed haplotypes I, II or the less common ones, as well as in one additional individual belonging to D. guanche that was taken as an outgroup. RSAs and sequencing results were compared. The two approaches yielded similar nucleotide variability parameters, suggesting a consistency in the results obtained from mtDNA dynamics in natural populations of D. subobscura. Patterns of polymorphism at ND5 are most consistent with the hypothesis of population expansion after a bottleneck that may have occurred since the last glaciation or which may occur seasonally after the summer and winter. However, we cannot rule out that selection has a role in maintaining the two major haplotypes at intermediate frequencies in worldwide populations of D. subobscura.     

Genetic Coadaptation in the Chromosomal Polymorphism of DROSOPHILA SUBOBSCURA. I. Seasonal Changes of Gametic Disequilibrium in a Natural Population

Seasonal changes in gene arrangement and allozyme frequencies have been investigated in Drosophila subobscura for several years. Some arrangements (O_st and O₃₊₄₊₇) show seasonal variation, which suggests that chromosomal polymorphism is flexible in this species. Seasonal changes in allozyme frequencies for Lap and Pept-1 loci, both located within the same inversions of chromosome O, are significant only inside the O_st arrangement, but not inside O₃₊₄ arrangement. This arrangement-dependent response of allozyme generates variation in arrangement-allozyme disequilibrium. The historical hypothesis on the maintenance of disequilibria cannot explain these seasonal changes, and some kind of natural selection must be invoked. Association between Lap and Pept-1 is also seasonal inside O_st but not inside O₃₊₄. We propose that O_st probably consists of a finite array of supergenes that are differentially favored in each season by natural selection. The present evidence on this supergene selection and other genetic, biogeographic and phylogenetic data points to O₃₊₄ as the most primitive gene order among the present arrangements.     

Studies of the species barrier between Drosophila subobscura and D. madeirensis IV. A genetic dissection of the X chromosome for speciation genes

The genetics of four traits contributing to the isolation mechanism between the two closely related species of Drosophila belonging to the obscura group, D. subobscura and D. madeirensis, have been investigated, especially regarding the influence exerted by the X chromosome. This chromosome has been roughly dissected genetically by the use of four markers. It was found that factors affecting viability of backcross males are spread from the centromeric end of the chromosome up the region marked by Bx. Three sections were responsible for male sterility/fertility. The abnormal head shape of the backcross males was affected by factor(s) on the madeirensis and the subobscura sex chromosome located at the region of MAD1 inversion. Finally, an abnormal trait in these males (presence of extra sex combs) was found to be controlled by four sections, two on the madeirensis X chromosome and two on the subobscura one.