Chromosomal inversion polymorphism of Drosophila subobscura from south‐eastern part of Europe

Five Drosophila subobscura populations from south‐eastern Europe were investigated with respect to chromosomal inversion polymorphism. They belong to central European populations and their gene arrangement frequencies are in accordance with latitudinal variation observed in this species. The new complex gene arrangement A₂₊₈₊₉, found in one of the populations analysed, may be specific for the southern Balkan refugium. A general tendency of gene arrangement frequencies to exhibit some southern characteristics was found for some of the autosomal standard arrangements.     

Chromosomal inversion polymorphism of Drosophila subobscura populations from Jastrebac Mountain shows temporal and habitat-related changes

Inversion polymorphism in populations of D. subobscura from a beech forest on Jastrebac mountain was studied in June 1990, 1993, and 1994, respectively. The same analysis was performed in 1990 for D. subobscura populations in a beech forest and an oak forest in the same region. Statistically significant differences in the frequencies of the gene arrangements of A₁, J and U chromosome were observed during the period of investigation. A tendency towards a decrease in the frequency of the standard gene arrangements was found for all chromosomes, but was particularly evident with chromosomes A and J. The frequency of the gene arrangements A₁ A₂, J₁ and U_1–2+6 increased at the same time. Differences in the frequency of the gene arrangements of A, J and U chromosomes were also observed when the populations from two ecologically different habitats (beech and oak forest) were compared in 1990.     

Multilocus analysis of nucleotide variation in Drosophila madeirensis, an endemic species of the Laurisilva forest in Madeira

Drosophila madeirensis is an endemic species of Madeira that inhabits the island Laurisilva forest. Nucleotide variation in D. madeirensis is analysed in six genomic regions and compared to that previously reported for the same regions in Drosophila subobscura, an abundant species in the Palearctic region that is closely related to D. madeirensis. The gene regions analysed are distributed along the O₃ inversion. The O₃ arrangement is monomorphic in D. madeirensis, and it was present in ancestral populations of D. subobscura but went extinct in this species after the origin of the derived O_ST and O₃₊₄ arrangements. Levels of nucleotide polymorphism in D. madeirensis are similar to those present in the O_ST and O₃₊₄ arrangements of D. subobscura, and the frequency spectrum is skewed towards rare variants. Purifying selection against deleterious nonsynonymous mutations is less effective in D. madeirensis. Although D. madeirensis and D. subobscura coexist at present in Madeira, no clear evidence of introgression was detected in the studied regions.     

Monitoring of the genetic structure of natural populations: change of the effective population size and inversion polymorphism in <Emphasis Type="Italic">Drosophila subobscura</Emphasis>

We analyzed changes in the genetic structure and effective population size of two ecologically distinct populations of Drosophila subobscura over several years. Population sizes of D. subobscura in beech and oak wood habitats for a period of 6 years were estimated by the capture-mark-release-recapture method. Inversion polymorphism parameters were also assessed in the same populations for a period of 3 years. Significant differences in the numbers of individuals were observed between sexes. This affected the effective population sizes between particular years. The ratio of the effective size over the cenzus dropped significantly in beech wood in 2 years. Although overall heterozygosity remained unchanged during the years in both habitats, frequencies of gene arrangements on five chromosomes show variability. After the bottleneck, some complex chromosomal arrangements appeared for the first time in both populations. Standard gene arrangements of chromosome A increased in frequency over the years in each habitat, while the complex arrangements remain rather stable and specific for each population. The results obtained indicate that the population structure may significantly change if the effective size of D. subobscura population is reduced, which is mostly related to microclimatic changes in habitats. Based on the results to date, monitoring of microevolutionary changes by using D. subobscura and its relatives seems a promising way to study the effects of global climate changes.     

Patterns of puffing activity and chromosomal polymorphism in Drosophila subobscura

The puffing patterns in polytene E chromosomes of Drosophila subobscura were followed in third-instar larvae and throughout the prepupa period. Two gene arrangements, E_st and E_1·2+9–12 were studied. A majority of puffs exhibit a similar pattern, but the puffs 61AC and 67AB behave differently in the two chromosomal arrangements, both in homozygotes and in heterozygotes. These two puffs are located at the end of the E₁₂ inversion. This position effect is an interesting phenomenon that probably is not due to a heterochromatinization effect.     

MDH-polymorphism in Drosophila subobscura: I. Selection and hitch-hiking in laboratory populations

Summary The interrelation of genic polymorphism at the Malate dehydrogenase (Mdh)-locus and chromosomal polymorphism for superimposed gene arrangements was studied in 5 experimental populations of Drosophila subobscura. It could be shown that chromosomal polymorphism is maintained by balancing selection in favour of the heterozygotes. In contrast, selection at the Mdh-locus itself seems to be of minor importance. The changes of frequency observed for the Mdh-alleles are most probably due to hitch-hiking on the gene blocks enclosed by the chromosomal structures or to selection for tightly linked genes rather than to fitness differences between Mdh-genotypes. The results may be considered as a model for the situation found in natural populations of D. subobscura.     

The genetics of Drosophila subobscura populations. 3. Inversion polymorphism and climatic factors

Summary The natural population of Drosophila subobscura of Mt. Parnes has been followed for two successive years and the frequency of the gene arrangements determined. Some changes occured in the first year but do not explain the existence of the geographical clines for these frequencies. Within the second year no changes were observed. For chromosomes J and U there is evidence for a greater fitness of the heterozygotes.Experiments on temperature effect on viability of adult male flies, on their temporal sterility, on the viability of flies during all their biological cycle and experiments on the effect of dryness on the viability of adult male flies did not show an important change of gene arrangement frequencies in the resistant individuals. Nor did diapausing females have very different gene arrangements, except for chromosome O. Cage experiments with temperature as a selective agent did not elucidate the action of this factor. It is concluded that temperature or dryness do not affect very much the polymorphic system, nor explain the existence of these clines.This polymorphic system is much more stable in D. subobscura than in D. pseudoobscura, its nearctic relative. It is also richer. It is concluded that it is historically older and helps the populations to cope with many environmental changes.     

Drosophila subobscura in Egypt and its probable derivation

In April 1978 a total of 26565 drosophilid flies were collected in Egypt. Of ten species five are new records for Egypt, while two previously reported Drosophila species were not found. Seven of the species belong to the genus Drosophila and are known to be partly, or wholly, domestic.For the first time Drosophila subobscura is reported from Egypt. The species was collected at low frequency in an orchard near Alexandria, and one specimen was trapped in a city park in Cairo. Judged from the gene arrangements of 41 males studied, Egyptian Drosophila subobscura seem likely to be derived from recent immigrants from the south coast of Asia Minor.     

Nucleotide Polymorphism at the rp49 region of Drosophila subobscura: Lack of geographic subdivision within chromosomal arrangements in Europe

Restriction-site polymorphism at the rp49 gene region has been studied in 234 lines of Drosophila subobscura representing different gene arrangements for the O chromosome. The population surveyed (El Pedroso, Spain) was sampled four times in each of two consecutive years. The data indicate that the two chromosomal classes studied, O_[ST] and O_{[3 + 4]}, are genetically differentiated in El Pedroso. Comparison of the present results with those previously obtained for two other populations further supports that, for a given chromosomal class, European populations are not genetically differentiated. This lack of differentiation at the rp49 region within O_[ST] and within O_{[3 + 4]} stands in contrast to the clear latitudinal clines found in Europe for these arrangements.     

The banding pattern of polytene chromosomes of Drosophila guanche compared with that of D. subobscura

A detailed map of the salivary gland chromosomes of Drosophila guanche is presented and compared to the standard gene arrangements of D. subobscura. Generally, the polytene chromosomc banding patterns of the two species show a high degrce of homology. Only Segment I of the sex chromosome (Chromosome A) shows marked differences. The banding pattern proposed for this segment in D. guanche could have originated from a cluster of overlapping inversions including A₁ arrangement.     

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.     

Geographic pattern of allozyme and inversion polymorphism on chromosome O of Drosophila subobscura and its evolutionary origin

The chromosome O of Drosophila subobscura was studied with respect to genetic variability at three enzyme loci (Odh, Me, and Lap-4) and with respect to inversion polymorphism. Population samples were taken from seven localities along a north-south gradient from Sweden and Scotland to Tunisia.The chromosomal analysis revealed clinal frequency changes for gene arrangements from north to south. With the enzyme loci Odh and Me allele frequencies are similar throughout the distribution range. Both loci are located outside the common inversion complex O₃₊₄. On the other hand, frequency changes parallel to those of the gene arrangements were observed for the alleles of the Lap-locus. Nonrandom associations between Lap-alleles and the superimposed gene arrangements O_ST, O₃₊₄, O₃₊₄₊₈, and O₃₊₄₊₂₃ were found. These gene arrangements differ from each other with respect to allele frequencies at the Lap-locus but for a given gene arrangement the relative frequencies of Lap-alleles remain relatively constant along the north-south gradient. Thus allele frequencies at the Lap-locus can be predicted from inversion frequencies.These observations can be interpreted in such a way that the pattern of allozyme variation within gene arrangements is due to founder effects caused by the unique origin of inversions. The gene blocks in the different inversions seem to represent more or less separated gene pools. In polymorphic populations the coexistence of genetically differentiated inversions presumably gives rise to heterotic interaction.     

Chromosomal polymorphism in natural populations of Drosophila subobscura near Zürich,Switzerland: a contribution to long-term comparisons

The chromosomal inversion polymorphism in natural populations of D. subobscura was investigated near Zürich, Seitzerland, in 1986 and 1987. These data are compared with earlier data from 1963, 1964 and 1984, collected in the same region. In all five acrocentric chromosomes, significant differences in the proportions of the dominant gene arrangements occurred; in the chromosomes A, I, O and U the standard order decreased from the sixties to the eighties, whereas the arrangements I₁, O₃₊₄, O₃₊₄₊₈ and U₁₊₂ became more frequent.     

Studies of chromosome structure polymorphism in Asia Minor and Persian populations of Drosophila subobscura Coll

Summary By means of the male sample method samples of D. subobscura from the north and south coast of Asia Minor and from the south coast of the Caspian Sea were examined with regard to the gene arrangements contained in them. The samples were judged by the following three criteria: (1) the number of different gene arrangements present, (2) the frequencies in which these gene arrangements occur, and (3) the rate of zygotic combinations.The samples from Asia Minor were found to be highly polymorphic. In this respect, they are comparable with other samples from the northern border of the Mediterranean region.The Persian populations are poor as regards number of gene arrangements. Commonly, some gene arrangements are highly frequent and, accordingly, high rates of homozygous combinations dominate. Presumably, these are marginal populations.Comparisons between samples from the north and south coast of Asia Minor confirm the findings from other parts of the species' range. As a rule, the frequencies of various gene arrangements vary according to the latitude in which they occur. Since climate and vegetation are also largely correlated with latitude, it is assumed that this is due to adaptive reaction to ecological factors.Comparing populations from the Western and Eastern Mediterranean region, similar differences were recorded. Some gene arrangements seem to be confined to either the western or eastern part of the area, and other gene arrangements differ in frequency. Since the ecological conditions in these areas appear to be comparable, such differences might be due to the history of the formation and dispersal of gene arrangements.In general, the frequencies of the gene arrangements in the chromosomes A, U, E and O are most equal along the northern margin of the Mediterranean area and, correspondingly, the frequencies of the heterokaryotypes are highest. It may be assumed that many heterokaryotypes reach their highest degree of heterotic supremacy in this area, and/or that the different homokaryotypes are uniform with respect to adaptive values. The reasoning rests on the additional assumption that in this area the biological conditions are the least extreme, or, in other words, that D. subobscura is best adapted to this environment. As it is, the region appears to lie geographically near the center of the species' range. In the I-chromosome the most equal frequencies of gene arrangements along with the highest frequency of the heterokaryotypes are to be found in Central Europe.On the other hand, in various populations lying supposedly at the borders of the species' range, the frequencies of gene arrangements of at least some chromosomes are strongly unequal and, accordingly, the frequencies of the homokaryotypes are high.In populations where the polymorphism is high, the character of the polymorphism varies slightly from chromosome to chromosome. In the O-chromosome usually one gene arrangement is abundant, while a large number of others are rare; two gene arrangements tend to dominate in the I- and U-chromosome, whereas in the E-chromosome several gene arrangements are apt to share similar frequencies.     

Linkage disequilibrium in chromosome A ofDrosophila subobscura

Evidence of strong and consistent association of alleles at theHbdh and6Pgdh loci with the A₂ and A₂₊₆ gene arrangements of the sex chromosome inD. subobscura is presented. Total association between one rare allele ofG6pdh and the rare sex-ratio arrangement A_2+3+5+7 has also been found. Selective or historical factors could explain these linkage disequilibria.     

Seasonal changes in chromosomal inversion polymorphism in a <Emphasis Type="Italic">Drosophila subobscura</Emphasis> natural population from a southeastern European continental refugium of the last glaciation period

Chromosomal inversion polymorphism was studied in a natural population of Drosophila subobscura from the Djerdap gorge of the Danube River, which is one of southeastern European continental refugia during the last glaciation period. The existence of changes in chromosomal inversion polymorphism during different periods of time was tested. The observed high abundance of chromosomal arrangements and lower IFR value allowed this population to be classified with central European populations. It was characterized by the O₇ and O_{3 + 4 + 5} arrangements, not found in other populations from the region analyzed. Within the region studied, the E_{1 + 2 + 9 + 12} chromosomal arrangement was found, apart from this population from a warm habitat, only in one population from a Mediterranean refugium. Many similarities have been observed between the population from the Djerdap gorge and other populations from the region analyzed, which indicates that postglacial migration may have started from this southeastern European refugium. Seasonal changes in the frequencies of the E_st and O_{3 + 4 + 7} chromosomal arrangements, E_st and O_{3 + 4 + 7}, being more frequent in spring and in summer, respectively, were found. The changes in the J chromosome (J_st/J_st), E chromosome (E_st/E_{1 + 2 + 9} and E₈/E_{1 + 2 + 9}), and O chromosome (O_st/O_{3 + 4 + 22}; O_{3 + 4}/O_{3 + 4}; and O_{3 + 4 + 1}/O_{3 + 4 + 7}), whose karyotypic combination frequencies were associated with seasonal changes, were also found. These changes are most probably associated with seasonal climatic changes in the population analyzed, which confirms that the chromosomal inversions in D. subobscura are adaptive. The text was submitted by the author in English.     

Latitudinal differences in sex chromosome inversions,sex linked allozymes,and mitochondrial DNA variation in Drosophila subobscura

Mitochondrial DNA, sex linked allozymes, and chromosome A gene arrangement data, from eight European natural populations of Drosophila subobscura, were analyzed to determine the existence of latitudinal clines. Strong north-south correlations with latitude were found for gene arrangements and for the Hbdh and 6Pgdh allozymes. Gametic associations between the A₂ gene arrangement, the Hbdh ⁹⁶ and the 6Pgdh ⁹⁶ alleles, point out some kind of epistatic interaction. At mtDNA level, the Hae III, A variant did not show a previously found north-south clinal variation.     

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.     

Tracing the colonization of Madeira and the Canary Islands by Drosophila subobscura through the study of the rp49 gene region

Nucleotide variation at the nuclear ribosomal protein 49 (rp49) gene region has been analysed by fine restriction mapping in a sample of 47 lines from a population from Madeira. Five restriction-site (out of 37 sites scored) and 3 length polymorphisms have been detected, resulting in 14 different haplotypes. This population shows less variation than both continental and Canary Island populations. The population from Madeira shows some differentiation from mainland populations, which does not favor the idea of extensive migration between the continent and Madeira. Chromosomal and restriction-map variation of the rp49 region in D. subobscura populations, together with data on sequence comparison of this nuclear region in D. guanche and D. madeirensis clearly indicate that the Canary Islands underwent at least two colonization events from the nearby continent. Although the data for Madeira are compatible with a single colonization event by a continental sample polymorphic for gene arrangements O₃ and O_{3 + 4}, an alternative scenario with at least two colonization events seems more likely.     

3H-uridine labelling patterns and chromosomal polymorphism inDrosophila subobscura: J and U chromosomes

The³H-uridine labelling patterns in J and U polytene chromosomes ofDrosophila subobscura were determined. The analysis was carried out in two developmental stages and in two strains proceeding from the same geographical origin whose genotypes were: J_st/J_st; U₁₊₂/U₁₊₂ and J₁/J₁; U₁₊₂₊₈/U₁₊₂₊₈ respectively. It was observed that the labelling pattern coincided very approximately with the puffing pattern in the same stages and chromosomal arrangements. Comparison of the³H-Uridine incorporation patterns between chromosomal arrangements showed light quantitative differences. These results are discussed in relation to the inversion effect.