Patterns of puffing activity and chromosomal polymorphism in Drosophila subobscura

Summary The effect of inbreeding on the puffing patterns of polytene chromosomes of Drosophila subobscura was analysed. Puffing activity was studied in two strains of D. subobscura: one which had been subject to inbreeding for 288 generations, and in the hybrids from a cross between them. A strong overall decrease in puffing activity was found in the inbred line. In general, hybrids behaved in a similar way to the inbred line or showed activity intermediate between the two lines. The fertility and viability of the two homozygous lines and of the hybrids were also determined. These parameters of fitness are very low in the inbred line. Hybrids displayed intermediate behaviour.Publication No. 109, Departamento de Genética, Universidad de Valencia, Spain     

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.     

Patterns of puffing activity and chromosomal polymorphism in Drosophila subobscura II. Puffing patterns at the prepupa stage

Puffing activity patterns of the five large polytene chromosomes of Drosophila subobscura were studied during the late third-larval instar and through the prepupal period. A total of 166 loci active in some of the eleven stages studied were described. The distribution of these active loci per chromosome is the following: 25 on chromosome A, 33 on chromosome J, 31 on chromosome U, 34 on chromosome E and 43 on chromosome O. Seven principal patterns of puffing activity were defined taking into account the different curves of the puffing histograms. Gene activities per chromosome as well as total were analysed. Three peaks of gene activity at the beginning, middle and ending of prepupation can be observed. U is the most active chromosome and A (the sex chromosome), and J the least active. Chromosomes E and O show a medium activity. A possible biological explanation for these results is discussed.Publication No. 107, Departamento de Genética, Universidad de Valencia (Spain).     

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.     

Patterns of puffing activity in the salivary gland chromosomes of Drosophila

Exposure of Drosophila melanogaster larvae to high temperature for short periods of time results in marked changes in the puffing patterns of salivary gland chromosomes. Temperature shock induces puffing at 9 specific loci; this pattern of induced puffs shows little developmental specificity and is similar in three strains of D. melanogaster (including the mutant lethal giant-larvae) and in D. simulans. Temperature shock also (i) retards the regression of some developmentally specific puffs and (ii) results in the regression of all other puffs normal to development. The effect of temperature treatment is similar in vivo and after in vitro treatment of salivary glands. The in vitro response is not sensitive to cycloheximide. A similar pattern of induced puffs to that found after temperature treatment is found during recovery of larvae from anoxia, but additional puffs are induced after anoxia. The size and duration of activity of the induced puffs is dependent upon the magnitude of the treatment.     

Patterns of puffing activity in the salivary gland chromosomes of Drosophila

Salivary gland X chromosome puffing patterns are described for the Oregon stock of Drosophila melanogaster and for the Berkeley stock of D. simulans. In D. melanogaster regular phase specific puffing was recorded at 21 loci in the third larval instar and subsequent prepupal stage. A comparison of the X chromosome puffing patterns of male and female larvae failed to show any qualitative differences although in the males a group of puffs were active for a longer time during development than in females. The X chromosome puffing patterns of D. simulans are similar to those described for D. melanogaster although two puffs (4F 1–4 and 7B 1–3) were active in D. simulans but not in D. melanogaster. The sex differences in puffing observed in D. melanogaster were also observed in D. simulans.     

Patterns of puffing activity in the salivary gland chromosomes of Drosophila

The patterns of puffing activity in the proximal region of 2L of D. melanogaster have been reinvestigated and revised. Possible relationships between three puffs and the structural genes for alcohol dehydrogenase, dopa decarboxylase and the histones are discussed.     

Patterns of puffing activity in the salivary gland chromosomes of Drosophila

The autosomal salivary gland chromosome puffing patterns of Drosophila simulans are described and compared with the puffing patterns of the sibling species D. melanogaster. During the late third larval instar and the prepupal period the patterns of puffing activity of these two species are similar — approximately 50% of the puffs common to both species showing identical activities. The remaining puffs differ in their timing of activity, or in their mean sizes, or in both of these parameters. A number of puffs (14) found in D. simulans have not been regularly observed in the Oregon stock of D. melanogaster but are active in other D. melanogaster strains. One puff (46 A) of D. melanogaster was absent from D. simulans and forms a heterozygous puff in hybrids, when the homologous chromosomes are synapsed. When the homologues are asynapsed a puff at 46 A is restricted to the melanogaster homologue. The puff at 63E on chromosome arm 3L is considerably smaller in D. simulans than in D. melanogaster and this size difference is autonomous in hybrids. Other puffs not common to both species behave non-autonomously in the species hybrid, even when the homologous chromosomes are asynapsed.     

Patterns of puffing activity in the salivary gland chromosomes of Drosophila

A technique for the short term organ culture of larval salivary glands of D. melanogaster is described. Cultured Puff Stage 1 glands respond to 20-OH ecdysone by initiating the cycle of puffing activity characteristic of late larval development and puparium formation. This puffing cycle involves the sequential activation of at least 125 puffs. Their response to ecdysone allows these puffs to be divided into 3 main classes: a) PS1 puffs that regress (e.g. 25AC); b) puffs activated very rapidly (within 5 min) (e.g. 23E, 74EF, 75B) and c) puffs activated only after longer periods (>4 h) (e.g. 62E, 78D, 22C, 63E and 82F). The detailed behaviour of representatives of each class is described. These data support Clever's distinction of ‘early’ and ‘late’ ecdysone responsive sites.     

Patterns of puffing activity in the salivary gland chromosomes of Drosophila

Patterns of puffing activity during the third larval instar and the prepupal period of two different strains of D. melanogaster (Oregon and vg6) are compared. The variation in puffing activity observed is both quantitative (involving the mean size or timing of activity of individual puffs) and qualitative. The pattern of activity of 64% of the puffs is the same in the two strains, 12% show strain differences in puff size and 19% in the time of their activity. One puff (64C) is active only in one of the strains (vg6). In genetic experiments this puff segregates normally and the puff locus has been mapped genetically to a site coincident with, or at least very close to, the cytogenetic position of the puff. In heterozygotes the puff is homozygous only when the maternal and paternal homologues are synapsed. When the homologues are asynapsed only the homologue from the vg6 parent is puffed at 64C. With the exeption of some strains closely related to vg6 no other strain of D. melanogaster has been found to possess puffing activity at 64C. In vg6/In(3LR)C165 heterozygotes 64C forms a heterozygous puff even when the homologues are synapsed. In the discussion consideration is given to the various factors that control puff size.     

Patterns of puffing activity in the salivary gland chromosomes of Drosophila

The patterns of puffing activity have been studied during the late larval and prepupal stages of Drosophila melanogaster. On the major salivary gland autosomes (chromosomes 2 and 3) 108 loci form puffs at some time during these developmental stages. The timing and pattern of activity of 83 of these puffs is found to be strictly dependent upon the age of the animals. Two major peaks in puffing activity occur. The first of these is at the time of puparium formation and the second in 8 hr. old prepupae. Both of these puffing peaks precede a moult by 4 hrs. 30 puffs are active before or at the time of both of these two moults. However, the sequence of appearance and regression of many of this group of puffs is different at the prepupal moult than at the pupal moult. 12 puffs occur only before or at the time of the prepupal moult and 13 puffs only before or at the time of the pupal moult. The functional significance of these periods of puffing activity is discussed and it is concluded that one function of this genetic activity in the salivary glands of metamorphosing Drosophila is the production of substances to be utilised during the histogenesis of the adult tissues.     

Patterns of puffing activity in the salivary gland chromosomes of Drosophila

Puffing patterns of chromosome arm 3 L of D. yakuba are compared with those of other members of the melanogaster species subgroup D. melanogaster and D. simulans. Several paracentric inversions on 3L have resulted in a considerable rearrangement of gene order in D. yakuba. However the basic sequence of changes in puffing activity which occurs during late larval and prepupal development is very similar to that of D. melanogaster and D. simulans. A fourth member of this species subgroup (D. teissieri) also has similar puffing patterns to those of D. melanogaster despite considerable chromosome evolution.     

Monitoring chromosomal polymorphism in Drosophila subobscura over 40 years

The inversion chromosomal polymorphism of Drosophila subobscura is considered to be adaptive as a result of its responses at different time scales to temperature changes. This work reports the longest‐term study of chromosomal polymorphism for a single population of D. subobscura with climatic data from the collecting site itself. The chromosomal analysis of D. subobscura samples collected six times over a 40‐year period at the same location and in the same seasonal interval has revealed the continuous presence of 16 common and six moderately rare chromosomal arrangements through the period. This analysis also corroborates the previously detected negative relationship between the frequencies of the standard (cold‐climate) arrangement on each of its five chromosomes and temperature, as well as between a comprehensive measure of cold adaptation (the total autosomal proportion of standard arrangement) and temperature. These and previous results would support that species harboring cold‐ and warm‐adapted polymorphic chromosomal arrangements, like D. subobscura, can rapidly respond to environmental changes.     

Long-term changes in the chromosomal inversion polymorphism of Drosophila subobscura. I. Mediterranean populations from southwestern Europe

The chromosomal polymorphism of seven Mediterranean populations of Drosophila subobscura has been compared with that of the same populations collected 26 to 35 years ago. Significant latitudinal clines for the frequencies of A(ST), E(ST), O(ST). and U(ST) chromosomal arrangements have been detected in the old and new samples. Standard gene arrangements are frequent in the north and decrease in frequency towards the south. Significant negative regression coefficients between latitude and transformed frequency have also been observed for the more frequent nonstandard gene arrangements. The pattern of the clines is practically the same in the old and new collections. Furthermore, the frequencies of gene arrangements of all chromosomes have changed significantly during this period in a systematic way: an increase in the frequency of those arrangements typical of southern latitudes and a decrease for those more common in northern latitudes is observed in all populations. These changes could be due to climatic factors that are correlated with latitude, making the chromosomal composition of this species more "southern.'     

Temporal changes of chromosomal polymorphism in natural populations of Drosophila subobscura

The behaviour of the chromosomal polymorphism of D. subobscura was analyzed in relation to temporal changes, daily, seasonal and annual. Firstly, chromosome analyses were carried out over a year in a natural population. Samples were taken at monthly intervals in Tibidabo, a locality close to Barcelona. In all the months except January, the number of individuals captured was enough to carry out a chromosome analysis of that population. The A, E and O chromosomes showed a great uniformity in the frequencies of gene arrangement over the year. However, significant changes occur in the frequencies of J and U chromosomes. The J₁ and U₁₊₂₊₈ arrangements showed a similar tendency, with two maxima, in June and February, and a minimum in September. These changes seem to be unrelated to the cyclical climatic changes. A chromosome analysis of the subsamples captured during the day, in the May sample, was done. In no case was the change in chromosome frequencies statistically significant. The behaviour of the U_st arrangement must be noted, the frequency of which decreased through the day. Finally, the data for the Tibidabo population were compared with samples captured in nearby localities over a period of 21 years. Significant differences were found in the frequencies of all the chromosomes, with the exception of J. The most differentiated population was the last sampled, from 1982. In this population the st arrangements tended to decrease in the A, E and O chromosomes and A₂, E_1+2+9+12 and O₃₊₄₊₇ increase, respectively.     

Relationship between chromosomal polymorphism and wing size in a natural population of Drosophila subobscura

Chromosomal polymorphism and wing size (as a measure of body size) were analysed simultaneously in two samples of Drosophila subobscura from Barcelona, Spain. The very rich chromosomal polymorphism of this species makes it difficult to detect the relationship of this polymorphism with any phenotypical character. However, a positive significant regression of wing size on the percentage of the autosome length with standard arrangement was found. Furthermore, for each polymorphic chromosome, except for the J chromosome, an association between the most frequent arrangements and wing size was observed. This trend, which was the same in the two samples, was that expected according to the latitudinal clines of both characters.     

Differential flight activity among karyotypes: daily and weather-induced changes in chromosomal inversion polymorphism in natural populations of Drosophila subobscura

The question is posed whether karyotype frequencies in the active fraction of Drosophila subobscura populations vary consistently in the course of a day, or in accordance with weather changes. Significant daily variation occurred in all chromosomes with the exception of the sex chromosome. The proportions of the karyotypes U_st/1+2 and I_st/st decreased from early to late morning and increased during the evening. I_st/1 displayed a complementary behaviour. The homokaryotypes O_st/st and E_st/st decreased both during the morning and the evening. Weather disturbances caused a remarkable variation in the frequencies of the standard gene arrangements.     

Molecular and chromosomal polymorphism in continental and insular populations from the southwestern range of Drosophila subobscura

Eight insular and continental populations from the south-western range of Drosophila subobscura have been studied with regard to molecular and inversion polymorphisms. Heterogeneity between populations with respect to allele frequencies of 4 gene loci (Amy, Est-8, Est-9 and Pep-1) is the highest known for natural populations of the species. Moreover, the most common allele for these loci is not the same in all populations. Cladograms based on UPGMA clustering of the genetic distances based on allele frequency do not coincide with those constructed with inversion data. The allele-frequency differences between the populations may be due to non-random associations between enzyme alleles and gene arrangements and to founder effects appearing in the insular populations.     

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.