Gametic associations between inversion and allozyme polymorphisms in Drosophila buzzatii.

Gametic disequilibria between second chromosome polymorphic arrangements and seven linked allozyme loci were estimated in seven populations of Drosophila buzzatii from Argentina. Significant and consistent associations across populations were detected for Est-1, Est-2, Aldox, and XDH: Phenograms based on Nei's genetic distance showed that chromosomes carrying the 2ST arrangement were more similar to each other, irrespective of the population from which they were extracted, than to chromosomes carrying the derived 2J and 2JZ3.Restriction of recombination in heterokaryotypes seems to be the best explanation for the significant linkage disequilibria between inversions and the loci located inside the rearranged segments, for example, Est-1 and Aldox, or close to the break points, for example, Est-2. However, epistatic interactions between Xdh, which is outside the inversions and not near the break points, and loci tightly linked to the inversions, is the most likely explanation for the association between Xdh and chromosomal arrangements. Some of the associations detected in endemic Argentinean populations are coincident with data obtained in colonizing populations of the Old World and Australia. Thus historical processes that took place in the original area of the species' distribution can account for these linkage disequilibria in colonized populations of D. buzzatii.     

A study of wing morphology and fluctuating asymmetry in interspecific hybrids between Drosophila buzzatii and D. koepferae

In this work we investigate the effect of interspecific hybridization on wing morphology using geometric morphometrics in the cactophilic sibling species D. buzzatii and D. koepferae. Wing morphology in F₁ hybrids exhibited an important degree of phenotypic plasticity and differs significantly from both parental species. However, the pattern of morphological variation between hybrids and the parental strains varied between wing size and wing shape, across rearing media, sexes, and crosses, suggesting a complex genetic architecture underlying divergence in wing morphology. Even though there was significant fluctuating asymmetry for both, wing size and shape in F₁ hybrids and both parental species, there was no evidence of an increased degree of fluctuating asymmetry in hybrids as compared to parental species. These results are interpreted in terms of developmental stability as a function of a balance between levels of heterozygosity and the disruption of coadaptation as an indirect consequence of genomic divergence.     

A three-locus system of interspecific incompatibility underlies male inviability in hybrids between Drosophila buzzatii and D. koepferae

In hybrids between the sibling species D. buzzatii and D. koepferae, both sexes are more or less equally viable in the F₁: However, backcross males to D. buzzatii are frequently inviable, apparently because of interspecific genetic incompatibilities that are cryptic in the F₁. We have performed a genetic dissection of the effects of the X chromosome from D. koepferae. We found only two cytological regions, termed hmi-1 and hmi-2, altogether representing 9% of the whole chromosome, which when introgressed into D. buzzatii cause inviability of hybrid males. Observation of the pattern of asynapsis of polytene chromosomes (incomplete pairing, marking introgressed material) in females and segregation analyses were the technique used to infer the X chromosome regions responsible for this hybrid male inviability. The comparison of these results with those previously obtained with the same technique for hybrid male sterility in this same species pair indicate that in the X chromosome of D. koepferae there are at least seven times more regions that produce hybrid male sterility than hybrid male inviability. We have also found that the inviability brought about by the introgression of hmi-1 is suppressed by the cointrogression of two autosomal sections from D. koepferae. Apparently, these three regions conform to a system of species-specific complementary factors involved in an X-autosome interaction that, when disrupted in backcross hybrids by recombination with the genome of its sibling D. buzzatii, brings about hybrid male inviability.     

Larval age differences and competition between Drosophila aldrichi and D.buzzatii

Abstract.

• 1 Competitive interactions among larvae of Drosophila species using the same habitat will usually involve individuals varying in age and size. Older, larger larvae might be expected to outcompete younger, smaller larvae.
• 2 Newly hatched laryae of Drosophila aldrichi and D.buzzatii were placed on induced cactus necroses (simulating the natural habitat), either simultaneously, or with a 1 or 2 day difference in the time at which individuals of the other species were added.
• 3 Evidence for competitive differences with larval age were found. In both species, body weight was significantly higher for those larvae added first than for later addition larvae, and a longer time to complete development was required for those larvae added 2 days later. However, pre-adult viability of D.aldrichi was significantly higher for larvae started 1 or 2 days after larvae of D.buzzatii. For D.buzzatii, viability was lowest for larvae started simultaneously with D.aldrichi, but not different between those started either before or after D.aldrichi.
• 4 Considering all three traits, no clear advantage for the early addition larvae was observed.

     

Transpecific polymorphisms in an inversion linked esterase locus in Drosophila buzzatii

Nucleotide variation was studied in a 1.1 kb section of the coding region of an Esterase gene (Est-A) that maps in the center of the segments rearranged by polymorphic inversions in the cactophilic Drosophila buzzatii. We examine 30 homozygous second-chromosome lines differing in gene arrangement and three D. koepferae isofemale lines as outgroups. Our data show that Est-A is a highly polymorphic gene at both synonymous and replacement sites. Significant departures from homogeneity in the distribution of the ratio of silent polymorphism to divergence predicted by the neutral theory reveals a local excess of silent polymorphism. This is consistent with the presence of two apparent narrow peaks of elevated silent polymorphism surrounding nonconservative amino acid substitutions. These polymorphisms as well as others at synonymous and nonsynonymous sites are shared with D. koepferae. We suggest that the presence of shared nucleotide polymorphisms is probably due to interspecific gene flow and/or balancing selection acting on replacement variants and/or to a decreased probability of loss of ancestral polymorphisms caused by linkage to an adaptive inversion polymorphism. Recurrent mutation and persistence of neutral ancestral polymorphisms cannot, however, be ruled out. The analysis of the distribution of nucleotide variation among the three chromosomal arrangements sampled reveals that derived arrangements (J and JZ(3)) are less polymorphic than the ancestral ST, and that the widely distributed ST and J arrangements are genetically differentiated. However, a significant number of polymorphisms are shared between arrangements, suggesting frequent exchange either from gene conversion or from double crossovers in heterokaryotypes. Finally, our present results in combination with data of sequence variation at the breakpoints of inversion J suggest that this old gene arrangement has risen in frequency in relatively recent times.     

Divergence at neutral and non-neutral loci in Drosophila buzzatii populations and their hybrids

The impact of intraspecific hybridisation on fitness and morphological traits depends on the history of natural selection and genetic drift, which may have led to differently coadapted gene-complexes in the parental populations. The divergence at neutral and non-neutral loci between populations can be evaluated by estimating F_ST and Q_ST respectively, and hence give an estimate of drift and selection in the populations. Here we investigate (1) whether divergence between populations in quantitative traits (wing size and shape) can be attributed to selection or drift alone, (2) The impact of intraspecific hybridisation on estimators for divergence at neutral (F_ST) and non-neutral loci (Q_ST) in hybrids, (3) If measurement of shape is more informative than size in order to detect divergence in quantitative traits between populations. The aims were addressed by performing two hybridisations between three populations of Drosophila buzzatii, one between populations from Argentina and the Canary Islands (separated for 200 years), and the other between populations from Argentina and Australia (separated for 80 years). We observed the highest divergence at neutral loci between the Argentinean and Canary Island populations, but highest morphological divergence between the Argentinean and Australian populations, indicating that natural selection is acting on the wings. Divergence based on Q_ST measures in the hybrids was sensitive towards increased phenotypic variance (σ²p) within groups and should be used with care when σ²p of populations differ. Our results indicate that measures of shape give a better estimate of divergence at the underlying quantitative traits loci than measures of size.     

The evolutionary history of Drosophila buzzatii. XXIII. High content of nonsatellite repetitive DNA in D. buzzatii and in its sibling D. koepferae.

The frequency and types of repetitive nonsatellite DNA of two sibling species of the repleta group of Drosophila, D. buzzatii, and D. koepferae have been determined. For each species, the analysis is based on a sample of more than 100 clones (400 kb) obtained from genomic DNA. A theoretical model has been developed to correct for the presence of a mixture of repetitive and unique DNA in these clones. After correction, a high content of repetitive DNA has been demonstrated for both species (D. buzzatii, 19-26%; D. koepferae, 27-32%). The repetitive sequences have been classified according to their hybridization pattern when used as probes against genomic DNA and by their in situ hybridization signals on polytene chromosomes. Data suggest that the main nonsatellite component of these species is simpler and more repetitive than that of D. melanogaster, pointing to a wide variability in content and class size distribution of repetitive DNA among Drosophila species.     

Temporal and spatial variation of inversion polymorphism in two natural populations of Drosophila buzzatii

The inversion polymorphism of the cactophilic fly Drosophila buzzatii was studied in two natural populations. We assessed the temporal changes and microspatial population structure. We observed a significant increase in the frequency of arrangement 2J at the expense of 2ST in both populations. These gene arrangements appear to affect the life-history of flies differently. Environmental heterogeneity explains the karyotype coexistence in nature. The analysis of population structure showed that differentiation of inversion frequencies among individual breeding sites, the rotting clacodes of Opuntia vulgaris, was highly significant. The karyotypic frequencies did not depart significantly from Hardy-Weinberg expectations, neither in individual rots nor in the total population. These results suggest that the observed population structure can be easily accounted by random genetic drift.     

Contrasting population genetic structures using allozymes and the inversion polymorphism in Drosophila buzzatii

Second chromosome inversion and genotypic frequencies at seven allozyme loci, differentially associated with inversions, were determined in seven natural populations of Drosophila buzzatii. The patterns of variation of allozymes and the inversion polymorphisms were significantly different, indicating the role of adaptive differentiation for the latter. Moreover, the patterns of population structure varied among allozyme loci, suggesting the operation of diversifying selection for certain loci. Differentiation was negligible for Leucyl‐amino peptidase (Lap) and Peptidase‐2 (Pep‐2), low to moderate for Aldehyde oxidase (Aldox), Peptidase‐1 (Pep‐1) and Esterase‐1 (Est‐1) and high for Esterase‐2 (Est‐2) and Xanthine dehydrogenase (Xdh). Significant linkage disequilibria were detected between inversions and Aldox, Est‐1, Est‐2 and Xdh. Multiple regression analyses of inversion and allele frequencies on environmental variables revealed the existence of clines for inversions, Est‐1, Est‐2, Xdh and Aldox along altitudinal, latitudinal and/or climatic gradients. Tests using conditional allele frequencies showed that Est‐1 and Aldox clines could be accounted for by hitchhiking with inversions, whereas natural selection should be invoked to explain the clines observed for Est‐2 and Xdh.     

Polymorphic inversion and esterase loci complex on chromosome 2 of Drosophila buzzatii. II. Spatial variation

The potential influence of linked inversions on allele frequency variation at the Est-1 and Est-2 loci among Australian populations of D. buzzatii was determined by statistical analyses allele and inversion gametic frequencies. Most of the significant spatial and climatic associations found for all Est-1 allele frequencies, and for one allele only of Est-2 (Est-2c+), were accounted for by their linkage disequilibria with the inversions, which covaried with environmental variables. Consistent with this result, the spatial and climatic associations for conditional Est-1 and Est-2 allele frequencies tended to be different from those for the respective unadjusted allele frequencies. In one important respect, the results for Est-1 and Est-2 were not altered by inversions. For both unadjusted and conditional Est-1 allele frequencies, few climatic associations remain after correcting for geographic location, whereas for both unadjusted and conditional Est-2 allele frequencies, climatic associations remain after correcting for geographic location. Thus, apparent selection affecting allele frequencies at the Est-2 locus is not accounted for by linked inversions.     

A system of complementary genes in hybrids between Drosophila koepferae and D. buzzatii: a Markov chain model allows to make inferences about their number and relationships

In backcrosses between D. koepferae and D. buzzatii, the disruption of a system of species-specific complementary factors brings about hybrid male inviability. This system consists of a lethal factor, hmi-1, linked to the X chromosome of D. koepferae, and several conspecific autosomal suppressors. However, hmi-1 hybrid males can also be rescued by factors present in some strains of D. buzzatii. The present work aims to estimate the number of hmi-1 suppressors in one of these strains by means of Markov chains. The obtained results allow discarding models with one or more chromosomes having independent suppressor effect. On the other hand, models having n chromosomes that interact in groups of r, being 1 < r < or = n, to produce rescue effect, provide good approximations to the observed results. The best fit to the data is obtained with four or five chromosomes with suppressor effect, interacting epistatically in groups of three to rescue the viability of hmi-1 males.     

Nested cladistic analysis of Brazilian populations of Drosophila serido.

The effects of Quaternary climatic cycles were investigated in Drosophila serido, a Brazilian cactophilic fly widely distributed outside the Amazonian region. Previous studies have indicated this species displays remarkable karyotypic, male genitalia, and mtDNA variation, so much so that it has been described as a species complex, or superspecies. In the present study we expand the analysis of the mtDNA COI gene on D. serido populations, particularly in central Brazil, by obtaining DNA sequences from 248 individuals distributed across 47 localities. This allowed us to perform a nested clade analysis to discriminate historical from recurrent forces shaping the evolution of D. serido populations. The nested analysis indicates one event of past fragmentation separating populations from south and central Brazil (referred to as type B) from populations in central and northeast Brazil (type D) and 15 other significant events. The most common outcome of our analysis was contiguous range expansion and we discuss why this was expected in D. serido. Our data indicate that D. serido has been distributed across Brazil at least since the Mid-Pleistocene, which contradicts the hypothesis of current distribution being determined by last glaciation cycle. Nonetheless, we present evidence that climatic cycles during the Quaternary and before have had a significant impact on the differentiation of D. serido in Brazil. Our study confirms the usefulness of the nested clade analysis for disentangling the effects of historical and present-day forces shaping the evolution and distribution of a taxon.     

Genetic analysis of extra sex combs in the hybrids between Drosophila subobscura and D. madeirensis

Drosophila subobscura and D. madeirensis are closely related species, the first distributed over a large area and the latter restricted to the island of Madeira. These species can hybridize in laboratory conditions, yielding fertile females and sterile males. Hybrid offspring show several phenotypic anomalies, including sex combs on the second and third pairs of legs in males. The extra sex comb trait is a homeotic transformation of the mesothoracic and metathoracic legs into prothoracic legs. This anomaly is observed almost exclusively in F₁ males with D. madeirensis mothers. Analysis of backcross males shows that D. subobscura and D. madeirensis have diverged at a minimum of four autosomal loci affecting the extra sex comb anomaly. In addition, some incompatibility involving the X chromosome and/or a maternal effect is also implicated.     

Genetic variation and plasticity of thorax length and wing length in Drosophila aldrichi and D. buzzatii

Reaction norms across three temperatures of development were measured for thorax length, wing length and wing length/thorax length ratio for ten isofemale lines from each of two populations of Drosophila aldrichi and D. buzzatii. Means for thorax and wing length in both species were larger at 24 °C than at either 18 °C or 31 °C, with the reduction in size at 18 °C most likely due to a nutritional constraint. Although females were larger than males, the sexes were not different for wing length/thorax length ratio. The plasticity of the traits differed between species and between populations of each species, with genetic variation in plasticity similar for the two species from one locality, but much higher for D. aldrichi from the other. Estimates of heritabilities for D. aldrichi generally were higher at 18 °C and 24 °C than at 31 °C, but for D. buzzatii they were highest at 31 °C, although heritabilities were not significantly different between species at any temperature. Additive genetic variances for D. aldrichi showed trends similar to that for heritability, being highest at 18 °C and decreasing as temperature increased. For D. buzzatii, however, additive genetic variances were lowest at 24 °C. These results are suggestive that genetic variation for body size characters is increased in more stressful environments. Thorax and wing lengths showed significant genetic correlations that were not different between the species, but the genetic correlations between each of these traits and their ratio were significantly different. For D. aldrichi, genetic variation in the wing length/thorax length ratio was due primarily to variation in thorax length, while for D. buzzatii, it was due primarily to variation in wing length. The wing length/thorax length ratio, which is the inverse of wing loading, decreased linearly as temperature increased, and it is suggested that this ratio may be of greater adaptive significance than either of its components.     

Patterns of variation in wing morphology in the cactophilic Drosophila buzzatii and its sibling D. koepferae

Drosophila buzzatii and D. koepferae are two sibling species that breed on the necrotic tissues of several cactus species and show a certain degree of niche overlap. Also, they show differences in several life history traits, such as body size and developmental time, which probably evolved as a consequence of adaptation to different host plants. In this work we investigate the ecological and genetic factors affecting wing morphology variation both within and between species. Three wing traits were scored, distal and proximal wing length and width in isofemale lines reared in two of the most important host cacti: Opuntia sulphurea and Trichocereus terschekii. Our results revealed that differences between species and sexes in wing size and shape were significant, whereas the cactus factor was only significant for wing size. Intraspecific analyses showed that differences among isofemale lines were highly significant for both size and shape in both species, suggesting that an important fraction of variation in wing morphology has a genetic basis. Moreover, the line by cactus interaction, which can be interpreted as a genotype by environment interaction, also accounted for a significant proportion of variation. In summary, our study shows that wing size is phenotypically plastic and that populations of D. buzzatii and D. koepferae harbour substantial amounts of genetic variation for wing size and shape. Interspecific differences in wing size and shape are interpreted in terms of spatial predictability of the different host plants in nature.     

Embryonic lethality leads to hybrid male inviability in hybrids between Drosophila melanogaster and D. santomea

The study of the morphological defects unique to interspecific hybrids can reveal which developmental pathways have diverged between species. Drosophila melanogaster and D. santomea diverged more than 10 million years ago, and when crossed produce sterile adult females. Adult hybrid males are absent from all interspecific crosses. We aimed to determine the fate of these hybrid males. To do so, we tracked the development of hybrid females and males using classic genetic markers and techniques. We found that hybrid males die predominantly as embryos with severe segment‐specification defects while a large proportion of hybrid females embryos hatch and survive to adulthood. In particular, we show that most male embryos show a characteristic abdominal ablation phenotype, not observed in either parental species. This suggests that sex‐specific embryonic developmental defects eliminate hybrid males in this interspecific cross. The study of the developmental abnormalities that occur in hybrids can lead to the understanding of cryptic molecular divergence between species sharing a conserved body plan.     

Synapsis and recombination in inversion heterozygotes

Inversion heterozygotes are expected to suffer from reduced fertility and a high incidence of chromosomally unbalanced gametes due to recombination within the inverted region. Non-homologous synapsis of the inverted regions can prevent recombination there and diminish the deleterious effects of inversion heterozygosity. The choice between non-homologous and homologous synapsis depends on the size of inversion, its genetic content, its location in relation to the centromere and telomere, and genetic background. In addition, there is a class of inversions in which homologous synapsis is gradually replaced by non-homologous synapsis during meiotic progression. This process is called synaptic adjustment. The degree of synaptic adjustment depends critically on the presence and location of the COs (crossovers) within the inversion loop. Only bivalents without COs within the loop and those with COs in the middle of the inversion can be completely adjusted and became linear.     

Clonal analysis in hybrids between Drosophila melanogaster and Drosophila simulans

We have analysed the viability of cellular clones induced by mitotic recombination in Drosophila melanogaster/D. simulans hybrid females during larval growth. These clones contain a portion of either melanogaster or simulans genomes in homozygosity. Analysis has been carried out for the X and the second chromosomes, as well as for the 3L chromosome arm. Clones were not found in certain structures, and in others they appeared in a very low frequency. Only in abdominal tergites was a significant number of clones observed, although their frequency was lower than in melanogaster abdomens. The bigger the portion of the genome that is homozygous, the less viable is the recombinant melano-gaster/simulans hybrid clone. The few clones that appeared may represent cases in which mitotic recombination took place in distal chromosome intervals, so that the clones contained a small portion of either melanogaster or simulans chromosomes in homozygosity. Moreover, Lhr, a gene of D. simulans that suppresses the lethality of male and female melanogaster/simulans hybrids, does not suppress the lethality of the recombinant melanogaster/simulans clones. Thus, it appears that there is not just a single gene, but at least one per tested chromosome arm (and maybe more) that cause hybrid lethality. Therefore, the two species, D. melanogaster and D. simulans, have diverged to such a degree that the absence of part of the genome of one species cannot be substituted by the corresponding part of the genome of the other, probably due to the formation of co-adapted gene complexes in both species following their divergent evolution after speciation. The disruption of those coadapted gene complexes would cause the lethality of the recombinant hybrid clones.