Drosophila Buzzatii Research Articles

Allozymatic divergence between border populations of two cryptic species of the Drosophila buzzatii cluster species (Diptera: Drosophilidae)

Drosophila antonietae and Drosophila gouveai are allopatric, cactophilic, cryptic and endemic of South America species, which aedeagus morphology is considered the main diagnostic character. In this work, single close populations from the edge distributions of each species, located in an “introgressive corridor”, were analyzed regarding temporal isozenzymatic genetic variability. Isocitrate dehydrogenase ( Idh) appeared as a diagnostic locus between D. antonieate and D. gouveai because each population was fixed for different alleles. Moreover, several polymorphic loci showed accentuated divergence in the allele frequency, as evidenced by Nei’s I (0.3188) and D (1.1432), and also by Reynolds’ genetic distance and identity (1.3207 and 0.7331, respectively). Our results showed that, in spite of the very similar external morphology, related evolutionary histories, close distributions, and events of introgression in the studied area, these cryptic species have high allozymatic differentiation, and this is discussed here.

Developmental acclimation affects clinal variation in stress resistance traits in Drosophila buzzatii

Patterns of clinal genetic variation in Drosophila are often characterized after rearing at constant temperatures. However, clinal patterns might change after acclimation if populations differ in their plastic response to fluctuating environments. We studied longevity, starvation and heat knock-down resistance after development at either constant or fluctuating temperatures in nine Drosophila buzzatii populations collected along an altitudinal gradient in Tenerife, Spain. Flies that developed at fluctuating temperatures had higher stress resistance despite experiencing a slightly lower average temperature than those at constant temperatures. Genetic variation along the gradient was found in both stress-resistance traits. Because Q(ST) values greatly exceeded F(ST) values, genetic drift could not explain this diversification. In general, differences among populations were larger after rearing at fluctuating temperatures, especially in heat knock-down, for which clinal patterns disappeared when flies were reared at constant temperatures. This result emphasizes the importance of determining whether populations originating from different environments differ in their plastic responses to stress.

Intra- and interspecific divergence in the nuclear sequences of the clock gene period in species of the Drosophila buzzatii cluster

We have measured nucleotide variation in the CLOCK/CYCLE heterodimer inhibition domain (CCID) of the clock X-linked gene period in seven species belonging to the Drosophila buzzatii cluster, namely D. buzzatii, Drosophila koepferae, Drosophila antonietae, Drosophila serido, Drosophila gouveai, Drosophila seriema and Drosophila borborema. We detected that the purifying selection is the main force driving the sequence evolution in period, in agreement with the important role of CCID in clock machinery. Our survey revealed that period provides valuable phylogenetic information that allowed to resolve phylogenetic relationships among D. gouveai, D. borborema and D. seriema, which composed a polytomic clade in preliminary studies. The analysis of patterns of intraspecific variation revealed two different lineages of period in D. koepferae, probably reflecting introgressive hybridization from D. buzzatii, in concordance with previous molecular data. En este trabajo hemos medido la variación nucleotídica en el dominio CCID en el gen period, localizado en el cromosoma X, asociado a los ritmos circadianos, el enjambre de siete especies que conforman denominado “cluster”Drosophila buzzatii: D. buzzatii, D. koepferae, D. antonietae, D. serido, D. gouveai, D. seriema y D. borborema. Nuestro estudio mostró que la selección purificadora es la principal fuerza que gobierna la evolución de period, en acuerdo con una región que como CCID juega un papel importante en la maquinaria de los ritmos circadianos. Hemos observado que la variación en period provee información filogenética que permitió resolver la politomía que involucra a D. gouveai, D. borborema y D. seriema. Finalmente, el análisis de la variación intraespecifíca reveló la presencia de dos linajes en D. koepferae, uno de los cuales podría ser consecuencia de hibridación introgressiva desde D. buzzatii, en acuerdo con estudios anteriores en regiones génicas localizadas en otros cromosomas. Table S1. Polymorphism statistics and minimum number of recombination events applied to period gene sequences of the species of Drosophila buzzatii cluster. Table S2. Pairwise comparison of fixed differences and shared polymorphisms among the Drosophila buzzatii cluster species, where the shared polymorphism is shown in parenthesis (above diagonal). Average intraspecific (diagonal) and interspecific (below diagonal) genetic distances values were based on the Tamura and Nei model [Tamura and Nei (1993)]. Table S3. Results of Z-test of selection based on the Nei and Gojobori method (Nei and Gojobori, 1986). Figure S1. Transition (s) and transversion (v) rates plotted against divergence among sequences of the nuclear gene period based on the Tamura and Nei model (Tamura and Nei, 1993). Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Direct and correlated responses to chill‐coma recovery selection in Drosophila buzzatii

AbstractChill‐coma recovery (CCR) is an important trait for thermal adaptation in insects. Multiple phenotypes could be affected by selection on CCR if the trait is genetically correlated with other adaptive traits. To test for heritable (co‐)variation in CCR, we examined direct and correlated responses to bi‐directional selection on CCR. Drosophila buzzatii Patterson & Wheeler (Diptera: Drosophilidae) was artificially selected for decreased and increased recovery time following exposure to 0 °C. After 18 selected generations, the selection response in CCR was significant but qualitatively asymmetric, with replicated lines for slow CCR showing the highest response. Knockdown resistance to high temperature was not affected by CCR selection. Starvation resistance in the adult fly showed no clear pattern of correlated responses to CCR selection. Selection on CCR had no impact on developmental time and body size. Chill‐coma recovery shows no apparent genetic trade‐offs with any of the multiple traits included in this study. These results are largely consistent with recent studies on clines in D. buzzatii, which showed that CCR is not across‐population correlated with other clinally varying traits of thermal adaptation. Cold adaptation may evolve toward increased cold resistance independent of upper thermal limits.

A biophysical interpretation of temperature-dependent body size in Drosophila aldrichi and D. buzzatii

The temperature–size rule, the observation that most ectotherms grow faster but reach smaller size at higher temperatures, has defied a general explanation. Here, the temperature–size rule in Drosophila aldrichi and Drosophila buzzatii is investigated, using data on development rate and adult dry weight at nine temperatures. In both species the linear regression of dry weight on temperature is negative. The data are used to infer the potential for a description of temperature dependent size by biophysical modelling. The biophysical Sharpe–Schoolfield model for biological rates and its derivative model for adult weight yield detailed patterns for the two species’ development rate, growth rate, and adult weight. These detailed patterns do not confirm the existence of a simple temperature–size rule. The species differ significantly in the values of the parameters in the Sharpe–Schoolfield model, and as a consequence in different patterns of weight over temperatures. The different parameters of the Sharpe–Schoolfield model play distinct roles in the patterns of weight over temperatures. A temperature–size rule as a negative regression of weight on temperature might statistically follow from an upper temperature boundary for growth that is lower than the upper temperature boundary for development; as such a relation between the upper temperature boundaries for growth and development would lead to a decrease of weight at high temperature.

Characterization of the shsp genes in Drosophila buzzatii and association between the frequency of Valine mutations in hsp23 and climatic variables along a longitudinal gradient in Australia

The small heat shock gene (shsp) cluster of Drosophila buzzatii was sequenced and the gene order and DNA sequence were compared with those of the shsps in Drosophila melanogaster. The D. buzzatii shsp cluster contains an inversion and a duplication of hsp26. A phylogenetic tree was constructed based on hsp26 genes from several Drosophila species of the Sophophora and Drosophila subgenera. The tree shows first a separation of the Sophophora and the Drosophila subgenera and then the Drosophila subgenus is divided into the Hawaiian Drosophila and the repleta/virilis groups. Only the latter contain a duplicated hsp26. Comparing the gene organisation of the shsp cluster shows that all the Drosophila subgenus species contain the inversion. Putative heat shock elements (HSE) were found in the promoters of all the shsp and putative regulator elements for tissue specific expression were found in the promoter of hsp23, hsp27 and one of the hsp26 genes. hsp23 was found to be polymorphic for four non-synonymous changes that all lead to exchange of a Valine. The duplicated hsp26 gene in D. buzzatii (phsp26) was polymorphic for two non-synonymous changes. The allele frequencies of these variants were determined in nine D. buzzatii populations covering most of its distribution in Australia using high-resolution melting curves. The allele frequencies of one of the hsp23 variants showed a significant linear regression with longitude and the pooled frequency of the four Valine changes of hsp23 in the nine populations showed a significant linear regression with longitude and with a composite measure of climatic variables.

Direct and correlated responses to selection for longevity in Drosophila buzzatii

The possible associations between longevity, early fecundity, and stress-resistance traits were explored using artificial selection on longevity in a laboratory population of Drosophila buzzatii. Three replicated lines were selected for increased lifespan (L lines) and compared with the respective unselected controls (C lines) after the 14th generation of selection. Mean longevity exhibited a significant response to selection. The baseline mortality tended to decrease in the L lines and a negative correlated response to longevity selection was found for early fecundity. Egg-to-adult developmental time increased in L lines. Longevity selection increased stress resistance for both high and low temperatures, as measured by heat knockdown resistance and chill-coma recovery. Starvation resistance also tended to be higher in L than in C lines. The results obtained are consistent with the hypothesis of trade-offs between longevity and early fecundity, and also suggest a trade-off association between adult longevity and developmental time. Correlated selection responses were generally consistent with correlations among the traits previously inferred from altitudinal clines for longevity and stress-resistance phenotypes. © 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 97, 738–748.

Erratum: Bottlenecks, population differentiation and apparent selection at microsatellite loci in Australian Drosophila buzzatii

Correction to: Heredity (2009) 102, 389–401; doi:10.1038/hdy.2008.127 The authors have realized that an incorrect version of Table 3 was published. Please see the correct table below. The authors apologize for this error.

Local adaptation of stress related traits in Drosophila buzzatii and Drosophila simulans in spite of high gene flow

We addressed the question if local adaptation to a thermal gradient is possible in spite of a high gene flow among closely spaced populations of two species of Drosophila from the island of La Gomera (Canary Islands). Variation in multiple traits related to stress resistance in different life stages was measured in both species in flies collected from five localities at different altitudes and thereby with different climatic conditions. Based on microsatellite loci, the populations were not genetically differentiated. However, 18 of the 24 independent traits measured showed significant differentiation among populations of Drosophila buzzatii, but only nine of 25 for Drosophila simulans. This difference in the number of traits might reflect higher habitat specificity and thus higher potential for local adaptation of D. buzzatii than D. simulans. We found clinal variation, as some traits showed significant linear regressions on altitude, but more on altitude cubed.

Evolutionary dynamics and sites of illegitimate recombination revealed in the interspersion and sequence junctions of two nonhomologous satellite DNAs in cactophilic Drosophila species

Satellite DNA (satDNA) is a major component of genomes but relatively little is known about the fine-scale organization of unrelated satDNAs residing at the same chromosome location, and the sequence structure and dynamics of satDNA junctions. We studied the organization and sequence junctions of two nonhomologous satDNAs, pBuM and DBC-150, in three species from the neotropical Drosophila buzzatii cluster (repleta group). In situ hybridization to microchromosomes, interphase nuclei and extended DNA fibers showed frequent interspersion of the two satellites in D. gouveai, D. antonietae and, to a lesser extent, D. seriema. We isolated by PCR six pBuM x DBC-150 junctions: four are exclusive to D. gouveai and two are exclusive to D. antonietae. The six junction breakpoints occur at different positions within monomers, suggesting independent origin. Four junctions showed abrupt transitions between the two satellites, whereas two junctions showed a distinct 10 bp tandem duplication before the junction. Unlike pBuM, DBC-150 junction repeats are more variable than randomly cloned monomers and showed diagnostic features in common to a 3-monomer higher-order repeat seen in the sister species D. serido. The high levels of interspersion between pBuM and DBC-150 repeats suggest extensive rearrangements between the two satellites, maybe favored by specific features of the microchromosomes. Our interpretation is that the junctions evolved by multiples events of illegitimate recombination between nonhomologous satDNA repeats, with subsequent rounds of unequal crossing-over expanding the copy number of some of the junctions.

Cloning and sequencing of the breakpoint regions of inversion 5g fixed in Drosophila buzzatii

Chromosomal inversions are ubiquitous in Drosophila both as intraspecific polymorphisms and interspecific differences. Many gaps still remain in our understanding of the mechanisms that generate them. Previous work has shown that in Drosophila buzzatii, three polymorphic inversions were generated by ectopic recombination between copies of the transposon Galileo. In this study, we have characterized the breakpoint regions of inversion 5g, fixed in D. buzzatii and absent in Drosophila koepferae and other closely related species. A novel approach comprising four experimental steps was used. First, D. buzzatii BAC clones encompassing the breakpoints were identified and their ends sequenced. Then, breakpoint regions were mapped at high resolution in the Drosophila mojavensis genome sequence. Finally, breakpoint regions were isolated by polymerase chain reaction in D. buzzatii and D. koepferae and sequenced. Our aim was to shed light on the mechanism that generated inversion 5g and specifically to test for an implication of the transposon Galileo. No evidence implicates Galileo or other transposable elements in the origin of inversion 5g that was generated most likely by two independent breaks and non-homologous end-joining repair. Our results show that different inversion-generating mechanisms may coexist within the same lineage and suggest a hypothesis for the evolutionary time and mode of their operation.

Bottlenecks, population differentiation and apparent selection at microsatellite loci in Australian Drosophila buzzatii

Species colonizing new areas disjunct from their original habitat may be subject to novel selection pressures, and exhibit adaptive genetic changes. However, if colonization occurs through a small number of founders, the genetic composition of the colonized population may differ from that of the original population simply due to genetic drift. Disentangling the effects of founder drift and selection after colonization is crucial to understanding the adaptive process. Drosophila buzzatii colonized Australia some 600-700 generations ago, and spread rapidly over a wide geographical range. Genetic variation for 15 microsatellite loci in each of nine populations in eastern Australia was used to estimate the size of the bottleneck, and to determine if any of these microsatellites marked genomic regions subject to recent selection. We estimate that on its introduction to Australia, D. buzzatii went through a moderate bottleneck (approximately 30-40 founders). Linkage disequilibrium was common, both intrachromosomal and between loci on different chromosomes. Of the 15 loci, 2 showed evidence of selection, one exhibiting local adaptation in different populations and the other balancing selection. We conclude that linkage disequilibria may be far more common in natural populations than is generally assumed, and the loci apparently affected by selection may well be marking selection in large genome regions including many loci that are not necessarily closely linked.

Heat-induced hormesis in longevity as correlated response to thermal-stress selection in Drosophila buzzatii

Hormesis in longevity is a widespread phenomenon across the animal kingdom. It takes place when longevity is improved as an indirect effect of mild stress. We explored some possible evolutionary trajectories of hormesis in longevity in artificially selected lines of Drosophila buzzatii. The lines were bi-directionally selected for either knockdown resistance to heat stress (K −, K +) or chill-coma recovery (CCR −, CCR +, with the + and – signs indicating selection for decreased and increased tolerance, respectively). All K and CCR lines successfully diverged due to thermal-stress selection. The heat-inducible hormesis in longevity was substantial in both K − and CCR − females, whereas no hormesis was apparent for females in CCR +, K + and control lines. Among-line differences in longevity of non-heat-treated females disappeared after a heat-hardening treatment. Hormesis effects on the demographic senescence rate were sex-specific and consistently higher in the shorter-lived than in the longer-lived lines. Hormesis is an adaptive response, as its magnitude can evolutionary increase with stress-sensitivity.

Clinal variation in developmental time and viability, and the response to thermal treatments in two species of Drosophila

The present study first addressed the question of whether developmental time (DT) and viability (VT) vary clinally along latitudinal and altitudinal gradients in Drosophila buzzatii, an autochthonous specialist and the generalist invasive Drosophila melanogaster. Coincident and positive altitudinal clines across species and, direct and inverse latitudinal clines were observed for DT in D. melanogaster and D. buzzatii, respectively. Opposing latitudinal and altitudinal clines were detected for VT only in D. melanogaster. The patterns observed along altitudinal gradients prompted us to investigate whether flies living at lowland and highland environments may respond differentially to thermal treatments consisting of regimes of constant and alternating temperatures. Flies reared at higher mean temperature developed faster than at lower mean temperature in both species. By contrast, the response in VT differed greatly between species. Highland D. melanogaster were more viable than lowland regardless the treatment, whereas, in D. buzzatii, highland flies were more viable than lowland in alternating thermal regimes and the reverse was true in treatments of constant temperature. The results obtained suggest that thermal amplitude may be an important factor that should be considered in investigations of thermal adaptation.

A comparative study of competitive ability between two cactophilic species in their natural hosts

Abstract Competition is a major aspect of the ecology of insect communities exploiting ephemeral and fragmented resources. We analysed the effect of intraspecific (single species culture) and interspecific (mixed species culture) competition on larval viability, developmental time and wing length in the cactophilic Drosophila buzzatii and Drosophila koepferae (Diptera: Drosophilidae) reared in cultured media prepared with fermenting tissues of three common natural cactus hosts in nature at different densities. Our results show that all traits measured were affected by both intra‐ and interspecifc competition, although the effect of competition depended on the Drosophila species and the rearing cactus. In fact, flies tended to have a lower viability, shorter wing size and longer developmental time as a function of increasing density in single species culture in both D. buzzatii and D. koepferae (intraespecific competition). Besides, the performance of both species was seriously affected (shorter body size, slower developmental times, lower viability) by the presence of heterospecific competitors except in the case of D. koepferae reared in its primary host plant, Trichocereus terschekii. We also show that D. koepferae successfully utilized Opuntia quimilo, which is absent in most parts of its distribution range. We discuss the roles of intra‐ and interspecific competition as determinants of the relative abundance of these two species in the arid zones of Southern South America.

Sequence analysis, chromosomal distribution and long-range organization show that rapid turnover of new and old pBuM satellite DNA repeats leads to different patterns of variation in seven species of the Drosophila buzzatii cluster

We aimed to study patterns of variation and factors influencing the evolutionary dynamics of a satellite DNA, pBuM, in all seven Drosophila species from the buzzatii cluster (repleta group). We analyzed 117 alpha pBuM-1 (monomer length 190 bp) and 119 composite alpha/beta (370 bp) pBuM-2 repeats and determined the chromosome location and long-range organization on DNA fibers of major sequence variants. Such combined methodologies in the study of satDNAs have been used in very few organisms. In most species, concerted evolution is linked to high copy number of pBuM repeats. Species presenting low-abundance and scattered distributed pBuM repeats did not undergo concerted evolution and maintained part of the ancestral inter-repeat variability. The alpha and alpha/beta repeats colocalized in heterochromatic regions and were distributed on multiple chromosomes, with notable differences between species. High-resolution FISH revealed array sizes of a few kilobases to over 0.7 Mb and mutual arrangements of alpha and alpha/beta repeats along the same DNA fibers, but with considerable changes in the amount of each variant across species. From sequence, chromosomal and phylogenetic data, we could infer that homogenization and amplification events involved both new and ancestral pBuM variants. Altogether, the data on the structure and organization of the pBuM satDNA give insights into genome evolution including mechanisms that contribute to concerted evolution and diversification.

Molecular characterization of SSS139, a new satellite DNA family in sibling species of the Drosophila buzzatii cluster

We characterized sequences of a novel SSS139 RsaI satellite DNA family in Drosophila gouveai and Drosophila seriema, two members of the Drosophila buzzatii cluster (D. repleta group). The sequences were AT-rich (69%) with a monomer unit length of about 139 bp and contained two direct subrepeats of 14 bp and 16 bp, suggesting that it might have originated by the duplication of smaller sequences. Southern and dot-blot hybridization analyses also detected SSS139 in other Drosophila buzzatii cluster species (D. koepferae, D. antonietae, D. borborema and D. serido) but not in D. buzzatii. These results agree with the marginal phylogenetic position of D. buzzatii within the D. buzzatii cluster.

Wing morphology and fluctuating asymmetry depend on the host plant in cactophilicDrosophila

As in most insect groups, host plant shifts in cactophilic Drosophila represent environmental challenges as flies must adjust their developmental programme to the presence of different chemical compounds and/or to a microflora that may differ in the diversity and abundance of yeasts and bacteria. In this context, wing morphology provides an excellent opportunity to investigate the factors that may induce changes during development. In this work, we investigated phenotypic plasticity and developmental instability of wing morphology in flies on the cactophilic Drosophila buzzatii and Drosophila koepferae raised on alternative breeding substrates. We detected significant differences in wing size between and within species, and between flies reared on different cactus hosts. However, differences in wing shape between flies emerged from different cactus hosts were not significant either in D. buzzatii or in D. koepferae. Our results also showed that morphological responses involved the entire organ, as variation in size and shape correlated between different portions of the wing. Finally, we studied the effect of the rearing cactus host on developmental instability as measured by the degree of fluctuating asymmetry (FA). Levels of FA in wing size were significantly greater in flies of both species reared in non-preferred when compared with those reared in preferred host cacti. Our results are discussed in the framework of an integrative view aimed at investigating the relevance of host plant shifts in the evolution of the guild of cactophilic Drosophila species that diversified in South America.

The impact of genetic parental distance on developmental stability and fitness in Drosophila buzzatii

Measures of genetic parental distances (GPD) based on microsatellite loci (D(2) and IR), have been suggested to be better correlated with fitness than individual heterozygosity (H), as they contain information about past events of inbreeding or admixture. We investigated if GPD increased with increasing genetic divergence between parental populations in Drosophila buzzatii and if the measures indicate past events of admixture. Further we evaluated the relationship between GPD, fitness and fluctuating asymmetry (FA) of size and shape. We investigated three populations of Drosophila buzzati, from Argentina, Europe and Australia. From these populations two intraspecific hybridisation lines were made; one between the Argentinean and European populations, which have been separated 200 years and one between the populations from Argentina and Australia, which have been separated 80 years. By doing this we obtained hybrid progeny having different levels of GPD. We found that D(2) and H can be used as indicators of admixture when comparing hybrid individuals with their parentals. IR was not informative. Our results does not exclude the presence of genetic fitness correlations (GFC) over individuals with a broad fitness range from populations in equilibrium, but we doubt the presence of GFC using GPD measures in admixed populations. Shape FA could be a relevant measure for fitness, however, only when comparing populations, not at individual level.

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.