0000000001301567

AUTHOR

Anneli Hoikkala

Strength of sexual and postmating prezygotic barriers varies between sympatric populations with different histories and species abundances.

The impact of different reproductive barriers on species or population isolation may vary in different stages of speciation depending on evolutionary forces acting within species and through species' interactions. Genetic incompatibilities between interacting species are expected to reinforce prezygotic barriers in sympatric populations and lead to cascade reinforcement between conspecific populations living within and outside the areas of sympatry. We tested these predictions and studied whether and how the strength and target of reinforcement between Drosophila montana and Drosophila flavomontana vary between sympatric populations with different histories and species abundances. All barri…

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Myo-inositol as a main metabolite in overwintering flies: seasonal metabolomic profiles and cold stress tolerance in a northern drosophilid fly

SUMMARY Coping with seasonal changes in temperature is an important factor underlying the ability of insects to survive over the harsh winter conditions in the northern temperate zone, and only a few drosophilids have been able to colonize sub-polar habitats. Information on their winter physiology is needed as it may shed light on the adaptive mechanisms of overwintering when compared with abundant data on the thermal physiology of more southern species, such as Drosophila melanogaster. Here we report the first seasonal metabolite analysis in a Drosophila species. We traced changes in the cold tolerance and metabolomic profiles in adult Drosophila montana flies that were exposed to thermope…

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Effects of photoperiodically induced reproductive diapause and cold hardening on the cold tolerance of Drosophila montana

Coping with seasonal and daily variation in environmental conditions requires that organisms are able to adjust their reproduction and stress tolerance according to environmental conditions. Females of Drosophila montana populations have adapted to survive over the dark and cold winters at high latitudes and altitudes by spending this season in photoperiodically controlled reproductive diapause and reproducing only in spring/summer. The present study showed that flies of a northern population of this species are quite tolerant of low temperatures and show high seasonal and short-term plasticity in this trait. Culturing the flies in short day length (nearly all females in reproductive diapau…

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Two distinct genomic regions, harbouring the period and fruitless genes, affect male courtship song in Drosophila montana

Acoustic signals often have a significant role in pair formation and in species recognition. Determining the genetic basis of signal divergence will help to understand signal evolution by sexual selection and its role in the speciation process. An earlier study investigated quantitative trait locus for male courtship song carrier frequency (FRE) in Drosophila montana using microsatellite markers. We refined this study by adding to the linkage map markers for 10 candidate genes known to affect song production in Drosophila melanogaster. We also extended the analyses to additional song characters (pulse train length (PTL), pulse number (PN), interpulse interval, pulse length (PL) and cycle nu…

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Experimental introgression in Drosophila : Asymmetric postzygotic isolation associated with chromosomal inversions and an incompatibility locus on the X chromosome

Interspecific gene flow (introgression) is an important source of new genetic variation, but selection against it can reinforce reproductive barriers between interbreeding species. We used an experimental approach to trace the role of chromosomal inversions and incompatibility genes in preventing introgression between two partly sympatric Drosophila virilis group species, D. flavomontana and D. montana. We backcrossed F1 hybrid females from a cross between D. flavomontana female and D. montana male with the males of the parental species for two generations and sequenced pools of parental strains and their reciprocal 2nd generation backcross (BC2mon and BC2fla) females. Contrasting the obser…

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Transcriptional Differences between Diapausing and Non-Diapausing D. montana Females Reared under the Same Photoperiod and Temperature

Background A wide range of insects living at higher latitudes enter diapause at the end of the warm season, which increases their chances of survival through harsh winter conditions. In this study we used RNA sequencing to identify genes involved in adult reproductive diapause in a northern fly species, Drosophila montana. Both diapausing and non-diapausing flies were reared under a critical day length and temperature, where about half of the emerging females enter diapause enabling us to eliminate the effects of varying environmental conditions on gene expression patterns of the two types of female flies. Results RNA sequencing revealed large differences between gene expression patterns of…

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Direct and correlated responses to bi-directional selection on pre-adult development time in Drosophila montana.

Selection experiments offer an efficient way to study the evolvability of traits that play an important role in insects’ reproduction and/or survival and to trace correlations and trade-offs between them. We have exercised bi-directional selection on Drosophila montana flies’ pre-adult development time under constant light and temperature conditions for 10 generations and traced the indirect effects of this selection on females’ diapause induction under different day lengths, as well as on the body weight and cold tolerance of both sexes. Overall, selection was successful towards slow, but not towards fast development. However, all fast selection line replicates showed at the end of selecti…

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Latitudinal clines in the timing and temperature‐sensitivity of photoperiodic reproductive diapause in Drosophila montana

Reproductive diapause is a primary mechanism used by arthropods to synchronize their life cycle with seasonal changes in temperate regions. Our study species, Drosophila montana, represents the northern insect species where flies enter reproductive diapause under short day conditions and where the precise timing of diapause is crucial for both survival and offspring production. We have studied clinal variation in the critical day length for female diapause induction (CDL) and their overall susceptibility to enter diapause (diapause incidence), as well as the temperature sensitivity of these traits. The study was performed using multiple strains from four latitudinal clines of the species – …

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Changes in gene expression linked with adult reproductive diapause in a northern malt fly species: a candidate gene microarray study

Abstract Background Insect diapause is an important biological process which involves many life-history parameters important for survival and reproductive fitness at both individual and population level. Drosophila montana, a species of D. virilis group, has a profound photoperiodic reproductive diapause that enables the adult flies to survive through the harsh winter conditions of high latitudes and altitudes. We created a custom-made microarray for D. montana with 101 genes known to affect traits important in diapause, photoperiodism, reproductive behaviour, circadian clock and stress tolerance in model Drosophila species. This array gave us a chance to filter out genes showing expression…

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Cold adaptation drives population genomic divergence in the ecological specialist, Drosophila montana

Funding: UK Natural Environment Research Council (Grant Number(s): NE/L501852/1, NE/P000592/1); Academy of Finland (GrantNumber(s): 267244, 268214, 322980), Ella ja Georg Ehrnroothin Säätiö. Detecting signatures of ecological adaptation in comparative genomics is challenging, but analysing population samples with characterised geographic distributions, such as clinal variation, can help identify genes showing covariation with important ecological variation. Here, we analysed patterns of geographic variation in the cold-adapted species Drosophila montana across phenotypes, genotypes and environmental conditions and tested for signatures of cold adaptation in population genomic divergence. We…

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MALE COURTSHIP SONG AND FEMALE PREFERENCE VARIATION BETWEEN PHYLOGEOGRAPHICALLY DISTINCT POPULATIONS OF DROSOPHILA MONTANA

Understanding the variation within and between populations in important male mating traits and female preferences is crucial to theories concerning the origin of sexual isolation by coevolution or other processes. There have been surprisingly few studies on the extent of variation and covariation within and between populations, especially where the evolutionary relationships between populations are understood. Here we examine variation in female preferences and a sexually selected male song trait, the carrier frequency of the song, within and between populations from different phylogeographic clusters of Drosophila montana. Song is obligatory for successful mating in this species, and both …

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Adaptation and ecological speciation in seasonally varying environments at high latitudes: Drosophila virilis group

Living in high latitudes and altitudes sets specific requirements on species’ ability to forecast seasonal changes and to respond to them in an appropriate way. Adaptation into diverse environmental conditions can also lead to ecological speciation through habitat isolation or by inducing changes in traits that influence assortative mating. In this review, we explain how the unique time-measuring systems of Drosophila virilis group species have enabled the species to occupy high latitudes and how the traits involved in species reproduction and survival exhibit strong linkage with latitudinally varying photoperiodic and climatic conditions. We also describe variation in reproductive barriers…

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Adaptation to a seasonally varying environment: a strong latitudinal cline in reproductive diapause combined with high gene flow inDrosophila montana

Adaptation to seasonal changes in the northern hemisphere includes an ability to predict the forthcoming cold season from gradual changes in environmental cues early enough to prepare for the harsh winter conditions. The magnitude and speed of changes in these cues vary between the latitudes, which induces strong selection pressures for local adaptation. We studied adaptation to seasonal changes in Drosophila montana, a northern maltfly, by defining the photoperiodic conditions leading to adult reproductive diapause along a latitudinal cline in Finland and by measuring genetic differentiation and the amount of gene flow between the sampling sites with microsatellites. Our data revealed a cl…

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The extent of variation in male song, wing and genital characters among allopatric Drosophila montana populations.

Drosophila montana, a species of the Drosophila virilis group, has distributed around the northern hemisphere. Phylogeographic analyses of two North American and one Eurasian population of this species offer a good background for the studies on the extent of variation in phenotypic traits between populations as well as for tracing the selection pressures likely to play a role in character divergence. In the present paper, we studied variation in the male courtship song, wing and genital characters among flies from Colorado (USA), Vancouver (Canada) and Oulanka (Finland) populations. The phenotypic divergence among populations did not coincide with the extent of their genetic divergence, sug…

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The genetic architecture of sexually selected traits in two natural populations of Drosophila montana

The work was supported by the National Environment Research Council (grant NE/E015255/1 to MGR and RKB) and the Academy of Finland (project 132619 to AH). We investigated the genetic architecture of courtship song and cuticular hydrocarbon traits in two phygenetically distinct populations of Drosophila montana. To study natural variation in these two important traits, we analysed within-population crosses among individuals sampled from the wild. Hence, the genetic variation analysed should represent that available for natural and sexual selection to act upon. In contrast to previous between-population crosses in this species, no major quantitative trait loci (QTLs) were detected, perhaps be…

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Sequence variation in couch potato and its effects on life-history traits in a northern malt fly, Drosophila montana

Abstract Couch potato ( cpo ) has previously been connected to reproductive diapause in several insect species including Drosophila melanogaster , where it has been suggested to provide a link between the insulin signalling pathway and the hormonal control of diapause. In the first part of the study we sequenced nearly 3.6 kb of this gene in a northern Drosophila species ( Drosophila montana ) with a robust photoperiodically determined diapause and found several types of polymorphisms along the sequenced area. We also found variation among five Drosophila virilis group species in the length of the 5th exon of cpo and in the site of the stop codon at the end of this exon. The second part of …

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Northern Drosophila montana flies show variation both within and between cline populations in the critical day length evoking reproductive diapause

Reproductive diapause, and its correct timing, plays an important role in the life cycle of many insect species living in a seasonally varying environment at high latitudes. In the present paper we have documented variation in the critical day length (CDL) for adult reproductive diapause and the steepness of photoperiodic response curves (PPRCs) in seven clinal populations of Drosophila montana in Finland between the latitudes 61 and 67°N, paying special attention to variation in these traits within and between cline populations. The isofemale lines representing these populations showed a sharp transition from 0% to 100% in females' diapause incidence in the shortening day lengths, indicate…

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Strength of sexual and postmating prezygotic barriers varies between sympatric populations with different histories and species abundances

The impact of different reproductive barriers on species or population isolation may vary in different stages of speciation depending on evolutionary forces acting within species and through species’ interactions. Genetic incompatibilities between interacting species are expected to reinforce prezygotic barriers in sympatric populations and lead to cascade reinforcement between conspecific populations living within and outside the areas of sympatry. We tested these predictions and studied whether and how the strength and target of reinforcement between Drosophila montana and Drosophila flavomontana vary between sympatric populations with different histories and species abundances. All barri…

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sj-pdf-1-jbr-10.1177_0748730421997265 – Supplemental material for Nanda-Hamner Curves Show Huge Latitudinal Variation but No Circadian Components in Drosophila Montana Photoperiodism

Supplemental material, sj-pdf-1-jbr-10.1177_0748730421997265 for Nanda-Hamner Curves Show Huge Latitudinal Variation but No Circadian Components in Drosophila Montana Photoperiodism by Pekka Lankinen, Chedly Kastally and Anneli Hoikkala in Journal of Biological Rhythms

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A microsatellite linkage map forDrosophila montanashows large variation in recombination rates, and a courtship song trait maps to an area of low recombination

Current advances in genetic analysis are opening up our knowledge of the genetics of species differences, but challenges remain, particularly for out-bred natural populations. We constructed a microsatellite-based linkage map for two out-bred lines of Drosophila montana derived from divergent populations by taking advantage of the Drosophila virilis genome and available cytological maps of both species. Although the placement of markers was quite consistent with cytological predictions, the map indicated large heterogeneity in recombination rates along chromosomes. We also performed a quantitative trait locus (QTL) analysis on a courtship song character (carrier frequency), which differs be…

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Evolution of Complex Acoustic Signals in Drosophila Species

species produce complex acous-tic cues, so-called courtship songs, while pursuing a female.In most of the over 100 species studied so far (see thelist of these species in Hoikkala, 2005), such cues areproduced by wing vibration. Other mechanisms of songproduction include abdomen purring (Hoy, Hoikkala, &Kaneshiro, 1988) and rapid vibrations of the whole body(Ritchie & Gleason, 1995). The carrier frequency of songsproduced through any of these actions ranges from 150 to500Hz. A hitherto unknown mechanism enables males ofsome Hawaiian species to generate songs of up to 15,000Hz(Hoikkala, Hoy, & Kaneshiro, 1989).The structure of the courtship songs of closely relatedspecies often reflects phylo…

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Age-related decrease in male reproductive success and song quality in Drosophila montana

In the present paper, we have studied the effects of aging on male reproductive success and song quality in Drosophila montana .W e analyzed the reproductive success of wild-caught males at their normal breeding age during the mating season and after maintaining the males in laboratory from 1 to 5 months. In line with the mutation accumulation theory of aging, none of the factors affecting the reproductive success of wild-caught males during the mating season were related to male longevity. However, mating activity and progeny production of the males decreased with male age. Interestingly, there was no significant variation in progeny production between males at their normal breeding age, w…

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Reinforcement targets sexual or postmating prezygotic reproductive barriers depending on species abundance and population history

AbstractThe impact of different reproductive barriers on species or population isolation may vary in different stages of speciation depending on evolutionary forces acting within species and through species’ interactions. Genetic incompatibilities between interacting species are expected to reinforce prezygotic barriers in sympatric populations and create character displacement between conspecific populations living within and outside the area of sympatry. The outcome of reinforcement has been suggested to be affected by the strength of postzygotic barriers, the history of species coexistence, and the impact of species abundancies on females’ discrimination against heterospecific males. We …

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Sexual and postmating reproductive isolation between allopatric Drosophila montana populations suggest speciation potential

This work was funded by a European Commission Research Training Grant RTN2-2001-00049, the Centre of Excellence for Evolutionary Research at the University of Jyväskylä and a Marie Curie Initial Training Network, ‘Understanding the evolutionary origin of biological diversity’ (ITN-2008-213780 SPECIATION) Background: Widely distributed species with populations adapted to different environmental conditions can provide valuable opportunities for tracing the onset of reproductive incompatibilities and their role in the speciation process. Drosophila montana, a D. virilis group species found in high latitude boreal forests in Nearctic and Palearctic regions around the globe, could be an excellen…

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Phylogeographic patterns in Drosophila montana

The Drosophila virilis species group offers valuable opportunities for studying the roles of chromosomal re-arrangements and mating signals in speciation. The 13 species are divided into two subgroups, the montana and virilis 'phylads'. There is greater differentiation among species within the montana phylad in both karyotype and acoustic signals than exists among members of the virilis phylad. Drosophila montana is a divergent species which is included in the montana phylad. Here, we analyse the phylogeography of D. montana to provide a framework for understanding divergence of acoustic signals among populations. We analysed mitochondrial sequences corresponding to the cytochrome oxidase I…

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Photoperiodic regulation of life-history traits before and after eclosion: egg-to-adult development time, juvenile body weight and reproductive diapause in Drosophila montana

Photoperiod is the main environmental cue used by northern insects to predict the forthcoming seasonal changes and to adjust their life-history traits to fit these changes. We studied the effects of photoperiod on egg-to-adult development time, juvenile body mass and female reproductive diapause in two northern Drosophila montana populations with different patterns of voltinism. The most interesting findings were consistent between the populations: (1) when maintained before eclosion in short day conditions, representing early autumn, the flies developed faster and were lighter than when maintained in long day conditions, representing early summer, (2) photoperiodic time measurement is appa…

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Involvement of circadian oscillation(s) in the photoperiodic time measurement and the induction of reproductive diapause in a northern Drosophila species.

An ability to predict forthcoming changes in environmental conditions and get prepared for them in advance is crucial for the survival and reproduction of organisms living in a seasonally changing environment. We have studied the possible involvement of circadian oscillator(s) in the photoperiodic timer controlling seasonal responses by tracing Drosophila montana females' diapause induction in constant darkness and in a classical Nanda-Hamner experiment. Nearly all females developed ovaries in continuous darkness, which shows the direct development to be their default developmental pathway in the absence of photoperiods. In Nanda-Hamner experiment the females' diapause incidence was close t…

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Divergence and introgression among the virilis group of Drosophila

AbstractSpeciation with gene flow is now widely regarded as common. However, the frequency of introgression between recently diverged species and the evolutionary consequences of gene flow are still poorly understood. The virilis group of Drosophila contains around a dozen species that are geographically widespread and show varying levels of pre-zygotic and post-zygotic isolation. Here, we utilize de novo genome assemblies and whole-genome sequencing data to resolve phylogenetic relationships and describe patterns of introgression and divergence across the group. We suggest that the virilis group consists of three, rather than the traditional two, subgroups. We found evidence of pervasive p…

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Factors Affecting Male Song Evolution in Drosophila montana

D. montana (a species of the D. virilis group) has spread over the northern hemisphere, populations from different areas showing both genetic and phenotypic divergence. The males of this species produce an elaborate courtship song, which plays a major role both in species recognition and in intraspecific mate choice. The genetic architecture and physical constraints, as well as the importance of the signal for species recognition, set boundaries within which this signal can vary. Within these limits, courtship song parameters may change, depending on the males' physical condition and on the environment they inhabit. Females are likely to affect song evolution by exerting directional selecti…

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Selection for reproduction under short photoperiods changes diapause-associated traits and induces widespread genomic divergence.

The work has been supported by the Academyof Finland to A.H. (project 267244) and Natural Environment Research Council (NERC) funding (NE/J020818/1 to M.G.R.; NE/L501852/1 to R.A.W.W.). The incidence of reproductive diapause is a critical aspect of life history in overwintering insects from temperate regions. Much has been learned about the timing, physiology and genetics of diapause in a range of insects, but how the multiple changes involved in this and other photoperiodically regulated traits are inter-related is not well understood. We performed quasinatural selection on reproduction under short photoperiods in a northern fly species, Drosophila montana, to trace the effects of photoper…

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Microsatellite-based species identification method for Drosophila virilis group species

Species of the D. virilis group are widely used in evolutionary research, but the individuals of different species are difficult to distinguish from each other morphologically. We constructed a fast and easy microsatellite-based identification method for the species of the group occurring sympatrically in northern Europe. The neighbor joining tree based on 14 microsatellite loci also gave a good resolution of the species divergence pattern in the whole group.

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Additional file 1 of Multiple paths to cold tolerance: the role of environmental cues, morphological traits and the circadian clock gene vrille

Additional file 1: Table S1. Information on fly collecting sites and years, and the exact coordinates (latitude, longitude) and altitudes for each collecting site. Table S2. A List of 19 bioclimatic variables used in the PCA (WorldClim database v2.1, 2.5 min spatial resolutions; current data 1970–2000; Fick and Hijmans 2017; www.worldclim.org ). Table S3. 19 bioclimatic variables for each site were extracted from WorldClim database v2.1. Table S4. Principal components with their variance, cumulative variance and Eigenvalues. Table S5. Contributions (loadings) of the altitude and 19 bioclimatic variables on the Principal Component (PC). Table S6. The best-fit model for CCRT, CTmin, body colo…

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Inter- and intra-specific genomic divergence in Drosophila montana shows evidence for cold adaptation

This work was supported by the Academy of Finland to AH (projects 132619 and 267244) and to MK (projects 268214 and 272927) and NERC (UK) funding to MGR (grants NE/E015255/1 and NE/J020818/1) and PhD studentship to DJP (NE/I528634/1). The genomes of species that are ecological specialists will likely contain signatures of genomic adaptation to their niche. However, distinguishing genes related to ecological specialism from other sources of selection and more random changes is a challenge. Here we describe the genome of Drosophila montana, which is the most extremely cold-adapted Drosophila species. We use branch tests to identify genes showing accelerated divergence in contrasts between col…

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How consistent are the transcriptome changes associated with cold acclimation in two species of the Drosophila virilis group?

This work was financially support by a Marie Curie Initial Training Network grant, “Understanding the evolutionary origin of biological diversity” (ITN-2008–213780 SPECIATION), grants from the Academy of Finland to A.H. (project 132619) and M.K. (projects 268214 and 272927), a grant from NERC, UK to M.G.R. (grant NE/J020818/1), and NERC, UK PhD studentship to D.J.P. (NE/I528634/1). For many organisms the ability to cold acclimate with the onset of seasonal cold has major implications for their fitness. In insects, where this ability is widespread, the physiological changes associated with increased cold tolerance have been well studied. Despite this, little work has been done to trace chang…

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When are vomiting males attractive? Sexual selection on condition-dependent nuptial feeding in Drosophila subobscura

Nuptial gifts are any nutritious items or inedible tokens transferred from the male to the female as a part of courtship or copulation. Although nuptial gift donation has been studied in a variety of taxa, this behavior has been largely overlooked in Drosophila. We studied nuptial feeding in Drosophila subobscura, where the gift is a regurgitated drop of liquid, in order to examine the importance of this behavior for male mating success and female fecundity. We varied male and female condition by dietary restriction to assess any condition dependence of male nuptial feeding ability and female feeding behavior and mate discrimination. Our results show that there was directional selection for…

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Seasonal gene expression kinetics between diapause phases in Drosophila virilis group species and overwintering differences between diapausing and non-diapausing females.

AbstractMost northern insect species experience a period of developmental arrest, diapause, which enables them to survive over the winter and postpone reproduction until favorable conditions. We studied the timing of reproductive diapause and its long-term effects on the cold tolerance of Drosophila montana, D. littoralis and D. ezoana females in seasonally varying environmental conditions. At the same time we traced expression levels of 219 genes in D. montana using a custom-made microarray. We show that the seasonal switch to reproductive diapause occurs over a short time period and that overwintering in reproductive diapause has long-lasting effects on cold tolerance. Some genes, such as…

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Localization of quantitative trait loci for diapause and other photoperiodically regulated life history traits important in adaptation to seasonally varying environments.

Seasonally changing environments at high latitudes present great challenges for the reproduction and survival of insects, and photoperiodic cues play an important role in helping them to synchronize their life cycle with prevalent and forthcoming conditions. We have mapped quantitative trait loci (QTL) responsible for the photoperiodic regulation of four life history traits, female reproductive diapause, cold tolerance, egg-to-eclosion development time and juvenile body weight in Drosophila montana strains from different latitudes in Canada and Finland. The F2 progeny of the cross was reared under a single photoperiod (LD cycle 16:8), which the flies from the Canadian population interpret a…

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Multiple paths to cold tolerance: the role of environmental cues, morphological traits and the circadian clock gene vrille

AbstractBackgroundTracing the association between insect cold tolerance and latitudinally and locally varying environmental conditions, as well as key morphological traits and molecular mechanisms, is essential for understanding the processes involved in adaptation. We explored these issues in two closely-related species, Drosophila montana and Drosophila flavomontana, originating from diverse climatic locations across several latitudes on the coastal and mountainous regions of North America. We also investigated the association between sequence variation in one of the key circadian clock genes, vrille, and cold tolerance in both species. Finally, we studied the impact of vrille on fly cold…

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Cuticular hydrocarbons of Drosophila montana: geographic variation, sexual dimorphism and potential roles as pheromones.

Abstract Sexual selection within populations can play an important role in speciation when divergence in mating signals and their corresponding preferences occur along different coevolutionary trajectories in different populations. In insects, one potential target of sexual selection is the blend of cuticular hydrocarbons (CHCs), which often show intra- and interspecific variation, sexual dimorphism and may act as pheromones. In Drosophila montana, a cold-adapted, circumboreal member of the Drosophila virilis species group, flies from different populations have been found to show significant premating isolation as well as variation in male mating signal (song) and female preference. While t…

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Cold tolerance and cold-induced modulation of gene expression in two Drosophila virilis group species with different distributions

Abstract The importance of high and low temperature tolerance in adaptation to changing environmental conditions has evoked new interest in modulations in gene expression and metabolism linked with stress tolerance. We investigated the effects of rapid cold hardening and cold acclimatization on the chill coma recovery times of two Drosophila virilis group species, Drosophila montana and D. virilis, with different distributions and utilized a candidate gene approach to trace changes in their gene expression during and after the cold treatments. The study showed that cold acclimatization clearly decreases chill coma recovery times in both species, whereas rapid cold hardening did not have a s…

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Sexual conflict over the duration of copulation in Drosophila montana: why is longer better?

Background Conflicts of interest between the sexes are increasingly recognized as an engine driving the (co-)evolution of reproductive traits. The reproductive behaviour of Drosophila montana suggests the occurrence of sexual conflict over the duration of copulation. During the last stages of copulation, females vigorously attempt to dislodge the mounting male, while males struggle to maintain genital contact and often successfully extend copulations far beyond the females' preferred duration. Results By preventing female resistance, we show that females make a substantial contribution towards shortening copulations. We staged matings under different sex ratio conditions, and provide eviden…

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Nanda-Hamner Curves Show Huge Latitudinal Variation but No Circadian Components in Drosophila Montana Photoperiodism

Insect species with a wide distribution offer a great opportunity to trace latitudinal variation in the photoperiodic regulation of traits important in reproduction and stress tolerances. We measured this variation in the photoperiodic time-measuring system underlying reproductive diapause in Drosophila montana, using a Nanda-Hamner (NH) protocol. None of the study strains showed diel rhythmicity in female diapause proportions under a constant day length (12 h) and varying night lengths in photoperiods ranging from 16 to 84 h at 16°C. In the northernmost strains (above 55°N), nearly all females entered diapause under all photoperiods and about half of them even in continuous darkness, whil…

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Multiple quantitative trait loci influence intra-specific variation in genital morphology between phylogenetically distinct lines of Drosophila montana

The evolution of animal genitalia has gained renewed interest because of their potential roles during sexual selection and early stages of species formation. Although central to understanding the evolutionary process, knowledge of the genetic basis of natural variation in genital morphology is limited to a very few species. Using an outbred cross between phylogenetically distinct lines of Drosophila montana, we characterized quantitative trait loci (QTLs) affecting the size and shape of the distiphallus, a prominent part of the male intromittent organ. Our microsatellite-based linkage analysis shows that intra-specific variation in the distiphallus involves several QTLs of largely additive …

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Diapause affects cuticular hydrocarbon composition and mating behavior of both sexes inDrosophila montana

Environmental cues, mainly photoperiod and temperature, are known to control female adult reproductive diapause in several insect species. Diapause enhances female survival during adverse conditions and postpones progeny production to the favorable season. Male diapause (a reversible inability to inseminate receptive females) has been studied much less than female diapause. However, if the males maximized their chances to fertilize females while minimizing their energy expenditure, they would be expected to be in diapause at the same time as females. We investigated Drosophila montana male mating behavior under short‐day conditions that induce diapause in females and found the males to be r…

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Photoperiodic regulation of cold tolerance and expression levels of regucalcin gene in Drosophila montana

Temperature-induced plasticity of cold tolerance has been reported in many insect species, but cold tolerance can also be affected by changes in day (or night) length. In the present study we elucidate the direct and indirect effects of photoperiod on the cold tolerance of females of two Drosophila montana strains--one which possesses a robust photoperiodic diapause and another which does not. In the diapause-strain the time needed for recovery from chill coma showed a positive correlation with day length, but diapause itself played only a minor role in photoperiodic acclimation. The strain that was not able to enter to diapause as a response to day length also lacked photoperiodic cold acc…

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Signals of demographic expansion in Drosophila virilis

Background. The pattern of genetic variation within and among populations of a species is strongly affected by its phylogeographic history. Analyses based on putatively neutral markers provide data from which past events, such as population expansions and colonizations, can be inferred. Drosophila virilis is a cosmopolitan species belonging to the virilis group, where divergence times between different phylads go back to the early Miocene. We analysed mitochondrial DNA sequence variation among 35 Drosophila virilis strains covering the species' range in order to detect demographic events that could be used to understand the present characteristics of the species, as well as its differences …

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What do we need to know about speciation?

Speciation has been a major focus of evolutionary biology research in recent years, with many important advances. However, some of the traditional organising principles of the subject area no longer provide a satisfactory framework, such as the classification of speciation mechanisms by geographical context into allopatric, parapatric and sympatry classes. Therefore, we have asked where speciation research should be directed in the coming years. Here, we present a distillation of questions about the mechanisms of speciation, the genetic basis of speciation and the relationship between speciation and diversity. Our list of topics is not exhaustive; rather we aim to promote discussion on rese…

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Effect of temperature on the duration of sensitive period and on the number of photoperiodic cycles required for the induction of reproductive diapause in Drosophila montana

Abstract Correct timing of the induction of photoperiodic reproductive diapause has been found to play an important role in the life cycle of several northern insect species. However, even when the environmental conditions are favourable for diapause, the switch to diapause can only take place when the females are in a proper developmental and physiological stage, referred to as the sensitive period (SP) for diapause. We have previously shown that in a northern fly species, Drosophila montana, the developmental pathway of the ovaries (direct maturation vs. diapause) is determined by photoperiodic cues that the females receive after eclosion. Here, we have studied the effects of temperature …

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Circadian clock of Drosophila montana is adapted to high variation in summer day lengths and temperatures prevailing at high latitudes

Photoperiodic regulation of the circadian rhythms in insect locomotor activity has been studied in several species, but seasonal entrainment of these rhythms is still poorly understood. We have traced the entrainment of activity rhythm of northern Drosophila montana flies in a climate chamber mimicking the photoperiods and day and night temperatures that the flies encounter in northern Finland during the summer. The experiment was started by transferring freshly emerged females into the chamber in early and late summer conditions to obtain both non-diapausing and diapausing females for the studies. The locomotor activity of the females and daily changes in the expression levels of two core …

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Plasticity in Photoperiodism : Drosophila montana Females Have a Life-Long Ability to Switch From Reproduction to Diapause

Photoperiodic reproductive diapause is an essential part of female life cycle in several insect species living on high latitudes, where overwintering in reproductive stage involves high risks for survival and progeny production. The sensitive period (SP), during which photoperiodic cues can trigger the switch from direct development to diapause, can last from a few hours or days after emergence to the entire life span of females. Moreover, in some species, sexually mature females can enter post-reproductive diapause as a response to decreasing day length and/or temperature. We studied the duration of SP for diapause induction and the females’ ability to enter post-reproductive diapause at s…

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Intraspecific variation in mating behaviour does not cause sexual isolation between Drosophila virilis strains

There is increasing evidence that mate recognition systems vary within species. We studied the courtship rituals of five Drosophila virilis strains to investigate this intraspecific variation. Males that actively and persistently courted the female without paying much attention to the female's behaviour were the most successful at copulating. Intraspecific variation in mating rituals was high, but not as high as at the species level and it was not high enough to cause sexual isolation between the strains. The differences in courtship interactions of the strains reflected their geographical origin. Males from one of the strains did not produce audible song during their courtship, but still m…

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Additional file 1 of Multiple paths to cold tolerance: the role of environmental cues, morphological traits and the circadian clock gene vrille

Additional file 1: Table S1. Information on fly collecting sites and years, and the exact coordinates (latitude, longitude) and altitudes for each collecting site. Table S2. A List of 19 bioclimatic variables used in the PCA (WorldClim database v2.1, 2.5 min spatial resolutions; current data 1970–2000; Fick and Hijmans 2017; www.worldclim.org ). Table S3. 19 bioclimatic variables for each site were extracted from WorldClim database v2.1. Table S4. Principal components with their variance, cumulative variance and Eigenvalues. Table S5. Contributions (loadings) of the altitude and 19 bioclimatic variables on the Principal Component (PC). Table S6. The best-fit model for CCRT, CTmin, body colo…

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Sexual selection on song and cuticular hydrocarbons in two distinct populations ofDrosophila montana

Sexual selection has the potential to contribute to population divergence and speciation. Most studies of sexual selection in Drosophila have concentrated on a single signaling modality, usually either courtship song or cuticular hydrocarbons (CHCs), which can act as contact pheromones. We have examined the relationship between both signal types and reproductive success using F1–3 offspring of wild-collected flies, raised in the lab. We used two populations of the Holarctic species Drosophila montana that represent different phylogeographic clades that have been separate for ca. 0.5 million years (MY), and differ to some extent in both traits. Here, we characterize the nature and identify t…

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Flies in the north: locomotor behavior and clock neuron organization of Drosophila montana.

The circadian clock plays an important role in adaptation in time and space by synchronizing changes in physiological, developmental, and behavioral traits of organisms with daily and seasonal changes in their environment. We have studied some features of the circadian activity and clock organization in a northern Drosophila species, Drosophila montana, at both the phenotypic and the neuronal levels. In the first part of the study, we monitored the entrained and free-running locomotor activity rhythms of females in different light-dark and temperature regimes. These studies showed that D. montana flies completely lack the morning activity component typical to more southern Drosophila speci…

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sj-pdf-1-jbr-10.1177_0748730421997265 – Supplemental material for Nanda-Hamner Curves Show Huge Latitudinal Variation but No Circadian Components in Drosophila Montana Photoperiodism

Supplemental material, sj-pdf-1-jbr-10.1177_0748730421997265 for Nanda-Hamner Curves Show Huge Latitudinal Variation but No Circadian Components in Drosophila Montana Photoperiodism by Pekka Lankinen, Chedly Kastally and Anneli Hoikkala in Journal of Biological Rhythms

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Reproductive isolation among allopatric Drosophila montana populations

An outstanding goal in speciation research is to trace the mode and tempo of the evolution of barriers to gene flow. Such research benefits from studying incipient speciation, in which speciation between populations has not yet occurred, but where multiple potential mechanisms of reproductive isolation (RI: i.e., premating, postmating-prezygotic (PMPZ), and postzygotic barriers) may act. We used such a system to investigate these barriers among allopatric populations of Drosophila montana. In all heteropopulation crosses we found premating (sexual) isolation, which was either symmetric or asymmetric depending on the population pair compared. Postmating isolation was particularly strong in c…

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Data from: Localization of QTL for diapause and other photoperiodically regulated life-history traits important in adaptation to seasonally varying environments

Seasonally changing environments at high latitudes present great challenges for the reproduction and survival of insects, and photoperiodic cues play an important role in helping them to synchronize their life cycle with prevalent and forthcoming conditions. We have mapped quantitative trait loci (QTL) responsible for the photoperiodic regulation of four life history traits, female reproductive diapause, cold tolerance, egg-to-eclosion development time and juvenile body weight in Drosophila montana strains from different latitudes in Canada and Finland. The F2 progeny of the cross was reared under a single photoperiod (LD cycle 16:8), which the flies from the Canadian population interpret a…

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Data from: Inter- and intra-specific genomic divergence in Drosophila montana shows evidence for cold adaptation

D. montana gff fileGenome annotation file for D. montana genome (Accession number: LUVX00000000)D.mont_freeze_v1.4.gff.txt

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Data from: Two distinct genomic regions, harbouring the period and fruitless genes, affect male courtship song in Drosophila montana

Acoustic signals often play a significant role in pair formation and in species recognition. Determining the genetic basis of signal divergence will help to understand signal evolution by sexual selection and its role in the speciation process. An earlier study investigated QTL for male courtship song carrier frequency in Drosophila montana using microsatellite markers. We refined this study by adding to the linkage map markers for ten candidate genes known to affect song production in D. melanogaster. We also extended the analyses to additional song characters (pulse train length, pulse number, interpulse interval, pulse length and cycle number). Our results indicate that loci in two diffe…

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Data from: The genetic architecture of sexually selected traits in two natural populations of Drosophila montana

We investigated the genetic architecture of courtship song and cuticular hydrocarbon traits in two phygenetically distinct populations of Drosophila montana. In order to study natural variation in these two important traits, we analysed within-population crosses among individuals sampled from the wild. Hence, the genetic variation analysed should represent that available for natural and sexual selection to act upon. In contrast to previous between-population crosses in this species, no major QTLs were detected, perhaps because the between-population QTL were due to fixed differences between the populations. Partitioning the trait variation to chromosomes suggested a broadly polygenic geneti…

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Data from: Sexual selection on song and cuticular hydrocarbons in two distinct populations of Drosophila montana

Sexual selection has the potential to contribute to population divergence and speciation. Most studies of sexual selection in Drosophila have concentrated on a single signaling modality, usually either courtship song or cuticular hydrocarbons (CHCs), which can act as contact pheromones. We have examined the relationship between both signal types and reproductive success using F1-3 offspring of wild-collected flies, raised in the lab. We used two populations of the Holarctic species Drosophila montana, that represent different phylogeographic clades which have been separate for ca. 0.5 MY, and which differ to some extent in both traits. Here we characterise the nature and identify the target…

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Data from: Strength of sexual and postmating prezygotic barriers varies between sympatric populations with different histories and species abundances

The impact of different reproductive barriers on species or population isolation may vary in different stages of speciation depending on evolutionary forces acting within species and through species’ interactions. Genetic incompatibilities between interacting species are expected to reinforce prezygotic barriers in sympatric populations and lead to cascade reinforcement between conspecific populations living within and outside the areas of sympatry. We tested these predictions and studied whether and how the strength and target of reinforcement between Drosophila montana and Drosophila flavomontana vary between sympatric populations with different histories and species abundances. All barri…

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Data from: Multiple quantitative trait loci influence intra-specific variation in genital morphology between phylogenetically distinct lines of Drosophila montana

The evolution of animal genitalia has gained renewed interest, because of their potential roles during sexual selection and early stages of species formation. Although central to understanding the evolutionary process, knowledge of the genetic basis of natural variation in genital morphology is limited to a very few species. Using an out-bred cross between phylogenetically distinct lines of Drosophila montana, we characterized quantitative trait loci (QTLs) affecting the size and shape of the distiphallus, a prominent part of the male intromittent organ. Our microsatellite-based linkage analysis shows that intra-specific variation of the distiphallus involves several QTLs of largely additiv…

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Data from: Reproductive isolation among allopatric Drosophila montana populations

An outstanding goal in speciation research is to trace the mode and tempo of the evolution of barriers to gene flow. Such research benefits from studying incipient speciation, in which speciation between populations has not yet occurred but where multiple potential mechanisms of reproductive isolation (RI: i.e. premating, postmating-prezygotic (PMPZ), and postzygotic barriers) may act. We used such a system to investigate these barriers among allopatric populations of Drosophila montana. In all heteropopulation crosses we found premating (sexual) isolation which was either symmetric or asymmetric depending on the population pair compared. Postmating isolation was particularly strong in cros…

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Sexual and postmating reproductive isolation between allopatric Drosophila montana populations suggest speciation potential

Background - Widely distributed species with populations adapted to different environmental conditions can provide valuable opportunities for tracing the onset of reproductive incompatibilities and their role in the speciation process. Drosophila montana, a D. virilis group species found in high latitude boreal forests in Nearctic and Palearctic regions around the globe, could be an excellent model system for studying the early stages of speciation, as a wealth of information concerning this species' ecology, mating system, life history, genetics and phylogeography is available. However, reproductive barriers between populations have hereto not been investigated. Results - We report both pr…

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Changes in gene expression linked with adult reproductive diapause in a northern malt fly species: a candidate gene microarray study

Background: Insect diapause is an important biological process which involves many life-history parameters important for survival and reproductive fitness at both individual and population level. Drosophila montana, a species of D. virilis group, has a profound photoperiodic reproductive diapause that enables the adult flies to survive through the harsh winter conditions of high latitudes and altitudes. We created a custom-made microarray for D. montana with 101 genes known to affect traits important in diapause, photoperiodism, reproductive behaviour, circadian clock and stress tolerance in model Drosophila species. This array gave us a chance to filter out genes showing expression changes…

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