The influence of induced mutagenesis on reproductive behavior of Drosophila subobscura and the role of sexual selection in relationto the level of mutational load

Prema teorijskim očekivanjima seksualna selekcija može smanjivati mutaciona opterećenja u populacijama. Mužjaci koji su boljeg genetičkog kvaliteta bi trebalo da budu uspešniji u parenju u poređenju sa mužjacima lošijeg genetičkog kvaliteta. Na taj način ženke, prilikom izbora partnera za parenje, mogu da eliminišu one mužjake koji nose štetne alele, i da smanje mogućnost prenošenja štetnih alela u sledeću generaciju. Seksualna selekcija će redukovati mutaciona opterećenja i povećavati adaptivnu vrednost populacije, ako je većina mutacija štetna i za adaptivnu vrednost i za osobine koje utiču na uspešnost mužjaka u parenju. Istraživanja poslednjih godina su dala rezultate koji idu u prilog ovoj teoriji, ali i oprečne rezultate. Ovakva kontradiktornost proizilazi iz kompleksnosti problematike usled različitih reproduktivnih strategija vrsta, kao i različitih reproduktivnih strategija mužjaka i ženki. Takođe, osobine koje utiču na komponente adaptivne vrednosti, kao i na uspešnost u parenju, su veoma složene, pod determinacijom velikog broja gena, i u čijoj osnovi leže kompleksne genske interakcije. Iako se kao osobine koje su cilj seksualne selekcije najčešće analiziraju sekundarne polne karakteristike, i mnoge druge osobine određuju atraktivnost mužjaka. To su i morfološke, fiziološke, ponašajne i druge osobine koje utiču na uspešnost u parenju, a mogu se nazvati osobinama koje su zavisne od opšteg stanja mužjaka (eng. condition-dependent traits)...According to theoretical predictions sexual selection can reduce mutational load in populations. Males of good genetic quality should be more successful in matings, compared to the males of low genetic quality. In this way females, through the choice of mating partners, can eliminate males carriers of deleterious alleles, and reduce transmission of deleterious alleles to the next generation. Sexual selection will purge mutational load and increase fitness if most mutations were deleterious to both nonsexual fitness and condition-dependent traits affecting mating success. Studies in recent years have yielded results that support this theory, but opposite results are obtained, as well. This contradiction arises from different reproductive strategies of species, and different reproductive strategies of males and females. Traits that affect fitness components, as well as mating success, are under polygenic determination, with very complex gene interactions. Although secondary sexual traits were usually analyzed as target of sexual selection, many other traits can determine male attractiveness. Traits such as morphological, physiological, behavior and other, affecting mating success, can be termed as condition-dependent traits. In order to test this hypothesis, that sexual selection can reduce mutational load, two experimental groups were formed from the same population genetic pool of Drosophila subobscura, which differed in the level of mutational load. Mutations were induced in one group, using 30Gy ionizing radiation. The effect of sexual selection in reduction of mutational load was measured through different components of male mating behavior (courtship occurrence, courtship latency, courtship intensity, mating occurrence,latency to mating and duration of mating)...

The influence of induced mutagenesis on reproductive behavior of Drosophila subobscura and the role of sexual selection in relationto the level of mutational load

Prema teorijskim očekivanjima seksualna selekcija može smanjivati mutaciona opterećenja u populacijama. Mužjaci koji su boljeg genetičkog kvaliteta bi trebalo da budu uspešniji u parenju u poređenju sa mužjacima lošijeg genetičkog kvaliteta. Na taj način ženke, prilikom izbora partnera za parenje, mogu da eliminišu one mužjake koji nose štetne alele, i da smanje mogućnost prenošenja štetnih alela u sledeću generaciju. Seksualna selekcija će redukovati mutaciona opterećenja i povećavati adaptivnu vrednost populacije, ako je većina mutacija štetna i za adaptivnu vrednost i za osobine koje utiču na uspešnost mužjaka u parenju. Istraživanja poslednjih godina su dala rezultate koji idu u prilog ovoj teoriji, ali i oprečne rezultate. Ovakva kontradiktornost proizilazi iz kompleksnosti problematike usled različitih reproduktivnih strategija vrsta, kao i različitih reproduktivnih strategija mužjaka i ženki. Takođe, osobine koje utiču na komponente adaptivne vrednosti, kao i na uspešnost u parenju, su veoma složene, pod determinacijom velikog broja gena, i u čijoj osnovi leže kompleksne genske interakcije. Iako se kao osobine koje su cilj seksualne selekcije najčešće analiziraju sekundarne polne karakteristike, i mnoge druge osobine određuju atraktivnost mužjaka. To su i morfološke, fiziološke, ponašajne i druge osobine koje utiču na uspešnost u parenju, a mogu se nazvati osobinama koje su zavisne od opšteg stanja mužjaka (eng. condition-dependent traits)...According to theoretical predictions sexual selection can reduce mutational load in populations. Males of good genetic quality should be more successful in matings, compared to the males of low genetic quality. In this way females, through the choice of mating partners, can eliminate males carriers of deleterious alleles, and reduce transmission of deleterious alleles to the next generation. Sexual selection will purge mutational load and increase fitness if most mutations were deleterious to both nonsexual fitness and condition-dependent traits affecting mating success. Studies in recent years have yielded results that support this theory, but opposite results are obtained, as well. This contradiction arises from different reproductive strategies of species, and different reproductive strategies of males and females. Traits that affect fitness components, as well as mating success, are under polygenic determination, with very complex gene interactions. Although secondary sexual traits were usually analyzed as target of sexual selection, many other traits can determine male attractiveness. Traits such as morphological, physiological, behavior and other, affecting mating success, can be termed as condition-dependent traits. In order to test this hypothesis, that sexual selection can reduce mutational load, two experimental groups were formed from the same population genetic pool of Drosophila subobscura, which differed in the level of mutational load. Mutations were induced in one group, using 30Gy ionizing radiation. The effect of sexual selection in reduction of mutational load was measured through different components of male mating behavior (courtship occurrence, courtship latency, courtship intensity, mating occurrence,latency to mating and duration of mating)...

Sexual selection can reduce mutational load in Drosophila subobscura

According to theoretical predictions sexual selection can reduce mutational load through male mating success. Males of good genetic quality should be more successful in matings, compared to the males of low genetic quality, thus in this way females can prevent deleterious alleles to be transmitted to the next generation. We tested this hypothesis through set up of two experimental groups from same genetic pool, where in one group genetic quality was manipulated by ionizing radiation. Within each group opportunity for choosing mates was imposed: males and females had no choice or had multiple choice. Mutational load was measured through the variability of different fitness components: fecundity and egg-to-adult viability. Our results indicate that sexual selection can reduce mutational load, only for fecundity. Group with the presence of female choice exhibited higher fecundity than group in which sexual selection was experimentally eliminated, but only in 'irradiated' group. There was no overall difference in egg-to-adult viability between different sexual selection regimes in any of the group. It should be considered that sexual selection can cause sexual conflict, and potential opposite effects of sexual selection and sexual conflict on fitness. Genetic structure of populations, in terms of the level of mutational load, is an important factor which can determinate the role of sexual selection.Prema teorijskim očekivanjima seksualna selekcija može smanjivati mutaciona opterećenja u populacijama, preko uspešnosti mužjaka u parenju. Mužjaci koji su boljeg genetičkog kvaliteta bi trebalo da budu uspešniji u parenju od mužjaka lošijeg genetičkog kvaliteta. Na taj način bi ženke mogle da smanje prenošenje štetnih alela u sledeću generaciju. Ova hipoteza je testirana uspostavljanjem dve eksperimentalne grupe od istog genetičkog pula jedinki, pri čemu su u jednoj mutacije indukovane jonizujućim zračenjem. Unutar svake grupe je nametnuta mogućnost izbora u parenju: mužjaci i ženke nisu imali izbor, ili su imali višestruki izbor. Mutaciona opterećenja su merena preko dve komponente adaptivne vrednosti: fekunditeta i preživljavanja od stadijuma jaja do adulta. Dobijeni rezultati ukazuju da seksualna selekcija može imati uticaj na smanjivanje mutacionih opterećenja ali samo za fekunditet. U prisustvu seksualne selekcije su vrednosti fekunditeta bile veće u odnosu na vrednosti koje su dobijene kad je seksualna selekcija eksperimentalno uklonjena, ali samo u okviru 'ozračene' grupe. Razlike u preživljavanju od jaja do adulta između različitih režima seksualne selekcije nisu dobijene ni u jednoj od grupa. Kako seksualna selekcija često uzrokuje i seksualni konflikt, treba uzeti u obzir njihovo potencijalno delovanje u suprotnim pravcima na adaptivnu vrednost. Genetička struktura populacija, koja podrazumeva i veličinu mutacionih opterećenja, može biti važan faktor od koga će i zavisiti delovanje seksualne selekcija na adaptivnu vrednost.Projekat ministarstva br. 17301

Synergistic effect of environmental and genomic stress on wing size of drosophila subobscura

Growing anthropogenic influence on every aspect of environment arise important issues regarding the ability of populations and species to adapt to variant pressures. Lead is one of the most present contaminants in the environment with detrimental influence on organisms and populations. In combination with genomic stress, lead may act synergistically, leading to reduction in adaptive values. We sampled two Drosophila subobscura populations, from ecologically different habitats and established differences in genetic backgrounds and population histories. In order to establish different levels of genome heterozygosity, series of intra-line, intra-population and between population crosses were made. The progeny was reared on a standard Drosophila medium and a medium with 200μg/mL of lead acetate and right wing of approximately 4000 individuals was used for geometric morphometric analysis of wing size. Results showed that lead significantly reduces wing size and that magnitude of this reduction is dependent on genetic background, indicating synergistic effect of genomic and environmental stress. There is also an indication of strong female origin influence on the outcome of hybridization when source of environmental stress is lead. Our results showed that the genetic structure of populations is of great importance for population fitness in anthropogenic induced stressful conditions. Further studies of synergistic effect of genetic and environmental stress are needed, as well as studies of its outcome in natural populations.Genetika (2016), 48(3): 1039-105

Local adaptation at fine spatial scale through chromosomal inversions and mito-nuclear epistasis: Findings in Drosophila subobscura (Diptera: Drosophilidae)

To explore local adaptation in wild populations at a fine spatial scale we characterized the genetic variability of eight closely located populations of Drosophila subobscura and its associations with microhabitat environmental conditions. Three different genetic markers were assessed: chromosomal inversions, a SNP of mitochondrial ND5 gene and nuclear microsatellites. Population genetic analyses of chromosomal variability revealed significant genetic differentiation between these populations. Gene arrangement frequencies on the E chromosome contributed most to these differences. We also investigated role of mito-nuclear epistasis in mitochondrial genome differentiation and revealed weak linkage disequilibrium (LD) exclusively between O3+4 inversion arrangement and mitochondrial DNA haplotype I in two populations. In addition, the trend in the LD between OST chromosomal arrangement and haplotype II was general in the total sample. Microsatellite analysis revealed an absence of stochastic processes, like census reduction, upon population differentiation. Only a small amount of the genetic variation is related to geographic distance, while most (97%) is attributable to other factors and in some degree to microhabitat variables (temperature, humidity). The analysis of these factors revealed they effect inversion arrangement frequencies, especially E1+2+9, EST and OST. Even though this model organism is known for its high mobility and mostly large effective population size, the results presented here reveal that local adaptations can occur even at a small spatial scale. We propose that locally adapted alleles within chromosomal inversions, as well as joint selective pressures acting on mitochondrial and nuclear genomes, are responsible for the observed adaptation to microhabitat conditions

Sexual selection can reduce mutational load in Drosophila subobscura

According to theoretical predictions sexual selection can reduce mutational load through male mating success. Males of good genetic quality should be more successful in matings, compared to the males of low genetic quality, thus in this way females can prevent deleterious alleles to be transmitted to the next generation. We tested this hypothesis through set up of two experimental groups from same genetic pool, where in one group genetic quality was manipulated by ionizing radiation. Within each group opportunity for choosing mates was imposed: males and females had no choice or had multiple choice. Mutational load was measured through the variability of different fitness components: fecundity and egg-to-adult viability. Our results indicate that sexual selection can reduce mutational load, only for fecundity. Group with the presence of female choice exhibited higher fecundity than group in which sexual selection was experimentally eliminated, but only in 'irradiated' group. There was no overall difference in egg-to-adult viability between different sexual selection regimes in any of the group. It should be considered that sexual selection can cause sexual conflict, and potential opposite effects of sexual selection and sexual conflict on fitness. Genetic structure of populations, in terms of the level of mutational load, is an important factor which can determinate the role of sexual selection.Prema teorijskim očekivanjima seksualna selekcija može smanjivati mutaciona opterećenja u populacijama, preko uspešnosti mužjaka u parenju. Mužjaci koji su boljeg genetičkog kvaliteta bi trebalo da budu uspešniji u parenju od mužjaka lošijeg genetičkog kvaliteta. Na taj način bi ženke mogle da smanje prenošenje štetnih alela u sledeću generaciju. Ova hipoteza je testirana uspostavljanjem dve eksperimentalne grupe od istog genetičkog pula jedinki, pri čemu su u jednoj mutacije indukovane jonizujućim zračenjem. Unutar svake grupe je nametnuta mogućnost izbora u parenju: mužjaci i ženke nisu imali izbor, ili su imali višestruki izbor. Mutaciona opterećenja su merena preko dve komponente adaptivne vrednosti: fekunditeta i preživljavanja od stadijuma jaja do adulta. Dobijeni rezultati ukazuju da seksualna selekcija može imati uticaj na smanjivanje mutacionih opterećenja ali samo za fekunditet. U prisustvu seksualne selekcije su vrednosti fekunditeta bile veće u odnosu na vrednosti koje su dobijene kad je seksualna selekcija eksperimentalno uklonjena, ali samo u okviru 'ozračene' grupe. Razlike u preživljavanju od jaja do adulta između različitih režima seksualne selekcije nisu dobijene ni u jednoj od grupa. Kako seksualna selekcija često uzrokuje i seksualni konflikt, treba uzeti u obzir njihovo potencijalno delovanje u suprotnim pravcima na adaptivnu vrednost. Genetička struktura populacija, koja podrazumeva i veličinu mutacionih opterećenja, može biti važan faktor od koga će i zavisiti delovanje seksualne selekcija na adaptivnu vrednost.Projekat ministarstva br. 17301

Nucleotide diversity of Cyt b gene in drosophila subobscura Collin

Mitochondrial DNA variability of Drosophila subobscura Collin from Southeastern Serbia was studied with respect to Restriction Site Analysis (RSA) of complete mitochondrial genome and the nucleotide sequence of Cytochrome b (Cyt b) gene. The aim was to shed more light on the evolutionary forces that shape mtDNA variation of this species. Samples were collected from two sites in the foothills of the Balkan Mountains. No genetic differentiation was found between groups and most of the variation was observed within them. Restriction analysis revealed two main haplotypes and several rare ones. The sequencing of Cyt b gene showed larger number of haplotypes, among which, one is being the most common. The majority of singletons differed from the most frequent haplotype by one nucleotide change. Although some of the observed nucleotide differences may affect their host’s fitness, the observed pattern of variation is consistent with the seasonal fluctuations in population size

Mating behavior as an indicator of quality of Drosophila subobscura males?

According to current theoretical predictions, any deleterious mutations that reduce nonsexual fitness may have a negative influence on mating success. This means that sexual selection may remove deleterious mutations from the populations. Males of good genetic quality should be more successful in mating, compared to the males of lower genetic quality. As mating success is a condition dependent trait, large fractions of the genome may be a target of sexual selection and many behavioral traits are likely to be condition dependent. We manipulated the genetic quality of Drosophila subobscura males by inducing mutations with ionizing radiation and observed the effects of the obtained heterozygous mutations on male mating behavior: courtship occurrence, courtship latency, mating occurrence, latency to mating and duration of mating. We found possible effects of mutations. Females mated more frequently with male progeny of nonirradiated males and that these males courted females faster compared to the male progeny of irradiated males. Our findings indicate a possible important role of sexual selection in purging deleterious mutations

Highly Contiguous Assemblies of 101 Drosophilid Genomes

Over 100 years of studies in Drosophila melanogaster and related species in the genus Drosophila have facilitated key discoveries in genetics, genomics, and evolution. While high-quality genome assemblies exist for several species in this group, they only encompass a small fraction of the genus. Recent advances in long-read sequencing allow high-quality genome assemblies for tens or even hundreds of species to be efficiently generated. Here, we utilize Oxford Nanopore sequencing to build an open community resource of genome assemblies for 101 lines of 93 drosophilid species encompassing 14 species groups and 35 sub-groups. The genomes are highly contiguous and complete, with an average contig N50 of 10.5 Mb and greater than 97% BUSCO completeness in 97/101 assemblies. We show that Nanopore-based assemblies are highly accurate in coding regions, particularly with respect to coding insertions and deletions. These assemblies, along with a detailed laboratory protocol and assembly pipelines, are released as a public resource and will serve as a starting point for addressing broad questions of genetics, ecology, and evolution at the scale of hundreds of species

Highly contiguous assemblies of 101 drosophilid genomes

Over 100 years of studies in Drosophila melanogaster and related species in the genus Drosophila have facilitated key discoveries in genetics, genomics, and evolution. While high-quality genome assemblies exist for several species in this group, they only encompass a small fraction of the genus. Recent advances in long-read sequencing allow high-quality genome assemblies for tens or even hundreds of species to be efficiently generated. Here, we utilize Oxford Nanopore sequencing to build an open community resource of genome assemblies for 101 lines of 93 drosophilid species encompassing 14 species groups and 35 sub-groups. The genomes are highly contiguous and complete, with an average contig N50 of 10.5 Mb and greater than 97% BUSCO completeness in 97/101 assemblies. We show that Nanopore-based assemblies are highly accurate in coding regions, particularly with respect to coding insertions and deletions. These assemblies, along with a detailed laboratory protocol and assembly pipelines, are released as a public resource and will serve as a starting point for addressing broad questions of genetics, ecology, and evolution at the scale of hundreds of species