The Unit of Selection in DROSOPHILA MERCATORUM. II. Genetic Revolution and the Origin of Coadapted Genomes in Parthenogenetic Strains

Drosophila mercatorum is a sexual species that can reproduce parthenogenetically. Previous studies revealed that parthenogenetic strains had "coadapted genomes" with high fitness under parthenogenesis and total homozygosity due to nonadditive and nonmultiplicative fitness interactions between chromosomal segments scattered throughout the genome. To study the evolutionary origins of such coadapted genomes, females from sexual matings in nature were isolated as virgins and challenged to reproduce parthenogenetically. Fitness studies were performed on genomes derived from these sexual females and upon their successful parthenogenetic progeny. By straddling the reproductive transition from sex to parthenogenesis, these fitness studies demonstrated that coadapted genomes arise immediately, apparently due to an intense selective bottleneck accompanying the reproductive transition, and are not due to the slow accumulation of epistatic complexes via mutation after parthenogenesis has already been established. The reproductive transition may also serve as an experimental model of the "genetic revolution" theory of speciation because the transition involves (1) the ultimate founder effect (one genome), (2) maximal genetic drift and fixation, (3) a drastic change in genetic environment characterized by total homozygosity, and (4) an intense selective bottleneck that interacts with the change in genetic environment and the need to adapt to a laboratory environment and a novel system of reproduction. Thus, all the elements theorized to underlie genetic revolution are present, albeit in extreme form. This study indicates that genetic revolutions are real phenomena that can quickly alter morphology, development, life history parameters and behavior. Indeed, the alterations can be so drastic that a new "species" evolves, complete with pre- and post-mating isolating mechanisms. However, isozyme loci do not appear to be the target of this genetic revolution, but rather loci regulating fundamental developmental processes. However, isozyme loci may be useful in predicting the a priori chance of a successful revolution since they can indicate how the population structure of the parent population influences levels of individual heterozygosity, the prime source of the genetic variability in the founder population that must pass through the selective bottleneck.     

Selection for Mating Reluctance in Females of a Diploid Parthenogenetic Strain of DROSOPHILA MERCATORUM

A diploid parthenogenetic strain of Drosophila mercatorum was outcrossed to produce genetic variance among the impaternate female offspring. Selection experiments were carried out for reluctance of the parthenogenetic females to mate.After only two cycles of selection, a parthenogenetic strain which is significantly less receptive to males from three different bisexual strains was obtained. It was also found that there is some degree of sexual isolation among the three bisexual strains used. The results support the idea that selection can render a newly produced diploid parthenogenetic strain behaviorally different from its bisexual ancestor. This appears to provide a framework which can explain the natural coexistence of diploid bisexual and diploid parthenogenetic biotypes in some species of insects.     

The Parthenogenetic Capacities and Genetic Structures of Sympatric Populations of DROSOPHILA MERCATORUM and DROSOPHILA HYDEI

Drosophila mercatorum is a sexual species that can reproduce parthenogenetically in the laboratory. A previous study showed that a natural population of D. mercatorum inhabiting the Kamuela garbage dump on the Island of Hawaii could produce both viable parthenogenetic adults and self-sustaining parthenogenetic lines. The present study deals with a second screen for parthenogenesis and an isozyme survey performed on natural populations of D. mercatorum and D. hydei caught in patches of Opuntia tuna about 10 kilometers from Kamuela. Both cactus-patch species produced viable parthenogenetic adults, but only D. mercatorum produced parthenogenetic females themselves capable of parthenogenesis. Moreover, D. mercatorum produced several "hot" lines characterized by high parthenogenetic rates, while all lines of D. hydei had a homogenous low rate. The parthenogenetic capacity of the cactus-patch D. mercatorum was lower than that of the garbage-dump D. mercatorum. Moreover, both the cactus-patch D. mercatorum and D. hydei had lower levels of polymorphism (26% and 22%, respectively) then the garbagedump D. mercatorum (44%), and both cactus-patch populations had heterozygote deficiencies with respect to Hardy-Weinberg equilibrium, unlike the garbage-dump population. Consequently, these data do not support the idea that decreased levels of heterozygosity in a sexual population increase the chance that sexual females will produce totally homozygous, parthenogenetic progeny.     

The Population Genetics of Parthenogenetic Strains of DROSOPHILA MERCATORUM. II. the Capacity for Parthenogenesis in a Natural, Bisexual Population

Drosophila mercatorum is a bisexual species, but certain strains are capable of parthenogenetic reproduction in the laboratory. We investigated the parthenogenetic capacity of the virgin daughters of females captured from a natural, bisexual population in Hawaii. An isozyme survey indicated the natural population is polymorphic at about 50% of its loci, and its individuals heterozygous at 18% of their loci. The predominant mode of parthogenesis in D. mercatorum causes homozygosity for all loci in a single generation. Despite this radical change in genetic state, 23% of the virgin female lines produced adult parthenogenetic progeny, and 16% produced parthenogenetic progeny themselves capable of parthenogenetic reproduction. The parthenogenetic rats as measured by the number of parthenogenetic progeny themselves capable of parthenogenesis divided by the number of eggs laid is arougn 10(-5) for the virgin female lines. We argue that one of the major reasons for this low rate is that very few of the impaternate zygotes have a genotype that can survive and reproduce under the genetic conditions imposed by parthenogenetic reproduction. This intense selective bottleneck can be passed in a single generation if enough unfertilized eggs are laid, and once passed is accompanied by a large (perhaps a thousandfold) increase in the rate of parthenogenesis and by modifications in many phenotypic traits such as morphology and behavior.     

Genetic Recombination and Clonal Selection in DROSOPHILA MERCATORUM

Discrete and continuous generation unisexual populations were established from parthenogenetic D. mercatorum females heterozygous for five visible loci and one electrophoretic locus, which marked all five major chromosome arms. In the first impaternate generation of the discrete generation population, all thirty-two possible recombinant genotypes for the five visible markers were displayed. However, 99% of these individuals were homozygous for all gene markers due to the predominant mode of parthenogenesis called nuclear duplication. Many of these homozygous individuals gave rise to isogenic clones that were compelled to compete with each other in subsequent generations. A detailed analysis of the genetic response of this clonal population showed strong evidence for selection involving epistatic interactions between linked and unlinked loci throughout the genome. The unit of selection is described as being determined as early as the zygotic stage of development.     

The Molecular through Ecological Genetics of Abnormal Abdomen in DROSOPHILA MERCATORUM. I. Basic Genetics

The abnormal abdomen syndrome (aa) in Drosophila mercatorum is characterized by the persistence of juvenilized cuticle on the adult abdomen. The aa phenotype is shown to depend on at least two X-linked genetic elements that are about one map unit apart near the centromeric end of the X chromosome. These two genetic elements are necessary for aa expression; one behaves as a dominant element and the other as a recessive. Overlaying these genetic studies upon molecular work reported elsewhere, it is argued that the dominant element is the presence of a 5 kb insertion in a majority of the X-linked repeats coding for the 28S ribosomal RNA. The recessive element appears to be a locus controlling differential replication of noninserted over inserted 28S genes during polytenization. The aa syndrome requires both the presence of the inserted repeats and the failure to preferentially amplify noninserted repeats. Given the necessary X-linked elements for aa, a variety of modifiers are revealed. First, aa expression in males is Y-linked, apparently corresponding to a deletion of the 18S/28S rDNA gene cluster normally found on the Y. Moreover, all major autosomes can modify the penetrance of aa.     

Meiosis in Male DROSOPHILA MELANOGASTER I. Isolation and Characterization of Meiotic Mutants Affecting Second Chromosome Disjunction

Two second chromosome, EMS-induced, meiotic mutants which cause an increase in second chromosome nondisjunction are described. The first mutant is recessive and causes an increase in second chromosome nondisjunction in both males and females. It causes no increase in nondisjunction of the sex chromosomes in either sex, nor of the third chromosome in females. No haplo-4-progeny were recovered from either sex. Thus, it appears that this mutant, which is localized to the second chromosome, affects only second chromosome disjunction and acts in both sexes.-The other mutant affects chromosome disjunction in males and has no effect in females. Nondisjunction occurs at the first meiotic division. Sex chromosome disjunction in the presence of this mutant is similar to that of sc(4)sc(8), with an excess of X and nullo-XY sperm relative to Y and XY sperm. In some lines, there is an excess of nullo-2 sperm relative to diplo-2 sperm, which appears to be regulated, in part, by the Y chromosome. A normal Y chromosome causes an increase in nullo-2 sperm, where B(s)Y does not. There is also a high correlation between second and sex chromosome nondisjunction. Nearly half of the second chromosome exceptions are also nondisjunctional for the sex chromosomes. Among the double exceptions, there is an excess of XY nullo-2 and nullo-XY diplo-2 gametes. Meiotic drive, chromosome loss and nonhomologous pairing are considered as possible explanations for the double exceptions.     

The Alcohol Dehydrogenase Polymorphism in Populations of DROSOPHILA MELANOGASTER. I. Selection in Different Environments

The allozyme polymorphism at the alcohol dehydrogenase locus in Drosophila melanogaster was studied in order to obtain experimental evidence about the maintenance of this polymorphism. Populations started with different initial allele frequencies from homozygous F and S lines showed a convergence of frequencies on regular food at 25 degrees, leading to values equal to those in the base populations. These results were interpreted as due to some kind of balancing selection. In populations kept at 29.8 degrees, a lower equilibrium F frequency was attained. Addition of ethanol and some other alcohols to the food gave a rapid increase in F frequency, and high humidity decreased the F frequency slightly. Combination or alternation of ethanol and high humidity had variable effects in the populations tested. For a further analysis of the allele-frequency changes, estimates were obtained for egg-to-adult survival under different conditions and for adult survival on ethanol-supplemented food. On ethanol food (both at regular and high humidity), egg-to-adult survival of SS homozygotes was considerably lower than that of the FF and FS genotypes. Under regular conditions of food, temperature and humidity, a tendency to heterozygote superiority was observed, while at high humidity a relative high survival of SS was noticed in some tests. Adult survival of SS was lower than that of FF, but FS was generally intermediate, though the degree of dominance differed between populations. The results are consistent with the hypothesis of the occurrence of selection at the Adh locus.     

Meiosis in Male DROSOPHILA MELANOGASTER. II. Nonrandom Segregation of Compound-Second Chromosomes

The segregation of compound-second chromosomes in males from two different stocks has been examined. Segregation is random in males from the C(2L)RM4, dp; C(2R)RM4, px stock. Gametes containing only one of the two compound chromosomes comprise 50% of the gametes, and gametes containing either both elements or neither element make up the other 50% of the gametes.——In males from the C(2L)RM, b; C(2R)RM, cn stock, gametes containing either C(2L)RM, b or C(2R)RM, cn make up the majority of the gametes. Gametes containing both chromosomes or neither chromosome account for only 2-3% of the gametes. The nonrandom segregation is due to the C(2R)RM, cn chromosome.——Viability is reduced in flies carrying the C(2R)RM, cn chromosome. This includes larval lethality, delayed development and premature adult lethality. Cytologically, this chromosome contains a large duplication of 2L material, which includes material proximal to region 38 or 39. It is suggested that the viability and segregational properties associated with this chromosome are due to the duplicated 2L material.     

Virility Deficiency and the Sex-Ratio Trait in DROSOPHILA PSEUDOOBSCURA. I. Sperm Displacement and Sexual Selection

The Sex-Ratio (SR) phenomenon is associated with the X chromosome of many species of Drosophila. Males carrying SR transmit predominantly sperm bearing the X chromosome. SR, therefore, has a very significant advantage in segregation. This paper provides an experimental analysis of the role of virility selection in preventing the fixation of SR. SR males are found to suffer substantial virility reduction when mated with nonvirgin females but not with virgin females. The reduced virility arises because they are weaker in sperm displacement and are discriminated against by nonvirgin females. The virility deficiency of SR males is even more severe at low temperatures.     

The Effects of Disruptive and Stabilizing Selection on Body Size in DROSOPHILA MELANOGASTER. I. Mean Values and Variances

Disruptive and stabilizing selection were applied to thorax and wing length in Drosophila melanogaster. Disruptive selection with negative assortative mating (D(-)) practiced on thorax length caused a large increase of the phenotypic variance; practiced on wing length the increase was less striking. Disruptive selection with random mating (D(R)) caused in most lines only a temporary increase in phenotypic variance, but mean values increased considerably. Stabilizing selection (S) on thorax length or wing length did not decrease the phenotypic variance, but the mean value of the selected character declined.-The proportion of flies emerging decreased in all lines, while development time increased. Variance of development time increased in the D(-)-lines. In both D(-)-lines the frequency of flies with an abnormal number of scutellars was high (> 60% in one of the lines) and there was a temporary increase in abnormal segmentation of the abdomen.     

A Cold-Sensitive Zygotic Lethal Causing High Frequencies of Nondisjunction during Meiosis I in DROSOPHILA MELANOGASTER Females

The X-linked, cold-sensitive zygotic lethal, l(1)TW-6(cs), both in homozygous and heterozygous females, induces nondisjunction of all four chromosomes at Meiosis I at both 25 degrees and 17 degrees . Nondisjunction frequencies approaching 0.5 for the X and fourth chromosomes have been observed at 16 degrees -18 degrees . The disjunction of the X chromosomes in males is not affected. The mutant causes mitotic irregularities in zygotes at both 25 degrees and 17 degrees . Mortality of all zygotes produced by the crosses 6(cs)/6(cs)x6(cs)/B(s)Y and FM7/6(cs)x6(cs)/B(s)Y is respectively 86% and 67-74% at 25 degrees and 99.8-99.9% and 94% at 17 degrees . The mortality of 6(cs) hemizygotes derived from females carrying no doses of 6(cs) (C(1)DX,y f/yx6(cs)/B(s)Y) is 45-55% at 25 degrees and 98% at 17 degrees . The length of the temperature-sensitive period for 6(cs) homo- and hemizygotes is affected by the maternal dosage of 6(cs); the shortest TSP is for zero and the longest is for two maternal doses. Mortality takes place primarily during embryogenesis with some larval and little pupal mortality. Analysis of sectioned embryos indicates that the large array of different patterns of damage observed could have arisen from abnormal cleavage divisions and the incomplete population of the blastoderm with nuclei.     

The Molecular through Ecological Genetics of Abnormal Abdomen. II. Ribosomal DNA Polymorphism Is Associated with the Abnormal Abdomen Syndrome in DROSOPHILA MERCATORUM

Restriction endonuclease cleavage analyses of cloned and genomic DNA samples indicate that the structure of the DNA encoding the large cytoplasmic RNAs (rDNAs) is altered in Drosophila mercatorum lines which exhibit an abnormal abdomen (aa) phenotype. In a majority of the rDNA repeat units from aa flies, the 28S coding sequence is interrupted by a large [5-6 kilobase pairs (kbp)] insert. A subclone containing this inserted DNA (ins 3) hybridizes primarily to rDNA-containing sequences in in situ and genomic blot hybridization experiments. Additionally, genomic nitrocellulose blot hybridization analyses show that ins- containing rDNA repeat units are clustered in a spontaneously arising aa mutant. This rDNA alteration in D. mercatorum flies with the aa phenotype more closely resembles the bobbed (bb) defect of D. hydei than the bb defect of D. melanogaster, which involves alterations in rDNA copy number. By analogy with the other Drosophila systems, we propose that the altered D. mercatorum rDNA repeat units are defective in rRNA production at a critical stage. The lowered levels of rRNA ultimately would limit the concentration of ribosomes needed to produce large quantities of a protein (in these cases, juvenile hormone esterase) needed for normal development.     

The Molecular through Ecological Genetics of Abnormal Abdomen. III. Tissue-Specific Differential Replication of Ribosomal Genes Modulates the Abnormal Abdomen Phenotype in DROSOPHILA MERCATORUM

The abnormal abdomen (aa) syndrome in Drosophila mercatorum is controlled by two major X-linked genetic elements. We have previously shown that the major X-linked element of aa is associated with the presence of large inserts in the 28S gene of the ribosomal RNA (rDNA) genes. We show that, in polytene tissue of wild-type D. mercatorum, the uninterrupted rDNA repeats are overreplicated relative to interrupted repeats. Uninterrupted rDNA repeats are also overreplicated in polytene tissue of hybrid larval offspring from wild-type and aa parents. This overreplication of uninterrupted repeats is not observed in diploid tissues of wild-type hybrids (of wild-type and aa parents) and homozygous aa larvae or in polytene tissue of aa larvae. Furthermore, molecular analysis of an aa line that has reverted to the wild type indicates that the reversion phenomenon is associated with the ability to overreplicate uninterrupted rDNA repeats in polytene tissues. The patterns of differential replication of rDNA genes in wild-type hybrids and aa larvae of D. mercatorum offer a possible mechanism for the tissue-specific control of the aa phenotype and suggest that the molecular basis for the second X-linked genetic element of aa is involved in the control of differential replication in polytene tissues.