Radiation of the Drosophila nannoptera species group in Mexico.

The Drosophila nannoptera species group, a taxon of Mexican cactophilic flies, is an excellent model system to study the influence of abiotic and biotic factors on speciation, the genetic causes of ecological specialization and the evolution of unusual reproductive characters. However, the phylogenetic relationships in the nannoptera species group and its position within the virilis-repleta phylogeny have not been thoroughly investigated. Using a multilocus data set of gene coding regions of eight nuclear and three mitochondrial genes, we found that the four described nannoptera group species diverged rapidly, with very short internodes between divergence events. Phylogenetic analysis of repleta group lineages revealed that D. inca and D. canalinea are sister to all other repleta group species, whereas the annulimana species D. aracataca and D. pseudotalamancana are sister to the nannoptera and bromeliae species groups. Our divergence time estimates suggest that the nannoptera species group radiated following important geological events in Central America. Our results indicate that a single evolutionary transition to asymmetric genitalia and to unusual sperm storage may have occurred during evolution of the nannoptera group.

Egg size, embryonic development time and ovoviviparity in Drosophila species.

Lengths, widths and volumes of eggs from 11 species of Drosophila whose genomes have been fully sequenced exhibit significant variation that is not explained by their phylogenetic relationships. Furthermore, egg size differences are unrelated to embryonic development time in these species. In addition, two of the species, Drosophila sechellia and, to a lesser degree, D. yakuba, both ecological specialists, exhibit ovoviviparity, suggesting that female control over oviposition in these species differs from what is observed in D. melanogaster. The interspecific differences in these reproductive characters, coupled with the availability of whole genome sequences for each, provide an unprecedented opportunity to examine their evolution.

Post-zygotic isolation in cactophilic Drosophila: larval viability and adult life-history traits of D. mojavensis/D. arizonae hybrids.

Drosophila mojavensis and Drosophila arizonae are cactophilic flies that have been used extensively in speciation studies. Incomplete premating isolation, evidence of reinforcement, and a lack of recent introgression between these species point to a potentially important role for post-zygotic isolating barriers in this system. Other than hybrid male sterility, however, post-zygotic isolation between D. mojavensis and D. arizonae has received little attention. In this study, we examined viability and life-history traits of D. mojavensis/D. arizonae F(1) hybrids from sympatric crosses. Specifically, we reared hybrids and purebreds on the natural host cacti of each parental species and compared viability, development time, thorax length, and desiccation resistance between hybrids and purebreds. Interestingly, hybrid females from both crosses performed similarly or even better than purebred females. In contrast, hybrid sons of D. arizonae mothers, in addition to being sterile, had shorter average thorax length than males of both parental species, and hybrid males from both crosses had substantially lower desiccation resistance than D. mojavensis males. The probable cost to hybridization for D. mojavensis females resulting from reduced desiccation resistance of hybrid sons may have been an important selective factor in the history of reinforcement for crosses involving these females.

Mating preferences are not predictive of the direction of evolution in experimental populations of Drosophila.

The general applicability of two models used in predicting evolutionary directions from asymmetry in reproductive isolation was tested in the laboratory. In mate preference tests with strains of Drosophila melanogaster whose ancestral and derived relationships were known, no correspondence was found between sexual isolation and direction of evolution.

Drosophila males contribute to oogenesis in a multiple mating species.

Two species of Drosophila that differ in their ecology and mating systems have been compared with respect to male contribution to the somatic tissues and developing oocytes of females. In the species Drosophila mojavensis females remate daily, exhibit a copulatory plug, and have been shown to obtain a contribution from the male ejaculate. In contrast, Drosophila melanogaster males do not contribute to females. Female Drosophila melanogaster do not remate as frequently as Drosophila mojavensis females nor is a copulatory plug formed.

Conservation of the behavioral and transcriptional response to social experience among Drosophilids.

While social experience has been shown to significantly alter behaviors in a wide range of species, comparative studies that uniformly measure the impact of a single experience across multiple species have been lacking, limiting our understanding of how plastic traits evolve. To address this, we quantified variations in social feeding behaviors across 10 species of Drosophilids, tested the effect of altering rearing context on these behaviors (reared in groups or in isolation) and correlated observed behavioral shifts to accompanying transcriptional changes in the heads of these flies. We observed significant variability in the extent of aggressiveness, the utilization of social cues during food search, and social space preferences across species. The sensitivity of these behaviors to rearing experience also varied: socially naive flies were more aggressive than their socialized conspecifics in some species, and more reserved or identical in others. Despite these differences, the mechanism of socialization appeared to be conserved within the melanogaster subgroup as species could cross-socialize each other, and the transcriptional response to social exposure was significantly conserved. The expression levels of chemosensory-perception genes often varied between species and rearing conditions, supporting a growing body of evidence that behavioral evolution is driven by the differential regulation of this class of genes. The clear differences in behavioral responses to socialization observed in Drosophilids make this an ideal system for continued studies on the genetic basis and evolution of socialization and behavioral plasticity.

Phylogenetic relationships of leopard frogs (Rana pipiens complex) from an isolated coastal mountain range in southern Sonora, Mexico.

Mitochondrial DNA sequence data from the control region and 12S rRNA in leopard frogs from the Sierra El Aguaje of southern Sonora, Mexico, together with GenBank sequences, were used to infer taxonomic identity and provide phylogenetic hypotheses for relationships with other members of the Rana pipiens complex. We show that frogs from the Sierra El Aguaje belong to the Rana berlandieri subgroup, or Scurrilirana clade, of the R. pipiens group, and are most closely related to Rana magnaocularis from Nayarit, Mexico. We also provide further evidence that Rana magnaocularis and R. yavapaiensis are close relatives.

A genetic analysis of phototactic behavior in Drosophila melanogaster I. Selection in the presence of inversions.

The effectiveness of selection for positive and negative phototactic behavior in populations of Drosophila melanogaster heterozygous for various multiple inversions was compared using the method of realized heritability. Selection in the presence of FM6, SM1 or TM3 alone was as effective as in populations carrying no inversions. However, the presence of FM6 and TM3 together reduced the effectiveness of selection for photopositive behavior and FM6 and SM1 and TM3 restricted the response to selection for negative phototactic behavior. The results are discussed in terms of the organization of genes influencing phototactic behavior in this species.

Genetic analysis of phototactic behavior in Drosophila simulans.

Phototaxis mazes have been employed to select photopositive and photonegative strains of Drosophila simulans. The results suggest that phototactic behavior in D. simulans, as in other Drosophila species, is a polygenic trait. Hybridization using divergent strains revealed that the genes controlling negative phototactic behavior in D. simulans are autosomal, as opposed to D. melanogaster in which negative phototactic behavior is known to be very strongly sex-linked.

How Drosophila males make eggs: it is elemental.

Oogenesis in Drosophila requires a significant amount of phosphorus. Oocytes mature in follicles, each of which contains 15 highly polyploid, transcriptionally active chromosomes. We show that the demand for phosphorus is met in part from the male's ejaculate following mating. Females incorporate phosphorus-32 from radiolabelled males into their ovaries, specifically into their nucleic acids. Male-derived phosphorus is also present in significant amounts in mature oocytes. The mechanism by which phosphorus uptake from the female reproductive tract occurs must differ from that previously reported for radiolabelled carbon and hydrogen derived from ejaculatory proteins, as phosphorus uptake is observed in species not showing female incorporation of radiolabel derived from ejaculate proteins.

Reproductive behavior of Drosophila melanogaster and D. nigrospiracula in the field and in the laboratory.

The reproductive behaviors of two species of fruit fly, Drosophila melanogaster and D. nigrospiracula, were compared in field and laboratory populations. A number of differences were observed in the behavior of the two species in their natural habitats. D. melanogaster, which was observed on citrus, mates at its feeding site, whereas D. nigrospiracula, which is cactiphilic, mates on a non-resource-based male territory adjacent to its feeding site. In both species large male size is important for reproductive success. However, in D. melanogaster smaller males tended to be excluded from the breeding site and were therefore not among the pool of potential mates to which females were exposed. Sex ratios were biased toward females in both species, but the high frequency of female remating in D. nigrospiracula may have provided more mating opportunities for the males of this species. Field observations differed from laboratory observations, and I discuss the importance of these differences for understanding the evolution of Drosophila mating systems.

Fluctuating dermatoglyphic asymmetry and the genetics of liability to schizophrenia.

Schizophrenic subjects were compared to normal and psychiatric control subjects for degree of fluctuating asymmetry in two dermatoglyphic traits, a-b ridge count and fingertip pattern. The schizophrenic group exhibited significantly greater fluctuating asymmetry than either control group. Furthermore, indicators of disease severity such as early onset and declining course of illness correlated with degree of asymmetry. Both of these observations are expected if a disorder has a polygenic basis, since fluctuating asymmetry is a marker of polygenic inheritance.

Fluctuating dermatoglyphic asymmetry in psychotic twins.

Fluctuating asymmetry of bilateral morphological traits is the result of prenatal developmental instability and has been shown to be greater in organisms having more homozygous genotypes (aabb vs. AaBb, for example). This expected increase in fluctuating asymmetry has been found among individuals having a high degree of liability for schizophrenia, as this disorder appears to have a polygenic basis. We tested the additional prediction that the greater genetic liability for schizophrenia necessary for concordance between twins should be associated with greater fluctuating asymmetry in twin pairs in which both twins are mentally ill compared to twin pairs in which one individual is normal. Our analysis of asymmetry for finger ridge counts from fingerprints of concordant and discordant pairs of twins supports this prediction and provides additional indirect support for the roles of polygenic transmission and prenatal epigenetic vulnerability in schizophrenia.

Effects of starvation and desiccation on energy metabolism in desert and mesic Drosophila.

Energy availability can limit the ability of organisms to survive under stressful conditions. In Drosophila, laboratory experiments have revealed that energy storage patterns differ between populations selected for desiccation and starvation. This suggests that flies may use different sources of energy when exposed to these stresses, but the actual substrates used have not been examined. We measured lipid, carbohydrate, and protein content in 16 Drosophila species from arid and mesic habitats. In five species, we measured the rate at which each substrate was metabolized under starvation or desiccation stress. Rates of lipid and protein metabolism were similar during starvation and desiccation, but carbohydrate metabolism was several-fold higher during desiccation. Thus, total energy consumption was lower in starved flies than desiccated ones. Cactophilic Drosophila did not have greater initial amounts of reserves than mesic species, but may have lower metabolic rates that contribute to stress resistance.

Effects of age on water balance in Drosophila species.

Age-related declines in physiological performance have been documented in a wide variety of organisms. However, it is unknown whether related species age in a similar manner or whether physiological differences associated with aging differ widely among species. Previous work has shown that the desiccation resistance of Drosophila melanogaster decreases rapidly with age. Other Drosophila species may have delayed reproductive maturity or may inhabit arid habitats, so that age-related changes in water balance may be a significant physiological constraint on their ecological success. We may, therefore, predict that physiological systems involved in water balance will deteriorate with age at a reduced rate in these species. We quantified several components of water budgets for 14 species of Drosophila, including both desert and mesic representatives differing in their age of maturity. Desiccation resistance decreased with age in all but one species studied, primarily because of increased rates of water loss. However, there was no significant relationship between the rate of aging, as indicated by the rate of increase in water-loss rates as flies aged, and either habitat or age of maturity.

Evolutionary relationships of Drosophila mojavensis geographic host races and their sister species Drosophila arizonae.

The cactophilic Drosophila mojavensis species group living in the deserts and dry tropical forests of the southwestern United States and Mexico provides a valuable system for studies in diversification and speciation. Rigorous studies of the relationships between host races of D. mojavensis and the relationships among the members of the species group (D. mojavensis, Drosophila arizona, and Drosophila navojoa) are lacking. We used mitochondrial CO1 sequence data to address the phylogenetics and population genetics of this species group. In this study we have found that the sister species D. mojavensis and D. arizonae share no mitochondrial haplotypes and thus show no evidence for recent introgression. We estimate the divergence time between D. mojavensis and D. arizonae to be between 1.91 and 2.97 million years ago. D. arizonae shows little structure in our population genetic analyses but there is phylogenetic differentiation between southeastern and northern populations of D. arizonae. Drosophila mojavensis shows significant population and phylogenetic structure across the four geographic regions of its distribution. The mitochondrial data support an origin of D. mojavensis on the mainland with early differentiation into the populations now found in the Mojave Desert and the Mainland Sonoran Desert and later colonization of the Baja Peninsula, in contrast to previous models. Also, the sister clade to D. mojavensis/D. arizonae includes D. navojoa and Drosophila huaylasi. By defining the genetic relationships among these populations, we provide a foundation for more sophisticated hypothesis testing regarding the timing of early speciation events and host switches in this species group.

Microsatellite variation among diverging populations of Drosophila mojavensis.

Divergence and speciation may occur by various means, depending on the particular history, selective environments, and genetic composition of populations. In Drosophila mojavensis, a good model of incipient speciation, understanding the population genetic structure within this group facilitates our ability to understand the context in which reproductive isolation among populations is developing. Here we report the genetic structure and relationships of D. mojavensis populations at nuclear loci. We surveyed 29 populations throughout the distribution of D. mojavensis for four microsatellite loci to differentiation among populations of this species. These loci reveal four distinct geographical regions of D. mojavensis populations in the south-western United States and north-western Mexico--(i) Baja California peninsula (Baja), (ii) Sonora, Mexico-southern Arizona, United States (Sonora), (iii) Mojave Desert and Grand Canyon (Mojave), and (iv) Santa Catalina Island (Catalina). While all regions show strong isolation, Mojave and Catalina are highly diverged from other regions. Within any region, populations are largely homogenous over broad geographical distances. Based on the population structure, we find clear geographical barriers to gene flow appear to have a strong effect in isolating populations across regions for this species.

Genetic variation, population structure, and phylogenetic relationships of Triatoma rubida and T. recurva (Hemiptera: Reduviidae: Triatominae) from the Sonoran Desert, insect vectors of the Chagas' disease parasite Trypanosoma cruzi.

Nucleotide and amino acid sequence data from the mitochondrial cytochrome b (Cytb) and cytochrome c oxidase subunit I (COI) gene segments were used to gain insights into the population biology and phylogenetic relationships of two species of hematophagous kissing bugs (Hemiptera: Reduviidae: Triatominae) from the Sonoran Desert of northwestern Mexico and southern Arizona, USA, Triatoma rubida (Uhler, 1894) and T. recurva (Stål, 1868), both of which are vectors of the protozoan parasite Trypanosoma cruzi responsible for Chagas' disease. Analysis of molecular variance of gene sequences indicated significant structure among populations of both species from widely separated geographic localities. Phylogenetic analyses of gene and amino acid sequences employing both Bayesian and parsimony methods showed that T. recurva clustered within the phyllosoma complex of Triatoma species from central and southern Mexico with high statistical support, and that it was closely related to T. longipennis. Triatoma dimidiata also was shown to be closely related to the phyllosoma complex, as was T. sanguisuga which has historically been assigned to the lecticularia complex. Analyses of gene sequences were unable to confidently resolve relationships of T. rubida, although weak support for a T. nitida+T. rubida clade was seen under certain conditions. A provisional calibration of a mitochondrial DNA molecular clock for T. rubida, based on geological dates for the vicariant separation of the Baja California peninsula from mainland Mexico, suggested that pairwise sequence divergences for the Cytb and COI genes were 1.1-1.8% and 0.6-1.0% per million years, respectively. Two highly supported sympatric lineages of T. rubida uhleri from southern Arizona, which are hypothesized to have diverged approximately 550,000-900,000 years ago, were detected in the Cytb gene trees.

Nonantagonistic interactions between the sexes revealed by the ecological consequences of reproductive traits.

In addition to the obvious role reproductive traits play in mating-system evolution, reproductive characters can also have critical ecological or life history consequences. In this study we examine the ecological consequences of mating for female cactophilic Drosophila to test different hypotheses about the processes driving divergence in reproductive characters. Comparisons between intra- and interpopulation matings suggest that population differences in mating benefits, namely increased desiccation resistance in mated females, is not solely attributable to either a male or female-specific reproductive trait. Instead, the results indicate that increased desiccation resistance is a product of a male-female postmating-prezygotic interactions. The results underscore that postmating-prezygotic interactions can serve as an arena for the evolution of male characters that confer substantial benefits to females, not just costs arising from sexual conflict. Variation in the relative benefits conferred by mating between intra- and interpopulation matings also suggests that the relationship between speciation and divergence in reproductive characters via male-female interaction will be difficult to predict.

Gene expression patterns accompanying a dietary shift in Drosophila melanogaster.

The ability of many organisms to switch to new hosts can be critical to their survival in the wild. However, the genetic mechanisms underlying such shifts are poorly understood. In this study, we used complementary DNA (cDNA) microarrays to ask if changes in gene expression are observed in response to a dietary shift in Drosophila melanogaster, a dietary generalist. We found significant and repeatable differential expression in a number of genes related to metabolic function and stress, suggesting that a functional genomics approach will be useful in seeking loci involved in the ability of flies to utilize different resources.