Premating isolation is determined by larval rearing substrates in cactophilic Drosophila mojavensis. IX. Host plant and population specific epicuticular hydrocarbon expression influences mate choice and sexual selection.

Sexual signals in cactophilic Drosophila mojavensis include cuticular hydrocarbons (CHCs), contact pheromones that mediate female discrimination of males during courtship. CHCs, along with male courtship songs, cause premating isolation between diverged populations, and are influenced by genotype × environment interactions caused by different host cacti. CHC profiles of mated and unmated adult flies from a Baja California and a mainland Mexico population of D. mojavensis reared on two host cacti were assayed to test the hypothesis that male CHCs mediate within-population female discrimination of males. In multiple choice courtship trials, mated and unmated males differed in CHC profiles, indicating that females prefer males with particular blends of CHCs. Mated and unmated females significantly differed in CHC profiles as well. Adults in the choice trials had CHC profiles that were significantly different from those in pair-mated adults from no-choice trials revealing an influence of sexual selection. Females preferred different male CHC blends in each population, but the influence of host cactus on CHC variation was significant only in the mainland population indicating population-specific plasticity in CHCs. Different groups of CHCs mediated female choice-based sexual selection in each population suggesting that geographical and ecological divergence has the potential to promote divergence in mate communication systems.

Mate choice opportunity leads to shorter offspring development time in a desert insect.

We describe indirect genetic benefits of mate choice in two allopatric populations of cactophilic Drosophila mojavensis. By manipulating mate choice opportunity, we show that greater mate choice among sexually mature adults leads to shorter offspring egg-to-adult development times; the extent of this reduction was influenced by population origin and by host plant environment. We performed multiple-choice mating trials with individually marked flies to investigate whether differential male mating success was a consequence of female choice, male interaction, or both. We demonstrate that male copulation frequency was not random and instead, was determined by female choice. Virgin females in these trials were no less discriminating than females that had been previously exposed to males. These results suggest that there are indirect benefits of female mate choice that are population and environment specific, consistent with the hypothesis of ecologically influenced 'good genes' sexual selection.

Premating isolation is determined by larval rearing substrates in cactophilic Drosophila mojavensis. VIII. Mating success mediated by epicuticular hydrocarbons within and between isolated populations.

We tested the hypothesis that intrademic sexual selection has caused sexual isolation between populations of geographically isolated populations of cactophilic Drosophila mojavensis, and was mediated by epicuticular hydrocarbons (EHCs), contact pheromones in this system. Sexual selection and sexual isolation were estimated using a Baja California and mainland population by comparing the number of mated and unmated males and females in each of four pairwise population mating trials. EHC profiles were significantly different in mated and unmated males in the interdemic (Bajafemale symbol x Mainlandmale symbol and Mainlandfemale symbol x Bajamale symbol), but not the intrademic mating trials. A small number of EHCs was identified that best discriminated among mated and unmated males, mostly alkadienes with 34 and 37 carbons. Females showed population-specific preferences for male EHC profiles. However, EHC profiles between mated and unmated males in the intrademic mating trials were not significantly different, consistent with undetectable sexual selection estimated directly from numbers of copulating pairs vs. unmated adults. Thus, sexual isolation among populations was much stronger than sexual selection within these populations of D. mojavensis.

Hierarchical structure in the Drosophila mojavensis cluster (Diptera: Drosophilidae).

Sequences from three gene regions from the nuclear and mitochondrial genomes were used to examine the extent and nature of hierarchical structure in the Drosophilamojavensis cluster (Drosophila arizonae, D. mojavensis and D. navojoa) of the D. repleta species group. To determine the genetic divergence of these three species, sequence data were analyzed using maximum parsimony and population aggregation analysis. Individual and combined gene genealogies indicate that D. arizonae and D. mojavensis are neither diagnosable nor monophyletic with respect to one another. Although D. navojoa has differentiated from D. arizonae and D. mojavensis, as diagnosed by nuclear gene sequences, it may have undergone a reticulation event with D. arizonae. Our results suggest that either these taxa are still undergoing differentiation at the molecular level or have experienced gene flow in the recent past.

Polytene chromosomes as indicators of phylogeny in several species groups of Drosophila.

BACKGROUND: Polytene chromosome banding patterns have long been used by Drosophila evolutionists to infer degree of relatedness among taxa. Recently, nucleotide sequences have preempted this traditional method. We place the classical Drosophila evolutionary biology tools of polytene chromosome inversion analysis in a phylogenetic context and assess their utility in comparison to nucleotide sequences. RESULTS: A simultaneous analysis framework was used to examine the congruence of the chromosomal inversion data with more recent DNA sequence data in four Drosophila species groups - the melanogaster, virilis, repleta, and picture wing. Inversions and nucleotides were highly congruent with one another based on incongruence length difference and partitioned Bremer support values. Inversion phylogenies were less resolved because of fewer numbers of characters. Partitioned Bremer supports, corrected for the number of characters in each matrix, were higher for inversion matrices. CONCLUSIONS: Polytene chromosome data are highly congruent with DNA sequence data and, when placed in a simultaneous analysis framework, are shown to be more information rich than nucleotide data.

Epicuticular hydrocarbon variation in Drosophila mojavensis cluster species.

Epicuticular hydrocarbon variation was investigated among the three species of the Drosophila mojavensis cluster (D. mojavensis, D. arizonae, and D. navojoa) within the large D. repleta group. Because these hydrocarbons serve as contact pheromones in adult D. mojavensis, the chemical characteristics and differences in hydrocarbon profiles in populations of these three sibling species were further investigated. Twenty-seven hydrocarbon components with chain lengths ranging from C28 to C40, including n-alkanes, methyl-branched alkanes, n-alkenes, methyl-branched alkenes, and alkadienes were observed. Hydrocarbon profiles among the three species reared on different cactus hosts were easily aligned with previously identified components in D. mojavensis. Male and female D. navojoa possessed a 31-methyldotricont-6-ene absent in both D. arizonae and D. mojavensis, while lacking the 8,24-tritricontadiene present in these two species. D. navojoa adults had far less 2-methyloctacosane than these sibling species, but the significance of this difference was obscured by the degree of variation among populations in amounts of this hydrocarbon. Mainland and Baja California populations of D. mojavensis were fixed for differences in the amounts 8,24-tritricontadiene, 9,25-pentatricontadiene, and 9,27-heptatricontadiene, consistent with all previous studies. Amounts of 18 of the 27 hydrocarbon components were greater in flies reared on Opuntia cactus. Canonical discriminant function analysis resolved all three species into distinct, nonoverlapping groups, suggesting that epicuticular hydrocarbon profiles are species-specific in the D. mojavensis cluster. Based on the amounts of interpopulation variation in hydrocarbon profiles in these three species, we hypothesize that epicuticular hydrocarbon differences may evolve early during the formation of new species.

Premating isolation is determined by larval-rearing substrates in cactophilic Drosophila mojavensis. V. Deep geographic variation in epicuticular hydrocarbons among isolated populations.

Adult epicuticular hydrocarbon variation of 14 geographically isolated populations of cactophilic Drosophila mojavensis was assessed to further investigate mechanisms of sexual isolation. Hydrocarbon transfer experiments demonstrated that these compounds are part of the mate recognition system in this species. Sixteen of the 23 epicuticular hydrocarbon components studied differed in amounts between males and females, and 13 differed in quantity between the geographic regions encompassing Baja California and mainland Mexico (Sonora and Sinaloa). Eight hydrocarbon components, seven of which differed in quantity between sexes, showed significant sex-by-region interactions, indicating region-specific sex reversals in hydrocarbon quantities. Such regional variation in epicuticular hydrocarbon profiles suggests that these hydrocarbon differences have also evolved in D. mojavensis since this species invaded mainland Sonora and Sinaloa from Baja California by switching host plants, in addition to a number of key genetic, behavioral, and life-history characters.

Phylogenetic analysis of the repleta species group of the genus Drosophila using multiple sources of characters.

The species in the repleta group of the genus Drosophila have been placed into five subgroups-the mulleri, hydei, mercatorum, repleta, and fasciola subgroups. Each subgroup has been further subdivided into complexes and clusters. Extensive morphological and cytological analyses of the members of this species group have formed the foundation for the proposed relationships among the members of the repleta species group. Fifty-four taxa, including 46 taxa belonging to the repleta species group, were sequenced for fragments of four genes-16S ribosomal DNA (16S), cytochrome oxidase II (COII), and nitrogen dehydrogenase 1 (ND1) of the mitochondrial genome and a region of the hunchback (hb) nuclear gene. We also generated a partial data set of elongation factor 1-alpha (Ef1alpha) sequences for a subset of taxa. Our analysis used both DNA characters and chromosomal inversion data. The phylogenetic hypothesis we obtained supports many of the traditionally accepted clades within the mulleri subgroup, but the monophyly of taxonomic groups outside of this subgroup appears not to be supported. Phylogenetic analysis revealed one well-supported, highly resolved clade that consists of closely related members of the mulleri and buzzatii complexes. The remaining taxa, a wide assortment of taxonomic groups, ranging from members of other species groups to members of several subgroups and members of three species complexes from the mulleri subgroup are found in poorly supported arrangements at the base of the tree.

Studies of linkage in populations. XIII. A unique cause of linkage disequilibrium in natural populations of Drosophila robusta.

Natural populations of Drosophila robusta are polymorphic for chromosomal gene arrangements in most of its range, the deciduous forests of North America east of the Rocky Mountains. Many of the gene arrangements are the result of paracentric inversions on both arms of the metacentric second chromosome. They are frequently in linkage disequilibrium, determined in many areas largely, or entirely, by differing frequencies of cis and trans forms of the double heterokaryotypes and their component linkage combinations. Presence and degree of linkage disequilibrium in these populations varies with locality and gender, with males from southern localities exhibiting the largest deviation from equilibrium. Analysis of the extensive karyotype data encompassing the entire species range collected over the past 50 years shows that natural selection is primarily responsible for maintaining these complex polymorphisms.

Premating isolation is determined by larval rearing substrates in cactophilic Drosophila mojavensis. IV. Correlated responses in behavioral isolation to artificial selection on a life-history trait.

Studies of behavioral isolation among geographically isolated populations of Drosophila mojavensis have provided an understanding of incipient speciation wherein phylogeny and ecology play a prominent role. Populations of D. mojavensis in mainland Mexico and southern Arizona exhibit low but significant premating isolation from Baja California populations in laboratory mate choice tests. These same populations have undergone considerable life-history evolution in response to use of different host plants, suggesting that behavioral isolation between populations is a pleiotropic consequence of adaptation to different environments, or Mayr's geographic speciation hypothesis. This hypothesis was tested using bidirectional artificial selection on egg-to-adult development time in replicate lines of a mainland and Baja population cultured on two host cacti for 13 generations. Response to selection was greatest in the slow lines cultured on one host, yet there was uneven response in some lines due to variation in cactus tissue quality. Realized heritabilities for development time ranged from 0.04 to 0.16, which is consistent with previous estimates from half-sib/full-sib analyses of genetic variation. In most lines that responded to selection, premating isolation decreased to near zero. Correlated responses in behavioral isolation suggest that adaptation to contrasting environments can cause secondary responses in mate recognition systems that can influence the formation of new species.

Fitness correlates of spur length and spur asymmetry in male wild turkeys.

Tarsal spurs play an important role in intrasexual competition for females among male wild turkeys (Meleagris gallopavo). Thus variation in spur development may have important fitness consequences. Fitness correlates of spur development were studied in a free-living population of wild turkeys and it was found that heavier males and males with longer beards had longer spurs. Males that had longer spurs spent more time on display areas during the breeding season and less time moving among these areas compared to males with shorter spurs, independently of their body mass. Otherwise ideally symmetrical spurs showed fluctuating asymmetry between left and right tarsi, the degree of absolute asymmetry decreased with spur length in adults, but not in subadults, and males that survived at least one winter had more symmetrical spurs compared to males that did not. We conclude that if the ability to produce symmetrical spurs has a genetic basis, then spur length and spur asymmetry could reliably indicate individual quality and that these traits are under directional selection for increased size and symmetry in wild turkeys.

Remating effects on the genetic structure of female life histories in populations of Drosophila mojavensis.

A quantitative genetic analysis of nine adult fitness components was performed in two populations of cactophilic Drosophila mojavensis under natural conditions of fermenting cactus and ethanol vapour. Female progeny from 18 sires and 36 dams were treated to a range of six exposure periods to males to assess effects of remating frequency on female fitness. Lifetime fecundity increased with increasing male exposure, but longevity showed an intermediate optimum with temporary exposure to males of 2-4 days. Narrow-sense heritabilities were significant for egg production traits while broad-sense heritabilities were significant for longevity-related traits. Positive genetic correlations between components of fitness were expressed among functionally related traits, e.g. longevity was positively correlated with lifetime fecundity, the number of clutches laid, clutch size, and the number of eggs laid per day. Negative genetic correlations were detected between early and late life fecundity suggesting genetic tradeoffs among components of adult fitness.

Sensitivity to larval density in populations of Drosophila mojavensis: Influences of host plant variation on components of fitness.

Chromosomally polymorphic populations of Drosophila mojavensis from Baja California feed and breed on agria cactus, Stenocereus gummosus; whereas, monomorphic Arizona populations are associated exclusively with organ pipe cactus, S. thurberi. The effects of this host plant shift in expanding the kinds of feeding and breeding sites were assessed by manipulating larval density and recording differences in egg to adult development time and viability, and adult thorax size in both populations on artificially rotted substrates of both cactus species. Older agria rots increased development time but had no effect on viability. Organ pipe rots were qualitatively poorer substrates than agria rots for both monomorphic and polymorphic populations of D. mojavensis, especially at higher larval densities causing longer egg to adult development times, lower viabilities, and smaller thorax sizes than agria.The Baja population expressed shorter development times, higher viabilities, and smaller thorax sizes than the Arizona population on both cactus substrates. No evidence for cactus host race formation was found. The Baja population was less sensitive to increasing larval densities for all fitness characters studied on both cactus substrates indicating greater developmental homeostasis than in the monomorphic Arizona population. These data support the hypothesized central-marginal population structure within this species coincident with the distribution of host plants and lend insight into the process of adaptive divergence at different life history stages caused by host plant shifts.