Male mating success and survival in the field with respect to size and courtship song characters inDrosophila littoralis andD. montana (Diptera: Drosophilidae)

We investigated the importance of male song and morphological characters to the male mating success in a two-year field study in natural populations ofD. littoralis andD. montana. We compared the properties of mating flies with those of a random male sample taken at the same time and place. InD. littoralis the male's size had no effect on his mating success, while inD. montana small males had a mating advantage in the field during the first study year. Females preferred males with short sound pulses in both species. We also examined the relationship between male morphological and song characters and viability by collecting male flies in late summer and comparing the means of male characters to those of overwintered flies the next spring. InD. littoralis male size had no effect on overwinter survival. InD. montana large flies survived better than small flies. In both species the shifts in song characters during the winter dormancy were opposite to those caused by sexual selection. Our results, accordingly, imply a possible balance between the forces of sexual and natural selection, which act in opposing directions on attractive male traits.     

Interactions of the Males and Females of Three Sympatric Drosophila virilis-Group Species, D. montana, D. littoralis, and D. lummei, (Diptera: Drosophilidae) in Intra- and Interspecific Courtships in the Wild and in the Laboratory

Four species of the Drosophila virilis group, D. montana, D. littoralis, D. lummei, and D. ezoana, occur sympatrically in several locations in northern Europe. Courtship interactions between the flies of the three first-mentioned species were observed at malt baits in Kemi, northern Finland, to find out how the flies of different species recognize conspecific individuals and how interspecific courtships differ from intraspecific ones in the wild. Intraspecific courtships (including females of different reproductive stages) and interspecific courtships were also videotaped and analyzed in laboratory. In the wild the males courted both conspecific and allospecific females, even though the species varied in how much the males were attracted to females of different species. Interspecific courtships usually broke off when the male touched the female or when the male and/or the female vibrated his/her wings, producing acoustic cues. In the laboratory males courted conspecific females irrespective of the reproductive stage of the female, even though the courtships directed toward immature and fertilized females usually included only orienting and touching (no licking and singing). D. littoralis, and very rarely D. montana and D. lummei, males courted also allospecific females. In the few interspecific courtships between these three species, where the male proceeded to singing, females responded to male singing by vibrating their wings. This ended the courtship. It is suggested that both the chemical cues affecting female attractivity and the acoustic signals of males and females, which are produced by wing vibration, function in maintaining sexual isolation between these three species.     

Variation in male wing song characters inDrosophila plantibia (Hawaiian picture-wingedDrosophila group)

The males of most species of the HawaiianDrosophila planitibia group produce songs when vibrating their wings during courtship. In four of the most recently evolved species,D. differens, D. planitibia, D. heteroneura, andD. silvestris, these songs are simple in structure and possess a higher and more variable carrier frequency than the songs of the more ancestral species of the group. In the present paper, we studied the variation in wing song production and song characters in aD. planitibia population. Some males vibrated their wings at a very low amplitude or slow speed, producing no detectable sound. Other males produced sound bursts varying in carrier frequency and burst length. The carrier frequency of the song depended mainly on the wing posture of the male during wing vibrations and was consistent for individual males. Variation between males of different isofemale progenies was not significant in any measured song trait. Songs of the males recorded in the present study differed, however, from songs of the males of a laboratory strain recorded earlier.     

Differences in male courtship song among the species of thequinaria group ofDrosophila

Courtship song is known to vary among several groups of closely related species ofDrosophila. The present study investigates differences in song among four members of thequinaria group of fungal-breedingDrosophila which are known to coexist in Britain and the near-continent. Results show that the species can be distinguished by several song parameters including interpulse interval.D. phalerata is very different from the other three species, while there is considerable overlap betweenD. kuntzei andD. limbata in several characters.D. transversa appears to be more similar to the latter species than it is toD. phalerata. The question of mate choice and the potential for reproductive character displacement are discussed.     

CRITERIA OF FEMALE MATE CHOICE IN DROSOPHILA LITTORALIS,D. MONTANA,AND D. EZOANA

We examined sexual selection by Drosophila littoralis, D. montana, and D. ezoana females on male courtship sounds to determine whether the females use absolute or relative criteria when choosing their mates. Behavior of the females was observed, when they were courted by a single male producing normal sounds, or by a single wing-manipulated male producing abnormal sounds; and when they were courted by one or both of these males in a choice situation. The females usually accepted short-winged (but not wingless) males producing abnormal sounds, if they had no alternatives. However, if they heard the sound produced by a normal male, they rejected the deficient male. Drosophila littoralis and D. ezoana females selected between two wing-manipulated males with different wing areas. Our results suggest that the females choose their mates on the basis of relative criteria if the signals emitted by the courting males are within the range of acceptable cues.     

Testing the fisherian mechanism: examining the genetic correlation between male song and female response in waxmoths

Models of indirect (genetic) benefits sexual selection predict linkage disequilibria between genes that influence male traits and female preferences, owing to either non-random mate choice or physical linkage. Such linkage disequilibria, a genetic correlation, can accelerate the evolution of male traits and female preferences to exaggerated levels. But relatively few empirical studies have measured the genetic correlation between male traits and female responses in natural populations, and the findings of those few are mixed: often, genetic correlations are not found. We tested the above prediction in an acoustic pyralid moth, Achroia grisella, in which males attract females with a rhythmic train of sound pulses, and females respond only to song that exceeds a minimum pulse rhythm. Both male song rhythm and female threshold response are repeatable and heritable characters. Because female choice in A. grisella is based largely on male song, and males do not appear to provide direct benefits at mating, genetic correlation between male song rhythm and female response is expected. We studied 2 A. grisella populations, bred them according to a full-sib/half-sib design, split the progeny among 4 different environmental conditions, and measured the male song/female response genetic correlation in each of the 8 resulting groups. While song rhythm and response threshold were generally heritable, we found no evidence of significant genetic correlation between these traits. We suggest that the complexity of the various male song characters, of female response to male song, and of correlations between male song characters and between aspects of female response have mitigated the evolution of strong genetic correlation between song and response. Thus, exaggerated levels of trait development may be tempered.     

Calling song and selective phonotaxis in the field crickets,Gryllus firmus andG. pennsylvanicus (Orthoptera: Gryllidae)

The field cricket species, Gryllus firmusand G. pennsylvanicus,occur in a mosaic hybrid zone that roughly parallels the eastern slope of the Appalachian mountains in the northeastern United States. It is important to know what role, if any, the calling song plays in mate choice in sympatric and allopatric populations. In this report, we present results on the variability of calling song properties along transects across this hybrid zone. We also present the results of experiments on phonotactic selectivity of females from an allopatric population of G. firmus.The male calling song of allopatric G. firmuswas significantly slower in temporal rhythm (i. e., chirp and pulse repetition rates) and lower in pitch (i.e., dominant frequency) than that of allopatric G. pennsylvanicus.Calling song properties of males recorded in the hybrid zone varied considerably in temporal and spectral properties. In two-stimulus (choice) phonotaxis experiments, allopatric females of G. firmuspreferred synthetic calling songs with conspecific pulse repetition rates over songs that had lower and higher pulse rates. This preference persisted even when the sound pressure levels of alternative stimuli were unequal. Therefore, allopatric females of G. firmuscan discriminate between conspecific and heterospecific calling songs. Whether or not this same selectivity is present in sympatric populations remains unclear. Investigations of phonotactic selectivity in other allopatric and sympatric populations of both species are currently under way.     

THE GENETIC BASIS OF EVOLUTION OF THE MALE COURTSHIP SOUNDS IN THE DROSOPHILA VIRILIS GROUP

When courting, males of the Drosophila virilis group vibrate their wings and emit species-specific courtship sounds consisting of trains of polycyclic sound pulses. To analyze the genetic basis of evolutionary changes in the sounds we made an F₁ diallel set of reciprocal crosses between the members of the virilis phylad of the group (two stocks of D. virilis and one of D. americana americana, D. a. texana, D. novamexicana, and D. lummei). We also crossed the D. virilis stocks with the members of the montana phylad of the same group (D. kanekoi, D. littoralis, D. borealis, D. flavomontana, D. lacicola, and D. montana) and made a backcross (D. virilis x D. littoralis) x D. virilis using a D. virilis marker stock (b; sv t tb gp; cd; pe). The sounds of the hybrids were analyzed using the following parameters: the length of a pulse train (PTL), the number of pulses in a train (PN), the interpulse interval (IPI), the length of a pulse (PL), the number of cycles in a pulse (CN), and the length of a cycle (CL). In the virilis phylad, the differences between species appeared to be determined mainly by autosomal genes in each sound trait. The heritabilities (narrow-/broad-sense) obtained from the diallel tables were the following: PTL 0.662/0.817, PN 0.651/0.841, IPI 0.193/0.546, PL 0.408/0.552, CN 0.425/0.719, and CL 0.361/0.764. The direction of dominance is for longer PTL, higher PN and CN, and shorter IPI and CL. PL shows ambidirectional dominance. In the sounds of the virilis phylad species, PTL and PL seem to be phenotypically the most important parameters, since their components (PN and IPI for PTL, CN and CL for PL) are negatively correlated. In crosses between D. virilis and D. littoralis or D. flavomontana reciprocal hybrids differed from each other in PTL, IPI, PL, and CN indicating X-chromosomal or cytoplasmic inheritance. In the backcrosses between D. virilis and D. littoralis the role of the X chromosome was ascertained to be decisive. We conclude that an X-chromosomal major change allowing variation in IPI has occurred during the separation of the two D. virilis group phylads, the long IPI allowing variation also in PL (and CN). The evolution of the sounds in the virilis phylad has probably gone towards longer and denser pulse trains, while in the montana phylad the sounds have evolved in different directions.     

Nucleotide and repeat length variation at the nonA gene of the Drosophila virilis group species and its effects on male courtship song

Genes found to affect male courtship song characters in Drosophila melanogaster are good candidates when tracing genes responsible for species-specific songs in other Drosophila species. It has previously been shown that Thr–Gly repeat length variation at the period gene affects song traits in D. melanogaster, which gives the repetitive regions a special interest. In this work, we have characterised the patterns of nucleotide variation for gene regions containing two Gly and one Gln–Ala repeat in another D. melanogaster song gene, no-on-transient A, in D. virilis group species. The levels of nucleotide variability in D. virilis nonA were similar to those found for other genes of the species, and the gene sequences showed no signs of deviation from neutrality. The Gly 2 repeat preceding the central domain of the gene exhibited length variation, which did not, however, correlate with song variation either within D. virilis or between the species of D. virilis group. The Gly 3 repeat located on the other side of the central domain showed amino acid divergence parallel to the consensus phylogeny of the D. virilis group species. The species of the virilis subgroup having Asn after the first three glycines in this repeat have simple songs with no species-specificity, while the species of the montana subgroup having two Gly or Asn–Ser in this site have unique courtship songs. Amino acid differences between the species in this repeat may, however, reflect species phylogeny rather than have an effect on song divergence per se.     

In Situ Hybridisation Analysis of the X-Linked Genes in the Species of the Virilis Group of Drosophila

When estimating the level of DNA sequence variation within and between populations or when planning QTL analysis, it is essential to know the location of the genes under study. In the present work, five X chromosomal genes, earlier localised in Drosophila virilis and D. littoralis, were mapped by in situ hybridisation on the larval polytene chromosomes of four other virilis group species, D. a. americana, D. flavomontana, D. lacicola and D. montana. Conjugation of X chromosomes of the most interesting species pairs was studied in interspecific hybrids. Three of the marker genes were used as RFLP markers to examine the occurrence of recombination in D. flavomontana and D. montana hybrid females. The gene arrangement of all species studied, appeared to be different at the proximal end of the X chromosome, which prevented normal conjugation along the most part of the X chromosome. The data illustrating the locations of five X chromosomal marker genes are presented for D. a. americana, D. flavomontana, D. lacicola and D. montana.     

Genetics of sexual isolation based on courtship song between two sympatric species: Drosophila ananassae and D. pallidosa

Sexual isolation has been considered one of the primary causes of speciation and its genetic study has the potential to reveal the genetics of speciation. In Drosophila, the importance of courtship songs in sexual isolation between closely related species has been well investigated, but studies analysing the genetic basis of the difference in the courtship songs associated with sexual isolation are less well documented. Drosophila ananassae and Drosophila pallidosa are useful for studies of sexual isolation, because of their sympatric distribution and absence of postmating isolation. Courtship songs are known to play a crucial role in sexual isolation between these two species, and the female discrimination behaviour against the courting male has been revealed to be controlled by a very narrow region on the second chromosome. In this study we investigated the genetic basis controlling the song differences associated with their sexual isolation, using intact and wingless males with chromosomes substituted between species. The results obtained from F₁ hybrid males between these species indicate the dominance of the song characters favoured by D. pallidosa females. In addition, the results obtained from backcross F₂ males indicate that chromosome 2 had a major effect on the control of the song characters associated with sexual isolation.     

Courtship signals and mate choice of the flies of inbred Drosophila montana strains

We studied genetic variation in fly mating signals and mate choice in crosses within and between inbred strains of Drosophila montana. Male songs and the cuticular hydrocarbons of both sexes as well as some of the flies’ behavioural traits differed significantly between strains. This did not, however, cause sexual isolation between strains. In fact, courtship was shorter if the female was courted by a male of a foreign strain than when courted by their own male. Heterosis was found for courtship duration and the carrier frequency of male song. Diallel analysis of male song revealed additive genetic variation in four out of the five traits studied. Two traits showed dominance variation and one of these, carrier frequency, expressed unidirectional dominance with alleles for higher carrier frequency being dominant. Direction of dominance in carrier frequency was the same as the direction of sexual selection exercised by D. montana females on this trait, which suggests that sexual selection could be a driving force in the evolution of song towards a higher carrier frequency.     

Mating behavior and the evolutionary significance of mate guarding in three species of crane flies (Diptera: Tipulidae)

Three species of crane flies-Dactylolabis montana, Limonia simulans,and Antocha saxicola-gather near streams to mate and oviposit. All species are polygamous and sex ratios at these sites are male-biased. After a short mating bout, males guard females by standing over them during oviposition. Sperm competition appears to be intense and to follow last-male advantage, based on the packing of sperm within the two elongate spermathecae. Males of A. saxicolasuccessfully defend against rivals over 85% of the time. In contrast, defending males of D. montanaand L. simulanslose the female over 65% of the time during interactions with rivals. Despite the high frequency of loss, defending males gain additional oviposition time by engaging rivals in combat while the female continues to oviposit. Thus, a guarding male does not have to retain the female for guarding to be adaptive. Legs and claws of all species are sexually dimorphic and play an important role in guarding and defending.     

Cytogenetic and phytochemical investigations in hybrids of MacaronesianDryopteris (Pteridophyta: Aspidiaceae)

The cytology and phloroglucinol composition of wild and synthesized hybrids of MacaronesianDryopteris species have been examined. Both cytological and chemical evidence suggest that the tetraploidD. guanchica has arisen from the diploid speciesD. aemula andD. maderensis by hybridization and chromosome doubling. However, the formation of the characteristic fully aromatic compounds inD. aemula appears to be suppressed in most of the hybrids involving this species, and inD. guanchica.     

Natural and experimental hybridization inArmeria (Plumbaginaceae):A. villosa subsp.carratracensis

An experimental crossing program was carried out in order to test the hypothesis thatArmeria villosa subsp.carratracensis, a serpentine endemic from Southern Spain, is of hybrid origin. Fruit-set and mainly pollen stainability data in artificial hybrids demonstrate that internal interspecific barriers are weak. Two generations of backcrossing with the two putative parents —A. colorata andA. villosa subsp.longiaristata— can restore pollen stainability to an average of 83.2 and 68.7, respectively. A morphometric analysis of 148 specimens belonging to parental species, different artificial hybrid generations and wild putative hybrid race shows that (1) morphological characters have a strong genetic basis and thus are good markers in identifying hybrids and introgression inArmeria; (2) artificial backcrossing produces in morphometrical characters a variation which conforms in both trend and magnitude to what is commonly attributed in the wild to introgression; (3) the position of the putative hybrid race is intermediate between the two parental species used in the program. These results are in accordance with the hypothesis of hybrid origin ofA. villosa subsp.carratracensis. Natural and experimental hybridization inArmeria, I     

Androecial development and systematics inFlacourtiaceae s. l.

Androecial development of 13 species belonging to six tribes ofFlacourtiaceae has been investigated. While inScolopieae andFlacourtieae the stamens develop centrifugally, inErythrospermeae, Oncobeae andPangieae they are initiated in a centripetal sequence or a sequence that is neither distinctly centripetal nor centrifugal. The distribution of these developmental patterns coincides with the distribution of other characters (e.g. cyanogenic compounds, salicoid leaf teeth) and therefore supports a split of the family intoFlacourtiaceae s. str. (containing theScolopieae, Homalieae, Prockieae, Flacourtieae, Casearieae andBembicieae) andKiggelariaceae (withErythrospermeae, Oncobeae andPangieae) and is in accordance with results of recentrbcL studies.     

Morphological analysis ofAlouatta seniculus species group (Primates,Cebidae). A comparison with biochemical and karyological data

Fifteen cranial measurements were taken from wild caught specimens ofAlouatta seniculus seniculus, A. s. stramineus andA. s. macconnelli. A morphological analysis showed sex dimorphism in these three groups. A multivariate analysis discriminated among these taxa; males being more clearly discriminated than females. Our data showed that these taxa can be separated on the basis of quantitative cranial traits, biogeographic distribution, karyological differences, and biochemical characters. We therefore propose a new taxonomic arrangement, changing their taxonomic status to the species level (Alouatta seniculus, A. macconnelli, andA. stramineus).     

One- and two-dimensional polyacrylamide gel analysis of the heat shock proteins of the virilis group of Drosophila

The heat shock proteins of the virilis group of Drosophila are analyzed by one- and two-dimensional polyacrylamide gel analysis. This group consists of the two closely related but distinct virilis and montana phylads. The analysis reveals that some of the heat shock proteins are highly conserved among the two phylads while others are not. The 83-, 72-, and 69-kdalton proteins comigrate in all species examined. There is, however, a noticable trend toward greater molecular weight variability in the smaller heat shock proteins. In general, the heat shock protein patterns within each phylad follow the proposed phylogenetic relationships with some exceptions. D. ezoana and D. littoralis, both members of the montana phylad, exhibit heat shock protein patterns more similar to those of the virilis phylad. The data also demonstrate that the montana phylad has almost two times the heat shock allele members that the virilis phylad has. It is also shown that F₁ and F₂ hybrid flies of crosses between Drosophila species having different patterns of heat shock proteins show Mendelian segregation of alleles. After several generations of inbred growth, however, the pattern of heat shock protein synthesis in reciprocal hybrids each resembles that of the paternal parent. The implications of these findings are discussed.This research was supported in part by Damon Runyon-Walter Winchell Grant DRG-233F to R.M.S. and NIH Grant GM 27611 to R.V.S. R.V.S. is the recipient of an NIH Research Career Development Award.     

Sex determination in<Emphasis Type="Italic"> Drosophila melanogaster</Emphasis> and<Emphasis Type="Italic"> Musca domestica</Emphasis> converges at the level of the terminal regulator<Emphasis Type="Italic"> doublesex</Emphasis>

Sex-determining cascades are supposed to have evolved in a retrograde manner from bottom to top. Wilkins 1995 hypothesis finds support from our comparative studies in Drosophila melanogaster and Musca domestica, two dipteran species that separated some 120 million years ago. The sex-determining cascades in these flies differ at the level of the primary sex-determining signal and their targets, Sxl in Drosophila and F in Musca. Here we present evidence that they converge at the level of the terminal regulator, doublesex (dsx), which conveys the selected sexual fate to the differentiation genes. The dsx homologue in Musca, Md-dsx, encodes male-specific (MdDSX^M) and female-specific (MdDSX^F) protein variants which correspond in structure to those in Drosophila. Sex-specific regulation of Md-dsx is controlled by the switch gene F via a splicing mechanism that is similar but in some relevant aspects different from that in Drosophila. MdDSX^F expression can activate the vitellogenin genes in Drosophila and Musca males, and MdDSX^M expression in Drosophila females can cause male-like pigmentation of posterior tergites, suggesting that these Musca dsx variants are conserved not only in structure but also in function. Furthermore, downregulation of Md-dsx activity in Musca by injecting dsRNA into embryos leads to intersexual differentiation of the gonads. These results strongly support a role of Md-dsx as the final regulatory gene in the sex-determining hierarchy of the housefly.Edited by D. Tautz     

Prevalence of tick-borne pathogens in ixodid ticks (Acari: Ixodidae) collected from European wild boar (<Emphasis Type="Italic">Sus scrofa</Emphasis>) and Iberian red deer (<Emphasis Type="Italic">Cervus elaphus hispanicus</Emphasis>) in central Spain

Emerging tick-borne diseases of humans and animals have occurred frequently during the past 30 years. These disease outbreaks appear to result from changes in the distribution of tick and vertebrate hosts, and the introduction of humans and domestic animals into tick–pathogen–wildlife cycles. Use of molecular technologies now available for identification of pathogens in ticks can provide valuable information that allows for risk analysis of emerging tick-borne diseases. In this study, the prevalence of selected pathogens in ticks collected in six locations in central Spain from the major wild ungulate species, European wild boar (Sus scrofa) and Iberian red deer (Cervus elaphus hispanicus), was determined by PCR. Tick species collected included Ixodes ricinus, Dermacentor marginatus, Rhipicephalus bursa and Hyalomma m. marginatum. Pathogens identified in ticks included piroplasmids, Anaplasma spp., Ehrlichia spp. and Rickettsia spp. Piroplasmids were identified in all tick species except I. ricinus. Ehrlichia spp. were detected in all tick species and collection locations, while Rickettsia spp., which proved to be R. slovaca and a recently identified Rickettsia sp. DnS28, were identified only in D. marginatus. A. marginale and A. phagocytophilum were detected in D. marginatus, R. bursa and Hy. m. marginatum. Concurrent infections of these pathogens were frequently observed in ticks. Notably, A. phagocytophilum, which is infective for a broad host range that includes humans and domestic and wild animals, was identified in ticks from all collection locations. The variety of ticks and tick-borne pathogens demonstrated in this study suggests a risk in central Spain for the emergence of tick-borne diseases in humans and domestic animals.