Optimizing methyl‐eugenol aromatherapy to maximize posttreatment effects to enhance mating competitiveness of male Bactrocera carambolae (Diptera: Tephritidae)

Methyl‐eugenol (ME) (1,2‐dimethoxy‐4‐(2‐propenyl)benzene), a natural phytochemical, did enhance male Bactrocera carambolae Drew & Hancock (Diptera: Tephritidae) mating competitiveness 3 d after ingestion. Enhanced male mating competitiveness can significantly increase the effectiveness of the sterile insect technique (SIT). ME application to mass reared sterile flies by feeding is infeasible. ME application by aromatherapy however, would be a very practical way of ME application in fly emergence and release facilities. This approach was shown to enhance mating competitiveness of B. carambolae 3 d posttreatment (DPT). Despite this added benefit, every additional day of delaying release will reduce sterile fly quality and will add cost to SIT application. The present study was planned to assess the effects of ME‐aromatherapy on male B. carambolae mating competitiveness 1DPT and 2DPT. ME aromatherapy 1DPT or 2DPT did enhance mating competitiveness of B. carambolae males whereas ME feeding 1DPT and 2DPT did not. Male mating competitiveness was enhanced by the ME aromatherapy irrespective if they received 1DPT, 2DPT or 3DPT. ME aromatherapy, being a viable approach for its application, did enhance mating competitiveness of male B. carambolae 1 d posttreatment as ME feeding did 3 d after ingestion.     

Comparison of methyl eugenol metabolites, mitochondrial COI, and rDNA sequences of Bactrocera philippinensis (Diptera: Tephritidae) with those of three other major pest species within the dorsalis complex

Males of the oriental fruit fly, Bactrocera dorsalis (Hendel) and some of its sibling species have strong affinity for methyl eugenol (ME). Methyl eugenol ingested by male flies is biotransformed in the crop to two ME metabolites that eventually accumulate in the rectal gland, which is known to serve as a reservoir for B. dorsalis sex pheromones. When fed with ME, males of laboratory and wild B. philippinensis Drew and Hancock selectively accumulated two metabolites, 2-allyl-4,5-dimethoxyphenol and (E)-coniferyl alcohol, in the rectal gland, as was seen for B. dorsalis sensu stricto, B. invadens Drew, Tsuruta and White, and B. papayae Drew and Hancock. Phylogenetic analysis of COI and rDNA sequence data of these four taxa also revealed a close relationship among B. philippinensis, B. dorsalis s.s., B. invadens, and B. papayae (all four are members of the dorsalis species complex). This result corroborates pheromone analysis. The usefulness of pheromonal analysis as a chemotaxonomy tool to complement molecular and other analysis in differentiation of closely related sibling species within the Bactrocera dorsalis complex, for which use of morphological characters had been inadequate, is highlighted.     

Development of a genetic sexing strain in <Emphasis Type="Italic">Bactrocera carambolae</Emphasis> (Diptera: Tephritidae) by introgression of sex sorting components from <Emphasis Type="Italic">B. dorsalis</Emphasis>, Salaya1 strain

Background

The carambola fruit fly, Bactrocera carambolae Drew & Hancock is a high profile key pest that is widely distributed in the southwestern ASEAN region. In addition, it has trans-continentally invaded Suriname, where it has been expanding east and southward since 1975. This fruit fly belongs to Bactrocera dorsalis species complex. The development and application of a genetic sexing strain (Salaya1) of B. dorsalis sensu stricto (s.s.) (Hendel) for the sterile insect technique (SIT) has improved the fruit fly control. However, matings between B. dorsalis s.s. and B. carambolae are incompatible, which hinder the application of the Salaya1 strain to control the carambola fruit fly. To solve this problem, we introduced genetic sexing components from the Salaya1 strain into the B. carambolae genome by interspecific hybridization.

Results

Morphological characteristics, mating competitiveness, male pheromone profiles, and genetic relationships revealed consistencies that helped to distinguish Salaya1 and B. carambolae strains. A Y-autosome translocation linking the dominant wild-type allele of white pupae gene and a free autosome carrying a recessive white pupae homologue from the Salaya1 strain were introgressed into the gene pool of B. carambolae. A panel of Y-pseudo-linked microsatellite loci of the Salaya1 strain served as markers for the introgression experiments. This resulted in a newly derived genetic sexing strain called Salaya5, with morphological characteristics corresponding to B. carambolae. The rectal gland pheromone profile of Salaya5 males also contained a distinctive component of B. carambolae. Microsatellite DNA analyses confirmed the close genetic relationships between the Salaya5 strain and wild B. carambolae populations. Further experiments showed that the sterile males of Salaya5 can compete with wild males for mating with wild females in field cage conditions.

Conclusions

Introgression of sex sorting components from the Salaya1 strain to a closely related B. carambolae strain generated a new genetic sexing strain, Salaya5. Morphology-based taxonomic characteristics, distinctive pheromone components, microsatellite DNA markers, genetic relationships, and mating competitiveness provided parental baseline data and validation tools for the new strain. The Salaya5 strain shows a close similarity with those features in the wild B. carambolae strain. In addition, mating competitiveness tests suggested that Salaya5 has a potential to be used in B. carambolae SIT programs based on male-only releases.

     

Ecological significance of male attractant in the defence and mating strategies of the fruit fly, Bactrocera papayae

After being acclimatized to feeding on fruit flies, the Asian house gecko, Hemidactylus frenatus Duméril & Bibron (Gekkonidae:Squamata), consumed fewer methyl eugenol (ME) fed male fruit flies, Bactrocera papayae Drew & Hancock (Tephritidae: Diptera) than when offered ME-deprived males. After one-day exposure to only ME-fed males, the geckos avoided feeding on female flies when in the presence of ME-fed males. When mechanically disturbed, the ME-fed males spontaneously ejaculated a rectal secretion which contains phenyl propanoids that deter the predator.The ME-fed males also competed significantly better than normal (ME-deprived) males for virgin females. Male B. papayae converts ME to three other phenyl propanoids which act in concert as a sex pheromone to attract females during courtship and as an allomone to the gecko.     

Emergence, Mating, and Postmating Behaviors of the Oriental Beetle (Coleoptera: Scarabaeidae)

In a previous field-trapping study of the oriental beetle, Exomala orientalis (Waterhouse), by using synthetic sex pheromone on golf course fairways, numerous males were observed and trapped during the hours of peak mating activity. However, very few beetles were observed in the same areas when synthetic pheromone was absent. To investigate the hypothesis that mating in nature occurs cryptically within vegetation at the soil surface, laboratory studies on female emergence and pheromone release, male emergence and mate-locating, and female and male mating behaviors were conducted. Mate acquisition and copulation occurred on the soil surface near the female emergence site, with both sexes engaging in pheromone-mediated behaviors after having emerged from the soil. A highly stereotyped female pheromone release, or calling, behavior was observed, consisting of insertion of the female's head into the soil and elevation of the tip of her abdomen into the air. Bioassays conducted in a wind tunnel that simulated a turf fairway environment showed that walking and flying were both important in the upwind response of males to females. Mating and copulation occurred without an obvious complex courtship, but observations of postmating behaviors suggested that mate guarding occurs.     

Reproductive interference between two serious pests,oriental fruit flies <Emphasis Type="Italic">Bactrocera carambolae</Emphasis> and <Emphasis Type="Italic">B. dorsalis</Emphasis> (Diptera: Tephritidae), with very wide but partially overlapping host ranges

Bactrocera carambolae and B. dorsalis (Diptera: Tephritidae) are extremely destructive pests of fruits and vegetables in the Asia-Pacific region. Earlier reports have described that B. carambolae and B. dorsalis, respectively, use mainly star fruit and mango, suggesting a certain level of host partitioning which can be ascribed neither to differences in larval food qualities nor host-specific parasitoid mortality. This study specifically examined reproductive interference (antagonistic sexual interaction) between B. carambolae and B. dorsalis as a potential factor strongly affecting their host partitioning. We observed mating behaviors, especially interspecific courtships and mating, by cohabiting the conspecific and heterospecific pairs together. Consequently, we quantified their effects on the reproductive success of females. Males of both species frequently courted their own females, but they also courted females of other species. Courtship refusal by females was not selective in males of either species. This incomplete discrimination of both sexes led to frequent occurrences of interspecific sexual interactions in both species, but only B. carambolae females showed reduced reproductive success. These results suggest that B. dorsalis, superior in reproductive interference, can occupy high-quality mango, whereas B. carambolae, inferior in reproductive interference, must use low-quality star fruit.     

Protandry in the parasitoidCardiochiles nigriceps,as related to its mating system

A laboratory study was conducted of the emergence times and mating success ofCardiochiles nigriceps Viereck (Hymenoptera: Braconidae), a larval parasitoid of the tobacco budworm,Heliothis virescens (F.) (Lepidoptera: Noctuidae), and in view of this information, the parasitoid's mating system was explored. Observations on laboratory populations ofC. nigriceps confirmed the occurrence of two types of protandry, i.e. seasonal and diurnal; males emerged 2 days before females in a generation, and emerged about 1 hr before females on a given day. An experiment on mating success showed that newly emerged females are not receptive to males that emerged 1 hr earlier, but that these males and females often mate successfully after 1 more hr. The experiment also showed that 1- to 5-day-old males are more successful than 1-hr-old males in mating with 0- to 1-hr-old females. Thus, it is argued that males emerging on a given day have a disadvantage in competition for mates with males that emerged days earlier, and that this disadvantage may serve as a selection pressure toward diurnal protandry. A monogamous mating system for females ofC. nigriceps is suggested because sexual selection would be expected to be strong in species exhibiting both seasonal and diurnal protandry. A possibility of a sibling mating system inC. nigriceps is questioned partly because newly emerged females are unreceptive to males that emerged 1 hr earlier and partly because this parasitoid is solitary in what is considered highly dispersed hosts in the field.     

Condition-Dependent Mating Success in Male Fruit Flies: Ingestion of a Pheromone Precursor Compensates for a Low-Quality Diet

Previous studies revealed that males of the oriental fruit fly, Bactrocera dorsalis, require protein in the adult diet to obtain matings and that ingestion of methyl eugenol, which acts as a pheromone precursor, increases male attractiveness and mating competitiveness. The goal of this study was to investigate the interaction between diet quality and methyl eugenol consumption in affecting the mating frequency of B. dorsalis males. In one set of experiments, mature males were deprived of protein for 1, 3, or 7 days and were either given or denied access to methyl eugenol (ME). These males competed against control males (continuously protein-fed, no feeding on ME) for copulations in field cages. Without ME, males held without protein for 3 or 7 days obtained significantly fewer matings than control males. With ME, however, males held for even 7 days without protein achieved higher mating success than control males. In a second set of experiments, mature males were held without protein for 7 days and then given a protein-rich diet for 1, 3, or 7 days before testing and were either given or denied access to ME. Without ME, males were competitively inferior to control males when tested 1 or 3 days after resumption of protein feeding and equivalent to control males only after 7 days of protein feeding. With ME, however, males obtained significantly more matings than control males when tested 3 or 7 days after resumed protein feeding and had similar mating success as control males after 1 day of access to the protein-rich diet. Results show that mating success in this species is condition-dependent, with both nutritional state and ME consumption influencing male mating success. Under the test conditions, feeding on ME counteracted a low quality diet and enhanced male mating success.     

Pupation site and emergence time influence the mating success of bagworm females,Oiketicus kirbyi

In commercial oil palm plantations in Costa Rica, we tested the hypotheses that pupation site and emergence time affect the mating success of protogynous female bagworms,Oiketicus kirbyi (Guilding) (Lepidoptera: Psychidae). Greater proportions of female than male pupae on upper leaves of oil palms and greater proportions of mated females in the upper rather than lower crown strata support the hypothesis that selection of pupation site by female larvae influences the mating success of adults. Increasing captures of males with increasing trap height further suggest that enhanced mating success of females in tree tops may be attributed either to most effective dissemination of sex pheromone on higher sites, or to males foraging predominantly in the upper strata of oil palms. As the majority of females pupated in the middle rather than upper crown of oil palms, selection of pupation site by females may be affected by additional as yet unknown factors. Emergence of females significntly preceded emergence of males. Increasing proportions of mated females throughout the emergence seasons probably resulted from an increased ‘availability’ of males. In tropical rainforests with local variations inO. kirbyi developmental time and stage, protogyny may represent an evolutionary strategy that furthers outbreeding.     

Biology and fertility life table of Bactrocera carambolae on grape and acerola

Bactrocera carambolae Drew & Hancock (Diptera: Tephritidae), native of Southeast Asia, is present in Brazil but restricted to Amapá, Pará and Roraima, where it has quarantine pest status. The possible dispersion to other fruit producing regions of Brazil could cause damages, including for exportation, due to quarantine restrictions imposed by fruit importing countries. The objective of this work was to describe the biological parameters and calculate the fertility life table of B. carambolae on grape (Vitis vinifera L.) and acerola (Malpighia emarginata DC.). The experiment was conducted in the laboratory under controlled conditions. The mean number of punctures and the mean number of eggs per female on grapes were 1.48 ± 0.05 and 9.87 ± 0.36, respectively. The mean number puparia per fruit was 0.1 ± 0.02 (grape) and 0.5 ± 0.10 (acerola). The pupal viability was 82.4% (grape) and 70.6% (acerola). The mean time ± SE of one generation, in days, was 25.8 ± 1.10 (grape) and 19.7 ± 0.21 (acerola). The mean fecundity was 1663.8 ± 501.01 (grape) and 206.9 ± 26.21 (acerola) with eggs viability of 5.6% (grape) and 12.5% (acerola). The mean longevity, in days, was 77.3 ± 12.13 on grape and 82.4 ± 4.24 on acerola. The study found that B. carambolae completes its biological cycle on grape and on acerola leaving offspring.     

The effect of B chromosomes on mating success of the grasshopper Eyprepocnemis plorans

The mating ability of E. plorans was tested in laboratory conditions in six experimental units composed of ten males and fifteen females during 31 days. When significant differences were found (three from the six cages, and in totals) they involved a decrease of matings involving males with B chromosomes. The same tendency seems to exist in females, but to a lesser extent, so that a significant effect is only detected when the totals are considered. Accessory chromosomes also delay, in both sexes, the occurrence of the first mating. No mating preferences depending on the number of Bs were detected.     

Spatial Segregation of the Sexes and Nutrients affect Reproductive Success in a Dioecious Wind-pollinated Grass

In many dioecious plant species in which spatial distributions of males and females have been examined, the sexes are spatially segregated – usually along an environmental gradient. Unless pollen is uniformly distributed in a population, spatial segregation of the sexes should reduce the average mating success of individuals. In three Californian populations of Distichlis spicata – a wind-pollinated grass species that exhibits spatial segregation of the sexes – I examined patterns of pollen movement and the effects of pollen load and nutrient availability on seed set to determine whether spatial segregation of the sexes actually reduces mating success for both males and females. In two of the populations, pollen dispersal was restricted, and pollen augmentation consistently, significantly increased seed set. However, in the third population – which had the lowest seed set – I found that although there were some indications of pollen limitation, pollen dispersal was not restricted, and seed production was limited primarily by nutrient availability. These results imply that in some populations of D. spicata nutrient limitation on the production of seeds by females may be sufficiently strong that spatial segregation of the sexes causes a fairly low cost to reproductive success compared with a more random distribution of the sexes. However, in other populations, pollen does limit mating success, and the spatial segregation of males and females in these populations is reducing the fecundity of both males and females.     

Microsatellite markers for the pest fruit fly,Bactrocera papayae (Diptera: Tephritidae) and other Bactrocera species

The dorsalis complex contains some of the most economically important fruit fly pests of the Asia–Pacific regions, including Bactrocera dorsalis, Bactrocera papayae and Bactrocera carambolae. These species are morphologically indistinct and genetically very similar. We describe the development of 12 microsatellite markers isolated from a representative of the dorsalis complex, B. papayae. We show the potential utility of the B. papayae microsatellites and a set of microsatellites isolated from Bactrocera tryoni as population and species markers for the dorsalis complex.     

Suitability of stages of femaleEncarsia pergandiella [Hymenoptera: Aphelinidae] for development of conspecific male hyperprasites

Encarsia pergandiella Howard females develop as primary parasitoids of immature whiteflies, whereas ♀♀ develop as secondary parasitoids on ♂♂ of their own species or on other primary whitefly parasitoids. In this experiment, maleE. pergandiella development was examined in the laboratory at 24°C, using different stages of immatureE. pergandiella females [enclosed within the cuticle ofTrialeurodes vaporariorum (Westwood)] as hosts. Unmated adult femaleE. pergandiella were caged individually for 24 h on leaves of plants with either 5-day (early to late 3^rd instar larval), 7-day (late 3^rd instar to prepupal), or 9-day (pupal) hosts. In a control treatment, hosts were allowed to complete development without exposure to unmated adult females. Subsequent emergence of ♂♂ (hosts) and ♀♀ from all treatments was recorded daily. Pupal (9-day)E. pergandiella females were found to be the most suitable hosts for development of ♀♀. Nine- and 7-day hosts were attacked at a similar rate, as indicated by a similarly low proportion of host emergence, but ♀♀ emerged at a significantly higher rate in the 9-day treatment than in the 7-day treatment. Development time of ♀♀ was 15±1 days in the 9-day treatment, and 16±1 days in the 7-day treatment (p<0.005). The proportion of hosts emerging from the 5-day treatment was not significantly different from the control treatment. The data suggest that the colonizing ability ofE. pergandiella in greenhouses is likely to be limited by the delay of 7–9 days between oviposition of female eggs and the suitability of these ♂♂ for oviposition of male eggs. Since development time of the 2 sexes is similar this delay should cause a lack of synchrony in the emergence and mating of F₁ ♂♂ and ♀♀.      

Starvation and the Mating Success of Wild Male Mediterranean Fruit Flies (Diptera: Tephritidae)

Recent evidence suggests that the nutritional state of male Mediterranean fruit flies, Ceratitis capitata (Wied.) (medfly), is an important influence on various components of their reproductive biology, including mating success. The objective of the present study was to examine experimentally the effect of temporary starvation on the mating success of wild male C. capitata. Males were maintained on protein–sugar or sugar-only diets, and for each diet we compared the mating success of continuously fed males versus males starved for 18 or 24 h immediately before testing. In trials conducted on field-caged, host trees, males starved for 24 h obtained only about half as many matings as fed males for both diets. However, when the starvation period was 18 h, starved males reared on the protein–sugar diet mated significantly less frequently than fed males, whereas starved males reared on sugar mated as often as fed males. Measurements of male pheromone calling and female attraction revealed that reduced mating success likely reflected the decreased signaling activity of starved males.     

Sexual Selection of Zorion guttigerum Westwood (Coleoptera: Cerambycidae: Cerambycinae) in Relation to Body Size and Color

Zorion guttigerum is a flower-visiting longhorned beetle endemic to New Zealand. Sexual selection of this species in relation to the body size and color form of different sexes was investigated in the field. The population sex ratio, based on censuses of feeding and mating sites (flowers), is male-biased. Females are significantly larger than males. Both sexes have antennae of similar length but the antennal length relative to the elytral length is greater in males than in females, and the antennal length of males increases more with an increase in body size than that of females. Both sexes have dark blue (DB) and yellowish-brown (YB) individuals. Both pair-bonded and solitary males are similar in elytral and antennal length. In pair-bonded males, DB individuals are significantly more numerous than YB ones, but in solitary males, the number of both color forms is similar. Males tend to have territory protection behavior, fighting with and chasing away rival males from feeding and mating sites. Larger males usually win the fight but the size-dependent fighting advantage does not translate into mating success. Male color plays an important role in mating success, with DB males having a significantly better chance to mate than YB males. Furthermore, male body size and color also have interactions in mating success: males of DB color morph obtain a greater mating advantage according to body size. Pair-bonded females are significantly larger and have longer antennae than solitary females, suggesting that males prefer larger females for mating. In addition, females of DB color morph with longer antennae are also preferred by males for mating. The significance of these findings is discussed.     

Variations in carambola infestation rates byBactrocera carambolae drew and hancock (Diptera: Tephritidae) with fruit availability in a carambola orchard

The relationship between the infestation rate of carambola fruits byBactrocera carambolae Drew and Hancock was investigated in a carambola orchard. Phenology of the fruits was found to influence percentage infestation of fruits byB. carambolae. The proportion of unbagged or susceptible fruits infested varied with time of year and tended to decrease with the increase of unbagged fruits available on the tree. The number of ovipunctures per fruit varied with fruit size and was also found to be indicative of the number of adult insects (B. carambolae and parasitoids) that will emerge from each fruit. Higher number of susceptible fruits available on each tree also decreased both the number of ovipunctures per fruit and the number of eggs laid per fruit, which could possibly be due to the strategy of spreading the risk adopted by the femaleB. carambolae or a result of random selection with more hosts available. The main parasitoids recorded wereBiosteres vandenboschi (Fullaway) andB. arisanus (Sonan). The mean percentage of parasitism was 38.3% and it followed roughly that of infestation of fruits.     

Mating Behavior of Trissolcus basalis (Wollaston) (Hymenoptera: Scelionidae): Potential for Outbreeding in a Predominantly Inbreeding Species

The mating behavior of the quasi-gregarious egg parasitoid Trissolcus basalis (Wollaston) was investigated under field conditions. Trissolcus basalis has female-biased sex ratios and is a protandrous species, with males emerging 1–2 days before females. Males competed aggressively for control of the egg mass, with one male assuming dominance and control of the egg mass, although changes in dominance occurred at least once on each egg mass observed. Typical mating behavior involved the dominant male mating his sisters immediately upon their emergence from the egg mass. These behaviors are characteristic of an inbreeding species that manifests local mate competition. However, several aspects of the mating behavior of T. basalis are inconsistent with that of an inbreeding species. Over 18% of emerging females were not mated by the dominant male upon emergence, 13% of females were not observed to be mated at all and may have left their natal site as virgins, 25% of females were mated multiple times and sometimes by multiple males, females remained near the natal site for up to several hours after emergence before emigrating, and males dispersed away from the natal site during female emergence. Trissolcus basalis may be a predominantly inbreeding species but its emergence and mating behavior suggest that low-frequency outbreeding is also likely to occur.     

Piecing together an integrative taxonomic puzzle: microsatellite,wing shape and aedeagus length analyses of Bactrocera dorsalis s.l. (Diptera: Tephritidae) find no evidence of multiple lineages in a proposed contact zone along the Thai/Malay Peninsula

Bactrocera dorsalis (Hendel) and B. papayae Drew & Hancock represent a closely related sibling species pair for which the biological species limits are unclear; i.e. it is uncertain if they are truely two biological species, or one biological species which has been incorrectly split taxonomically. The geographical ranges of the two taxa are thought to abut or overlap on or around the Isthmus of Kra, a recognised biogeographic barrier located on the narrowest portion of the Thai Peninsula. We collected fresh material of B. dorsalis s.l. (i.e. B. dorsalis s.s.+ B. papayae) in a north–south transect down the Thai Peninsula, from areas regarded as being exclusively B. dorsalis s.s., across the Kra Isthmus, and into regions regarded as exclusively B. papayae. We carried out microsatellite analyses and took measurements of male genitalia and wing shape, both used previously to separate the taxa. No significant population structuring was found in the microsatellite analysis, consistent with one, predominantly panmictic population. Both morphological datasets showed consistent, clinal variation along the transect, without disjunction. No evidence supported historical vicariance driven by the Isthmus of Kra, and no dataset supported the current taxonomy of two species. Rather, within and across the area of range overlap or abutment between the two species, only continuous morphological and genetic variation was recorded. Recognition that morphological traits previously used to separate these taxa are continuous, and that there is no genetic evidence for population segregation in the region of suspected species overlap, is consistent with a growing body of literature that reports no evidence of biological differentiation between these taxa.     

Mating behavior and male mating success in wildAnastrepha Ludens (Diptera: Tephritidae) on a field-caged host tree

Mating behavior and factors affecting mating success of males were studied using wild Anastrepha ludens on a fieldcaged host tree. The most common courtship sequence had five components: (1) male calls from the underside of a leaf, (2) female arrives to the maleoccupied leaf, (3) male orients to female and stops calling, (4) one or both approach to a face-to-face position 1–3 cm apart, and (5) male mounts female after 1–2 s. Courtship behavior was almost identical to that of laboratoryculture flies observed previously under laboratory conditions. Most malefemale encounters occurred at a height of 1–2m, well inside the outer canopy of the tree. Differential mating success by males occurred. No male mated more than once per day, owing possibly to a very short sexual activity period. Factors favoring mating success of males were survival ability and tendency to join male aggregations and to fight other males. Thorax length and age (9–11 days difference) had no effects on male copulatory success. Overall win/loss percentage was not related to mating success because the males that were most successful at mating fought mostly among themselves, driving their win/loss percentage down. However, these successful males (at mating) won most of their fights against less successful males. Results confirmed a lek mating system: males aggregated, called, and defended territories; territories did not contain femalerequired resources; and females exercised mate choice, apparently through selection of sites within leks.