Diversity of Anastrepha spp. (Diptera: Tephritidae) and associated braconid parasitoids from native and exotic hosts in southeastern Bahia, Brazil

We documented fruit fly-host associations and infestation rates over 5 yr in the state of Bahia, Brazil, by systematically collecting native and introduced fruits in backyard and commercial orchards, experimental stations, and patches of native vegetation. Fruit were collected in multiple sites in the southern and southernmost regions of Bahia. A total of 942.22 kg from 27 fruit species in 15 plant families was collected throughout this study. Of these, 15 plant species from six families were infested by Anastrepha species. A total of 11,614 fruit flies was reared from the fruit (5,178 females and 6,436 males). No specimens of Ceratitis capitata (Wiedemann) were recovered. Eleven Anastrepha species were recovered from the collected fruit: Anastrepha antunesi Lima (0.04%), Anastrepha distincta Greene (0.1%), Anastrepha fraterculus (Wiedemann) (53.5%), Anastrepha leptozona Hendel (4.5%), Anastrepha manihoti Lima (0.1%), Anastrepha montei Lima (1.0%), Anastrepha obliqua (Macquart) (33.0%), Anastrepha pickeli Lima (2.0%), Anastrepha serpentina (Wiedemann) (1.0%), Anastrepha sororcula Zucchi (3.0%), and Anastrepha zenildae Zucchi (1.8%). We recovered 1,265 parasitoids (Hymenoptera: Braconidae) from Anastrepha pupae. Three species of braconids were found to parasitize larvae of nine Anastrepha species. The most common parasitoid species recovered was Doryctobracon areolatus (Szépligeti) (81.7%), followed by Utetes anastrephae (Viereck) (12.2%) and Asobara anastrephae (Muesebeck) (6.1%). We report A. fraterculus infesting Malay apple Syzygium malaccense (L.) Merr. & L. M. Perry and A. fraterculus, A. sororcula, and A. zenildae infesting araza Eugenia stipitata McVaugh for the first time in Brazil.     

Hosts and parasitoids of fruit flies (Diptera: Tephritoidea) in the State of Tocantins, Brazil

Fruit flies were obtained from 13 species of naturally infested fruits in the central region of Tocantins State, from January to October 2005. A total of 1,753 female flies were collected that belong to 11 species: Anastrepha coronilli Carrejo & González, A. fraterculus (Wied.), A. mucronota Stone, A. obliqua (Macquart), A. sororcula Zucchi, A. striata Schiner, A. turpiniae Stone, A. zenildae Zucchi, Anastrepha sp., Ceratitis capitata (Wied.) and Neosilba sp. Also six species of parasitoids were associated to Anastrepha larvae: Asobara anastrephae (Muesebeck), Doryctobracon areolatus (Szépligeti), Doryctobracon sp., Opius bellus Gahan, Opius sp. and Utetes anastrephae (Viereck).     

The Identity of Specimens of the Anastrepha fraterculus Complex (Diptera, Tephritidae) with Atypical Aculeus Tip

Several specimens collected in Paraguay along with Anastrepha fraterculus (sensu lato) have an aculeus tip similar to species from the fraterculus complex, but the teeth of the aculeus of these specimens are poorly defined. As Anastrepha species identification is based mostly on subtle differences in the aculeus tip, we studied these specimens with atypical aculeus tips (with poorly defined teeth) that slightly differs from the aculeus tip of species of the fraterculus complex (with well-developed blunt teeth), to determine if this is due to intraspecific variation or if it can characterize a full species. The Paraguayan specimens were separated in six groups under stereomicroscope according to variation in their aculeus tip. Specimens within each group were studied by means of morphometrics (traditional and geometric) and gene sequence analysis (COI and ITS1). Morphometric analyses were significant, but no clear groups were formed by the discriminant analyses of the aculeus and wing, and the COI and ITS1 sequence analysis clustered specimens with all six aculeus variations. Therefore, the subtle morphological differences observed in the aculeus tip of Paraguayan specimens are intraspecific variations and the Paraguayan specimens were more genetically closely related to Anastrepha sp. 3 from the fraterculus complex.     

Fruit flies (Diptera: Tephritidae) and their parasitoids (Hymenoptera: Braconidae) associated to host plants in the southern region of Bahia State

The association among Anastrepha species, braconid parasitoids and host fruits in southern Bahia is recorded. Doryctobracon areolatus (Szépligeti) was associated with A. serpentina (Wied.) in Pouteria caimito, A. bahiensis Lima in Helicostylis tomentosa, A. sororcula Zucchi in Eugenia uniflora, and A. obliqua (Macquart) in Spondias purpurea. Anatrepha obliqua was unique in fruits of Averrhoa carambola, but associated with D. areolatus, Asobara anastrephae (Muesebeck) and Utetes anastrephae (Viereck). In Achras sapota, A. serpentina was associated with A. anastrephae and D. areolatus, while in Psidium guajava, A. fraterculus (Wied.) and A. sororcula were associated with D. areolatus and U. anastrephae.     

Morphometric analysis of Mexican and South American populations of the Anastrepha fraterculus complex (Diptera: Tephritidae) and recognition of a distinct Mexican morphotype

Discriminant function and cluster analyses were performed on 19 morphometric variables of the aculeus, wing and mesonotum to determine whether populations of Anastrepha fraterculus (Wiedemann) from different parts of Mexico could be distinguished from populations from South America. Samples were collected from seven localities across Mexico, two from Brazil, and one each from Colombia and Argentina. Results showed there were statistically significant differences between Mexican and South American populations with respect to the aculeus (tip length, length of serrated section, mean number of teeth) and wing (width of S-band and connection between S- and V-bands). The degree of morphological variation observed among Mexican populations was extremely low, and as a consequence, the Mexican populations were identified as a single morphotype by discriminant analysis. The 'Andean morphotype'consisting of the Colombian population, and the 'Brazilian morphotype'consisting of the two Brazilian populations plus the single Argentinian population were also distinguished. It was concluded that the macro-geographical morphotypes from Mexico, Colombia, and Brazil plus Argentina correspond to three distinct taxonomic entities. Comparisons of results with those obtained from behavioural, karyotypic, isozyme and DNA studies suggest that sufficient evidence now exists to name a new Mexican species from within the A. fraterculus complex. This will be done in a separate publication. A provisional key to the morphotypes of A. fraterculus studied is provided.     

Faunistic analysis of fruit flies (Diptera: Tephritidae) in the Northern and Northwestern Regions of Rio de Janeiro State, Brazil

This study aimed to characterize the fruit fly populations in three municipalities of the Northern region and two municipalities of the Northwestern region of Rio de Janeiro State, Brazil, and to evaluate the similarity among their populations. A faunistic analysis was performed from the fruit fly specimens captured in plastic McPhail traps with an aqueous solution of hydrolyzed protein to 5% placed in orchards of guava (Psidium guajava L.) and/or other fruits during 26 months. The total of 3,952 females of 15 species of Anastrepha Schiner and 277 females of Ceratitis capitata (Wied.) was captured. The species richness differed among the municipalities, with the highest value in S?o Francisco do Itabapoana (S = 14), resulting on the highest Shannon-Wiener index (H = 1.27). The equitability was low in the five municipalities due to the dominance of one unique fruit fly species. The predominant species (more frequent, constant and dominant) were Anastrepha obliqua (Macquart) in Campos dos Goytacazes and S?o Francisco do Itabapoana, Anastrepha fraterculus (Wied.) in Cambuci and Itaocara, and Anastrepha sororcula Zucchi in S?o Jo?o da Barra. Fruit fly populations had low diversity index of Margalef (alpha = 0.58 to 1.82). Regarding to fruit fly species composition, the populations in S?o Jo?o da Barra and Cambuci were more similar between each other, composing a distinct group from the populations in Campos dos Goytacazes and Itaocara. These two groups differed quite a lot from the population in S?o Francisco do Itabapoana.     

Long aculeus and behavior of Anastrepha ludens render gibberellic acid ineffective as an agent to reduce 'ruby red' grapefruit susceptibility to the attack of this pestiferous fruit fly in commercial groves

Treating Mexican grapefruit with gibberellic acid (GA3) before color break, significantly delayed peel color change and increased peel puncture resistance, but it did not reduce grapefruit susceptibility to Mexican fruit fly, Anastrepha ludens (Loew) attack under natural conditions. Despite GA3 treatments, larval infestation levels increased with higher fruit fly populations, which also increased as the season progressed. Late in the season, infestation levels were even higher in GA3-treated fruit compared with untreated fruit, possibly because treated fruit were in better condition at that stage. Egg clutch size was significantly greater in very unripe, hard, GA3-treated fruit at the beginning of the harvest season and in December, compared with control fruit. Under laboratory conditions, egg injection into different regions of the fruit suggested that A. ludens eggs are intoxicated by peel oil content in the flavedo region. However, A. ludens' long aculeus allows females to oviposit eggs deeper into the peel (i.e., albedo), avoiding toxic essential oils in the flavedo. This makes A. ludens a particularly difficult species to control compared with other citrus-infesting species such as Anastrepha suspensa (Loew), Anastrepha fraterculus (Wiedemann), and Ceratitis capitata (Wiedemann) (fly species with significantly shorter aculei), which can be effectively managed with GA3 sprays. We discuss our findings in light of their practical implications and with respect to the oviposition behavior of various fruit flies attacking citrus.     

Nonhost status of Citrus sinensis cultivar valencia and C. paradisi cultivar ruby red to Mexican Anastrepha fraterculus (Diptera: Tephritidae)

Anastrepha fraterculus (Wiedemann) is recognized as a pest of citrus, apples, and blackberries in South America. In Mexico, it is mainly found in fruit of the family Myrtaceae and has never been reported infesting citrus. Here, we sought to determine whether females stemming from Mexican A. fraterculus populations (collected in the state of Veracruz) would lay eggs in 'Valencia' oranges and 'Ruby Red' grapefruit and, if so, whether larvae would hatch and develop. We worked under laboratory and seminatural conditions (i.e., gravid females released in fruit-bearing, bagged branches in a commercial citrus grove) and used Anastrepha ludens (Loew), a notorious pest of citrus, as a control species. Under laboratory conditions, A. ludens readily accepted both oranges and grapefruit as oviposition substrates, but A. fraterculus rarely oviposited in these fruit (but did so in guavas, a preferred host) and no larvae ever developed. Eggs were deposited in the toxic flavedo (A. fraterculus) and nontoxic albedo (A. ludens) regions. Field studies revealed that, as was the case in the laboratory, A. fraterculus rarely oviposited into oranges or grapefruit and that, when such was the case, either no larvae developed (oranges) or of the few (13) that developed and pupated (grapefruit), only two adults emerged that survived 1 and 3 d, respectively (5-17% of the time necessary to reach sexual maturity). In sharp contrast, grapefruit exposed to A. ludens yielded up to 937 pupae and adults survived for >6 mo. Therefore, the inability of Mexican A. fraterculus to successfully develop in citrus renders the status of Mexican A. fraterculus as a pest of citrus in Mexico as unsubstantiated.     

Diversity of flies (Diptera: Tephritidae and Lonchaeidae) in organic citrus orchards in the vale do Rio Caí, Rio Grande do Sul, southern Brazil

This study was carried out in the Counties of Montenegro and Pareci Novo located in the region of the Vale do Rio Cai, Rio Grande do Sul, Southern Brazil, aiming to determine the fruit fly species of Tephritidae and Lonchaeidae that occur in organic orchards of sweet orange [Citrus sinensis (L.) Osb.] cultivar Céu, and Murcott tangor (Citrus reticulata Blanco x C. sinensis), during the fruit ripening stages in 2003 and 2004. Eight McPhail traps baited with integral grape juice diluted to 25% were installed in four orchards, two in each citrus species. The traps were checked weekly, when the baits were changed, the flies separated and preserved in 70% ethyl alcohol. Fruits were also sampled from the orchards, placed in containers with damp soil and closed with a mesh. The tephritid flies represented 86.2% of all captured flies in the four orchards during both years. Five Tephritidae species were captured from traps: Anastrepha fraterculus (Wiedemann), Anastrepha grandis (Macquart), Anastrepha pseudoparallela (Loew), Anastrepha dissimilis Stone and Ceratitis capitata (Wiedemann). The captured species of Lonchaeidae were: Neosilba zadolicha McAlpine & Steyskal, Neosilba n.sp.3, Neosilba sp. and Lonchaea sp. Anastrepha fraterculus was found in 99% of the fruit samples, both in 'Céu' orange and 'Murcott' tangor, and Neosilba n.sp.3 were only obtained from 'Murcott' tangor fruits.     

Native and introduced host plants of Anastrepha fraterculus and Ceratitis capitata (Diptera: Tephritidae) in northwestern Argentina

Wild or commercially grown, native and exotic fruit were collected in 30 localities in the Tucumán province (NW Argentina) from January 1990 to December 1995 to determine their status as hosts of Anastrepha fraterculus (Wiedemann) and/or Ceratitis capitata (Wiedemann), the only two fruit fly species of economic and quarantine importance in Argentina. A total of 84,094 fruit (3,466.1 kg) representing 33 species (7 native and 26 exotic) in 15 plant families were sampled. We determined the following 17 host plant associations: Annona cherimola Miller (Annonaceae), Citrus paradisi Macfadyn (Rutaceae), Diospyros kaki L. (Ebenaceae), Eugenia uniflora L., Psidium guajava L., Myrcianthes pungens (Berg) Legrand (Myrtaceae), Ficus carica L. (Moraceae), Juglans australis Grisebach (Juglandaceae), Mangifera indica L. (Anacardiaceae), Eriobotrya japonica (Thunb.) Lindl., Prunus armeniaca L., P. domestica L., and P. persica (L.) Batsch (Rosaceae) were infested by both A. fraterculus and C. capitata. Citrus aurantium L., Citrus reticulata Blanco, Citrus sinensis (L.) Osbeck (Rutaceae), and Passiflora caerulea L. (Passifloraceae) were only infested by Ceratitis capitata. Out of a total of 99,627 adults that emerged from pupae, 69,180 (approximately 69.5%) were Anastrepha fraterculus, 30,138 (approximately 30.2%) were C. capitata, and 309 (approximately 0.3%) were an unidentified Anastrepha species. Anastrepha fraterculus predominated in native plant species while C. capitata did so in introduced species. Infestation rates (number of larvae/kg of fruit) varied sharply from year to year and between host plant species (overall there was a significant negative correlation between fruit size and infestation level). We provide information on fruiting phenology of all the reported hosts and discuss our findings in light of their practical (e.g., management of A. fraterculus and C. capitata in citrus groves) implications.     

Susceptibility of guava genotypes to natural infestation by Anastrepha spp. (Diptera: Tephritidae) in the municipality of Monte Alegre do Sul, state of São Paulo, Brazil

The infesting species and their infestation indices of fruit flies were determined for eleven guava genotypes (Psidium guajava L.). From March to April 2000, ten mature fruits of each genotype were harvested at weekly intervals from insecticide unsprayed trees located in the municipality of Monte Alegre do Sul, SP, Brazil. Fruits were brought to the laboratory, weighed and placed in individual plastic cups containing sand at the bottom to obtain the tephritid pupae. About 95% of guavas produced fruit fly puparia. Of the 682 Anastrepha females recovered, four species were identified: A. fraterculus (Wied.) (86.5%), A. obliqua (Macquart) (10.8%), A. bistrigata Bezzi (1.8%) and A. sororcula Zucchi (0.9%). Three species of parasitoids Braconidae (Opiinae) were recovered: Doryctobracon areolatus (Szépligeti), Doryctobracon brasiliensis (Szépligeti) and Utetes anastrephae (Viereck). The genotypes differ in level of infestation depend on the collecting time. The genotypes 'L2P4 Vermelha', 'Ruby Suppreme' and 'Webber Suppreme' showed the lowest susceptibility to tephritids in terms of puparia per fruit. The variability of infestation among the guava genotypes and the reasons for increasing fruit fly infestations along the time were discussed.     

Susceptibility of six Arabic coffee cultivars to fruit flies (Diptera: Tephritoidea) under shaded and unshaded organic management in Valença, State of Rio de Janeiro, Brazil

The infestation indices by fruit flies were determined for six cultivars of Coffea arabica L. in shaded and unshaded systems under organic management. The experiment was set in a completely randomized design with a split-split-plot arrangement and four replicates. A 250g-sample of maturing fruits per plot was harvested in May 2005. The cultivars Icatu Amarelo and Catucaí Amarelo were the least susceptible to attack by tephritids in both systems. As for lonchaeids, Oeiras, Catucaí Amarelo and Catuaí Vermelho were the least susceptible cultivars in the shaded system, and there was no difference among the cultivars in the unshaded system. The following tephritid species were obtained: Ceratitis capitata (Wiedemann), Anastrepha fraterculus (Wiedemann) and A. sororcula Zucchi (Tephritidae). Lonchaeids were represented by Neosilba bifida Strikis & Prado, N. certa (Walker), N. glaberrima (Wiedemann), N. pendula (Bezzi), N. pseudopendula (Korytkowski and Ojeda), Dasiops rugifrons Hennig, Neosilba n.sp.10 and Neosilba n.sp.14.     

Morphometry and distribution of sensilla on the antennae of Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae)

Antennal sensilla of Anastrepha fraterculus (Wied.) were examined using scanning electron microscopy. In the flagellum, there are trichoid, basiconic, clavate type I and II, and styloconic sensilla and microtrichia. Only microtrichiae and chaetica sensilla were observed in the scape and pedicel. The number of sensilla in the flagellum was similar between sexes. At the apex there was a higher density of trichoid and an absence of clavate sensilla, while basiconic sensilla were more abundant in the proximal region.     

Digestion of bacteria and the role of midgut lysozyme in some insect larvae.

1. Lysozyme is absent from tissues other than the midgut in the drug-feeding larvae of Musca domestica (Diptera, Cyclorrhapha, Muscidae) and in the fruit-feeding larvae of Anastrepha fraterculus (Diptera, Cyclorrhapha, Tephritidae), whereas in the detritus-feeding larvae of Trichosia pubescens (Diptera, Nematocera, Sciaridae) lysozyme is only found in the hemolymph and in the fat body. 2. A. fraterculus larvae have a midgut region with a luminal pH of 3.4, and display a pepstatin-inhibited acid proteolytic activity which has a spec. act. (7.2 U/mg protein) similar to that of M. domestica. 3. The midgut lysozyme from M. domestica and A. fraterculus is more active (high ionic strength) at pH 3.5 than at pH 6.0, the contrary being true for a midgut chitinase. 4. The results suggest that the adaptations to digest bacteria in insects are similar to those in vertebrate foregut fermenters, and that these characteristics were probably present in the Cyclorrhapha ancestor, but not in the Diptera ancestor.     

Anastrepha ludens and Anastrepha serpentina (Diptera: Tephritidae) do not infest Psidium guajava (Myrtaceae), but Anastrepha obliqua occasionally shares this resource with Anastrepha striata in nature

This study examined whether economically important fruit fly species Anastrepha ludens (Loew), Anastrepha serpentina (Wiedemann), and Anastrepha obliqua (Macquart) (Diptera: Tephritidae) may opportunistically exploit guavas, Psidium guajava L. (Myrtaceae), growing near preferred natural hosts. We collected 3,459 kg of guavas and 895 kg of other known host species [sour orange, Citrus aurantium L.; grapefruit, Citrus paradisi Macfadyen; mango, Mangifera indica L.; white sapote, Casimiroa edulis La Llave and Lex.; sapote, Pouteria sapota (Jacq.); sapodilla, Manilkara zapota L.; and wild plum, Spondias purpurea L. and Spondias mombin L.] along an altitudinal gradient over a 4-yr period (2006-2009). Plants were growing in sympatry in 23 localities where the guavas are usually infested in the state of Veracruz, M6xico. The guava samples yielded 20,341 Anastrepha spp. pupae in total (overall mean, 5.88 pupae per kg of fruit). Confirming previous reports, Anastrepha fraterculus (Wiedemann) and Anastrepha striata (Schiner) were found heavily infesting guavas in Veracruz. Importantly, although we did not find evidence that A. ludens and A. serpentina are able to attack this valuable commodity, we document for the first time in the agriculturally important state of Veracruz that P. guajava is an alternative natural host plant of A. obliqua. We recovered two fruit in the mango-growing locality of la Vibora, Tlalixcoyan, that harbored larvae of A. striata and A. obliqua. This finding has important practical implications for management of A. obliqua. Over the entire altitudinal gradient, when individual fruit infestation was examined, a dynamic pattern of species dominance was unveiled with guavas growing below 800 m above sea level mainly attacked by A. striata and a progressive replacement with increasing altitude by A. fraterculus. Interestingly, most individual fruit examined (97%) harbored a single species of fruit fly, a finding that may be taken as evidence of competitive displacement among sympatric species of fruit flies. Based on this study and previously published work by us on this topic, we conclude that literature reports indicating that A. ludens and A. serpentina infest guavas under field conditions should be questioned.     

A morphometric and molecular study of Anastrepha pickeli Lima (Diptera: Tephritidae)

This study investigated the level of morphometric and genetic variability among populations of Anastrepha pickeli Lima from several localities in Brazil, one locality in Bolivia and one in Paraguay. Traditional and geometric morphometric analyses were used, as well as sequencing of a fragment of the cytochrome oxidase gene (COI). Six variables were measured from the aculeus for traditional morphometric analysis and 14 landmarks from the right wing were used for geometric analysis, using 10 specimes/population. The aculeus tip length, aculeus width at the end of the cloaca opening, and the serrate part length contributed with 62.7% for grouping. According to the results from traditional morphometry, there was no significant difference, but the multivariate tests showed that the canonical variables were statistically significant, indicating a difference in the wing conformation among populations. Molecular phylogenetic analysis indicated that the populations clustered into three clades and revealed a high level of genetic variation within A. pickeli populations from various geographic regions. Anastrepha pickeli populations differed among them according to the methods used in this study, showing incongruence among the methods used.     

Karyotypic and molecular identification of laboratory stocks of the South American fruit fly Anastrepha fraterculus (Wied) (Diptera: Tephritidae)

The taxonomic status of the tephritid pest Anastrepha fraterculus (Wied.) is a controversial subject, mainly because of misinterpretation of the observed genetic variation. In this work, the different karyotypes and DNA polymorphism of a geographically defined population from Northeastern Argentina were studied, using derived stocks maintained in the laboratory during 25 generations. The karyotypes were analyzed using C-banding and N-banding techniques, while DNA analysis was performed through the DNA polymerase chain reaction (PCR). The variants isolated from both the wild Montecarlo population and the derived laboratory stocks were fully compatible and are present in other wild populations from South Brazil (lat 31 degrees 30' S) to Mid-Argentina (lat 34 degrees 30' S). Single-pair crosses among stocks carrying different chromosomal variants demonstrated the absence of isolation barriers. The polymorphic fragments isolated by RAPDs/PCR showed polymorphisms among stocks whereas the analysis of rDNA ITS1 exhibit a unique ITS1 length. Our results seem to indicate that all the examined variants belong to a single species with extended polymorphism and therefore do not support the hypothesis that the extended chromosomal polymorphism in A. fraterculus implies the existence of a complex of cryptic species.     

Factors affecting Anastrepha fraterculus female receptivity modulation by accessory gland products

In the context of the sterile insect technique (SIT), mass-rearing and male irradiation are imperative. Post-teneral treatments such as the addition of protein in adult's male diet and male hormonal treatment are used to improve sexual performance and to accelerate sexual maturation. In this work we investigated the effect of male accessory glands products (AGPs) on female receptivity of the South American fruit fly Anastrepha fraterculus (Wiedemann), and the effect of strain rearing history, male irradiation, male diet and hormonal treatment on AGPs. Injections of aqueous extracts of male accessory glands into the abdomen of females reduced their receptivity. The AGPs from laboratory males were more effective in inhibiting female receptivity, compared to AGPs from wild males, irrespective of females' origin. The AGPs from fertile males were more effective than AGPs from sterile males. The AGPs from protein-fed males were more effective than AGPs from sugar-fed males. Finally, the AGPs of males treated with juvenile hormone were less effective in inhibiting female receptivity than AGPs of untreated males. We conclude that inhibition of sexual receptivity of A. fraterculus mated females is mediated by products in male accessory gland's and the way that these products act vary widely according to the effect of extrinsic factors. We discuss the results in the perspective of the SIT application for A. fraterculus.     

Positive and purifying selection influence the evolution of doublesex in the Anastrepha fraterculus species group

The gene doublesex (dsx) is considered to be under strong selective constraint along its evolutionary history because of its central role in somatic sex differentiation in insects. However, previous studies of dsx used global estimates of evolutionary rates to investigate its molecular evolution, which potentially miss signals of adaptive changes in generally conserved genes. In this work, we investigated the molecular evolution of dsx in the Anastrepha fraterculus species group (Diptera, Tephritidae), and test the hypothesis that this gene evolved solely by purifying selection using divergence-based and population-based methods. In the first approach, we compared sequences from Anastrepha and other Tephritidae with other Muscomorpha species, analyzed variation in nonsynonymous to synonymous rate ratios (dN/dS) in the Tephritidae, and investigated radical and conservative changes in amino acid physicochemical properties. We show a general selective constraint on dsx, but with signs of positive selection mainly in the common region. Such changes were localized in alpha-helices previously reported to be involved in dimer formation in the OD2 domain and near the C-terminal of the OD1 domain. In the population-based approach, we amplified a region of 540 bp that spanned almost all of the region common to both sexes from 32 different sites in Brazil. We investigated patterns of selection using neutrality tests based on the frequency spectrum and locations of synonymous and nonsynonymous mutations in a haplotype network. As in the divergence-based approach, these analyses showed that dsx has evolved under an overall selective constraint, but with some events of positive selection. In contrast to previous studies, our analyses indicate that even though dsx has indeed evolved as a conserved gene, the common region of dsx has also experienced bouts of positive selection, perhaps driven by sexual selection, during its evolution.     

Incipient speciation revealed in Anastrepha fraterculus (Diptera; Tephritidae) by studies on mating compatibility, sex pheromones, hybridization, and cytology

It has long been proposed that the nominal species Anastrepha fraterculus is a species complex and earlier studies showed high levels of pre-zygotic isolation between two laboratory strains from Argentina and Peru. Further experiments were carried out on the same populations and on their reciprocal hybrids, including pre- and post-zygotic isolation studies, pheromone analysis, and mitotic and polytene chromosome analysis. A high level of pre-zygotic isolation had been maintained between the parental strains despite 3 years of laboratory rearing under identical conditions. The level of pre-zygotic isolation was reduced in matings with hybrids. There were also differences in other components of mating behaviour. There were quantitative and qualitative differences in the sex pheromone of the two strains with the hybrids producing a mixture. The pre-zygotic isolation barriers were complemented by high levels of post-zygotic inviability and sex ratio distortion, most likely not due to Wolbachia , although there was evidence of some cytoplasmic factor involved in sex ratio distortion. Analysis of polytene chromosomes revealed a high level of asynapsis in the hybrids, together with karyotypic differences between the parental strains. The combined results of the present study indicate that these two strains belong to different biological entities within the proposed A. fraterculus complex.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 97 , 152–165.