Lucie Vaníčková

Pheromone Analyses of the Anastrepha fraterculus (Diptera: Tephritidae) Cryptic Species Complex

ABSTRACT The South American fruit fly Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) cryptic species complex is presently one of the most studied pest models in terms of speciation and population mating compatibility. The improvement of pest-control techniques has strongly relied on successful implementation of laboratory strains into wild populations. Pheromone communication plays an important role in the mating process in the South American fruit fly. Therefore, the main goal of the present study was to investigate the pheromone composition of 7 different populations, originating from geographically distant locations in Brazil and Argentina. Fourteen volatile compounds were identified in calling male emanations by GC×GC/TOF-MS and the data obtained were subsequently analyzed by multivariate statistics. The pheromone composition varied both quantitatively and qualitatively among the studied populations. These results will serve as the basis for further electrophysiological analyses.

Resolution of three cryptic agricultural pests (Ceratitis fasciventris, C. anonae, C. rosa, Diptera: Tephritidae) using cuticular hydrocarbon profiling.

Discrimination of particular species within the species complexes of tephritid fruit flies is a very challenging task. In this fruit-fly family, several complexes of cryptic species have been reported, including the African cryptic species complex (FAR complex). Cuticular hydrocarbons (CHCs) appear to be an excellent tool for chemotaxonomical discrimination of these cryptic species. In the present study, CHC profiles have been used to discriminate among three important agricultural pests from the FAR complex, Ceratitis fasciventris, Ceratitis anonae and Ceratitis rosa. Hexane body surface extracts of mature males and females were analyzed by two-dimensional gas chromatography with mass spectrometric detection and differences in CHC profiles between species and sexes tested through multivariate statistics and compared with species identification by means of microsatellite markers. Quantitative as well as qualitative CHC profile differences between sexes and species are reported. The CHC profiles consisted of a mixture of linear, internally methyl-branched and mono-, di- and tri-unsaturated alkanes. Twelve compounds were pinpointed as potential chemotaxonomical markers. The present study shows that presence or absence of particular CHCs might be used in the chemical diagnosis of the FAR complex. Moreover, our results represent an important first step in the development of a useful chemotaxonomic tool for cryptic species identification of these important agricultural pests.

Are the wild and laboratory insect populations different in semiochemical emission? The case of the medfly sex pheromone.

The medfly (Ceratitis capitata) is one of the major agricultural pests controlled through sterile insect technique (SIT) programs. We studied the chemical composition of the volatiles released by calling males from one laboratory and two wild C. capitata populations using two-dimensional gas chromatography with time-of-flight mass spectrometric detection (GC × GC/TOFMS) and gas chromatography with electroantennographic detection (GC-EAD). Multivariate data analyses revealed significant differences in the quantitative and qualitative composition of male chemical emanations between the three populations. The GC-EAD analyses of the male emanation of three C. capitata populations revealed 14 antenally active compounds. The volatiles isomenthone, β-pinene, ethyl octanoate, indole, geraniol, bornyl acetate, geranyl acetone, and (E)-caryophyllene are newly reported EAD active constituents of the male pheromone. GC-EAD analyses of the laboratory population indicated that the males and females of C. capitata possess comparable sensitivity to male-produced volatiles. Our results are relevant to the development of a pheromone-based monitoring system and also to the SIT control program.

Intraspecific variation of cuticular hydrocarbon profiles in the Anastrepha fraterculus (Diptera: Tephritidae) species complex

The goal of this study was to define whether cuticular hydrocarbons (CHs) could be used for taxonomic determination of putative species hidden in the Anastrepha fraterculus cryptic species complex, widespread from Argentina to Mexico. Recently, increasing evidence of phenotypic and genetic variability has resulted in the characterization of eight morphotypes within this complex. The CH profiles of six A. fraterculus populations from Argentina, Brazil, Peru, Colombia and Mexico were analysed in this study by two‐dimensional gas chromatography coupled with mass spectrometry. In parallel, multiple factorial analyses were used to elucidate population structures. Vector populations segregated into four distinct groups. The analysis demonstrated that the studied populations from Peru, Argentina and southern Brazil (Vacaria) might be classified in accordance with the earlier division of the A. fraterculus complex into Peruvian, and Brazilian‐1 cryptic species, using the specific CH profiles. Population from south‐eastern Brazil (Piracicaba) formed separated group. Mexican and Andean (Colombian) putative species had similar CH signatures, when compared to each other.

Identification of male-borne attractants in Anastrepha fraterculus (Diptera: Tephritidae)

The South American fruit fly is one of the most destructive polyphagous pests in South America. In this species, males gathered in aggregations emit volatiles that attract females; however, the compounds involved in this task remain unknown. In this study, we investigated the chemical composition of the volatile blend emitted by males aiming to identify the specific compounds within this blend that elicit behavioral responses in conspecific females. For this purpose, we performed chemical and electrophysiological analyses and bioassays. The chemical analyses revealed the presence of 29 compounds in headspace samples of A. fraterculus males, of which six compounds, i.e. α-pinene, limonene, (Z)-3-nonen-1-ol, (E,Z)-3,6-nonadien-1-ol, α-farnesene and (S,S)-(−)-epianastrephin, triggered antennal depolarization in conspecific females. In laboratory bioassays, five out of eight synthetic compounds tested individually elicited more behavioral responses than a hexane control, but only the synthetic mixture composed of all EAD-active compounds triggered behavioral responses in females similar to the responses to the headspace samples of conspecific males. In an experiment under semi-natural conditions, the synthetic mixture was more attractive to females than a hexane control and equally attractive to headspace extracts of males. This study reports the identification of male volatile compounds that act as attractant for A. fraterculus females, which may be useful for the control of this pest in infested orchards.

Current knowledge of the species complex Anastrepha fraterculus (Diptera, Tephritidae) in Brazil

Abstract The study of the species complex Anastrepha fraterculus (Af complex) in Brazil is especially important in a taxonomical, evolutionary and pest management context, because there are evidences that some of them may occur in sympatry. In this review, we analyzed the main results supporting evidences that three cryptic species occur in Brazil. The taxonomical and phylogenetic relationships based on eggshell morphology, adult morphometrics, as well as cytotaxonomy and genetic differentiations are discussed. We also review available information on sexual behavior including acoustic communication of males during courtship and sexual incompatibility; and chemical signals involved in the communication between sexes, with a special focus on sex pheromones. We examined the role of long- and short-range pheromones (male-produced volatiles and cuticular hydrocarbons, respectively), their implications in sexual isolation, and their possible use for chemotaxonomic differentiation of the putative species of the Af complex.

Cuticular hydrocarbons corroborate the distinction between lowland and highland Natal fruit fly (Tephritidae, Ceratitis rosa) populations.

Abstract The cuticular hydrocarbons (CHs) and morphology of two Ceratitis rosa Karsch (Diptera: Tephritidae) populations, putatively belonging to two cryptic taxa, were analysed. The chemical profiles were characterised by two-dimensional gas chromatography with mass spectrometric detection. CHs of Ceratitis rosa that originated from the lowlands and highlands of Kenya comprised of n-alkanes, monomethylalkanes, dimethylalkanes and unsaturated hydrocarbons in the range of the carbon backbone from C14 to C37. Hydrocarbons containing C29, C31, C33 and C35 carbon atoms predominated in these two populations. 2-Methyltriacontane was the predominant compound in both populations. Quantitative differences in the distribution of hydrocarbons of different chain lengths, mainly the C22, C32, C33 and C34 compounds of these two populations, were observed despite indistinct qualitative differences in these hydrocarbons. Morphological analyses of male legs confirmed that the flies belong to different morphotypes of Ceratitis rosa previously labelled as R1 and R2 for lowland and highland populations, respectively. A statistical analysis of the CH compositions of the putative R1 and R2 species showed distinct interspecific identities, with several CHs specific for each of the lowland and highland populations. This study supports a hypothesis that the taxon Ceratitis rosa consists of at least two biological species.

Pheromone Communication in Anastrepha obliqua (Diptera: Tephritidae): A Comparison of the Volatiles and Salivary Gland Extracts of Two Wild Populations

Abstract The West Indian fruit fly Anastrepha obliqua Macquart (Diptera: Tephritidae) is one of the major pests on mango (Mangifera indica L.; Sapindales: Anacardiaceae) and starfruit (Averrhoa carambola L.; Oxalidales: Oxalidaceae) crop plantations in Brazil. Pheromone communication inter alia plays an important role in fruit-fly courtship behavior. In order to highlight the site of pheromone synthesis, we identified and compared the volatiles from the aeration extracts of calling males with the volatiles produced by their salivary glands in 2 wild populations of A. obliqua collected from mangoes and starfruits. In addition, we performed a series of bioassays to compare the biological significance of both extracts. In total, 36 volatile compounds were identified, with 8 of them being shared by the 2 populations and the 2 extract types. Linalool and &agr;-copaene were exclusively found in the aeration extract while ethyl heptanoate, methyl octanoate, and 1-nonanol were detected only in the salivary-gland extracts. The chemical profiles of the volatiles from the aeration extracts and from the salivary-gland extracts differed significantly between the 2 populations as well as the chemical profiles of both extracts within each population. The quantities of the 8 shared compounds generated a variability of more than 60% in the mango population and 80% in the starfruit population. The similarities observed between the chemical profiles of the aeration extracts and the salivary-gland extracts suggest that the latter could be the storage site and probably also the production site of some pheromone components in this fruit-fly species. This hypothesis is supported by the comparable biological activities of both extracts in terms of their attractiveness for conspecific females.

Characterisation of the chemical profiles of Brazilian and Andean morphotypes belonging to the Anastrepha fraterculus complex (Diptera, Tephritidae).

Abstract Fruit fly sexual behaviour is directly influenced by chemical and non-chemical cues that play important roles in reproductive isolation. The chemical profiles of pheromones and cuticular hydrocarbons (CHs) of eight fruit fly populations of the Andean, Brazilian-1 and Brazilian-3 morphotypes of the Anastrepha fraterculus cryptic species complex originating from Colombia (four populations) and Brazil (four populations) were analysed using two-dimensional gas chromatography with mass spectrometric detection. The resulting chemical diversity data were studied using principal component analyses. Andean morphotypes could be discriminated from the Brazilian-1 and Brazilian-3 morphotypes by means of male-borne pheromones and/or male and female CH profiles. The Brazilian-1 and Brazilian-3 morphotypes were found to be monophyletic. The use of chemical profiles as species- and sex-specific signatures for cryptic species separations is discussed.

Sexual chemoecology of mosquitoes (Diptera, Culicidae): Current knowledge and implications for vector control programs

Mosquitoes (Diptera: Culicidae) act as vectors of medical and veterinary importance, due to their ability to transmit many pathogens and parasites. Renewed interest has been recently devoted to the potential of sterile insect technique (SIT) for mosquito suppression. However, the success of the SIT is mostly dependent on the ability of sterile males to compete for mates with the wild ones in the field. Nevertheless, little is known on the sexual chemical ecology of mosquitoes, with special reference to the role of chemical signals in males. We reviewed the current knowledge on mosquito sexual chemical ecology and other key cues affecting courtship and mating behavior. The information available on the aggregation and sex pheromones in mosquito males is rather limited. To the best of our knowledge, the components of the aggregation pheromone stimulating swarming mechanisms have been fully characterized only for Aedes aegypti, while evidence for aggregation pheromones in other mosquito species remains elusive. Further research on this issue is needed, as well as to dissect the relative importance of visual (with special reference to swarming landmarks), vibrational, olfactory and tactile cues perceived during swarming and mate. On the other hand, more knowledge is available for cuticular hydrocarbons, which modulate mating behavior in several species of economic importance. These compounds, coupled with volatile aggregation components, have potential interest for the development of monitoring and trapping systems. In addition, the analyses of cuticular hydrocarbons are essential for discrimination between closely related mosquito species and/or populations.