Evaluation of trap types and food attractants for Rhagoletis cerasi (Diptera: Tephritidae)

Trapping experiments were conducted during the period of flight activity of the cherry fruit fly Rhagoletis cerasi L. in the area of Thessaloniki, northern Greece, during the years 1993-1997 to test several traps alone and in combination with different food attractants. Yellow sticky-coated visual traps were more effective than McPhail-type traps baited with different food attractants. Of the visual traps, the most effective was the yellow Rebell trap. The Rebell trap, provided with a dispenser containing a slow release formulation of ammonium acetate attached to the lower part of the trap, was found to be the most effective of all treatments tested, capturing approximately 50% more R. cerasi flies than the Rebell trap without any attractant. Ammonium carbonate dispensers did not increase the performance of Rebell traps. More mature females were captured in Rebell traps baited with or without ammonium acetate than in McPhail-type traps baited with ammonium acetate. This study demonstrates that Rebell traps baited with an ammonium acetate dispenser can provide a more effective tool for monitoring and mass trapping of R. cerasi than the currently used unbaited Rebell traps.     

Genetic and cytogenetic analysis of the fruit fly Rhagoletis cerasi (Diptera: Tephritidae).

The European cherry fruit fly, Rhagoletis cerasi, is a major agricultural pest for which biological, genetic, and cytogenetic information is limited. We report here a cytogenetic analysis of 4 natural Greek populations of R. cerasi, all of them infected with the endosymbiotic bacterium Wolbachia pipientis. The mitotic karyotype and detailed photographic maps of the salivary gland polytene chromosomes of this pest species are presented here. The mitotic metaphase complement consists of 6 pairs of chromosomes, including one pair of heteromorphic sex chromosomes, with the male being the heterogametic sex. The analysis of the salivary gland polytene complement has shown a total of 5 long chromosomes (10 polytene arms) that correspond to the 5 autosomes of the mitotic nuclei and a heterochromatic mass corresponding to the sex chromosomes. The most prominent landmarks of each polytene chromosome, the "weak points", and the unusual asynapsis of homologous pairs of polytene chromosomes at certain regions of the polytene elements are also presented and discussed.     

Prolonged pupal dormancy is associated with significant fitness cost for adults of Rhagoletis cerasi (Diptera: Tephritidae)

In temperate areas, dormancy (diapause and/or quiescence) enables herbivorous insect species to persist and thrive by synchronizing growth and reproduction with the seasonal phenology of their host plants. Within-population variability in dormancy increases survival chances under unpredictable environmental changes. However, prolonged dormancy may be costly, incurring trade-offs in important adult fitness traits such as life span and reproduction. We used the European cherry fruit fly, Rhagoletis cerasi, a stenophagous, univoltine species that overwinters in the pupal stage for usually one or more years to test the hypotheses that prolonged dormancy of pupae has trade-offs with body size, survival and reproduction of the resulting adults. We used two geographically isolated populations of R. cerasi to compare the demographic traits of adults obtained from pupae subjected to one or two cycles of warm-cold periods (annual and prolonged dormancy respectively). Regardless of population, adults from pupae that experienced prolonged dormancy were larger than counterparts emerging within 1year. Prolonged dormancy did not affect adult longevity but both lifetime fecundity and oviposition were significantly decreased. Extension of the life cycle of some individuals in R. cerasi populations in association with prolonged dormancy is likely a bet-hedging strategy.     

Visual responses of apple maggot flies, Rhagoletis pomonella (Diptera: Tephritidae): Orchard studies

Apple maggot flies, Rhagoletis pomonella, were found to be highly responsive to visual stimuli, with maximum responses dependent upon color, shape, and size characteristics of the stimulus. Each sex had a significant preference for yellow 30×40 cm rectangles over green, orange, red, blue, violet, black, white and clear rectangles of the same size. On the other hand, each sex had a significant preference for red, blue, violet, dark organge, and black spheres 7.5 cm in diameter over green, light orange, yellow, white, and clear spheres of the same size. Both sexes had a significant preference for 7.5 cm spheres over equivalent-sized cubes, cylinders and rectangles. As the diameter of a sphere was increased from 7.5 to 45 cm, there was an orderly and significant decrease in the attractiveness of those that were darkcolored (red) but a progressive and significant increase in the attractiveness of those that were yellow. The flies did not congregate on trees with apples or respond strongly to 7.5 cm, darkcolored spheres until they were sexually mature (at least in the case of females). Relative to other colors, females were often significantly more attracted than males to yellow. The attractiveness of an olfactory stimulus eliciting feeding-type reactions was enhanced to a substantial degree when employed in conjunction with a 30×40 cm yellow rectangle, but to only a very slight degree when in conjunction with a 7.5 cm red sphere.Evidence presented suggests the following explanation for these findings. The flies are attracted to large surfaces of yellow because they react to yellow, on the basis of true color discrimination, as if it were foliage on which to find food. On the other hand, they are attracted to small, dark-colored spheres because they react to such spheres as they react to apples, which are the oviposition site and which also appear to serve as a rendezvous site for the sexes in mating activity. Once the flies have arrived on apple trees, they detect apples solely through vision. Red, blue, violet, dark orange, and black small spheres are preferred over small spheres of other colors on the basis that they stand out in strongest contrast against the background and not on the basis of true color discrimination. Large, dark-colored spheres are unattractive because the range of sphere sizes eliciting positive responses is near the size of an apple.These findings have proven useful in designing effective devices for sampling orchard populations of the flies and may also prove useful in fly control.

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Zusammenfassung Imagines von Rhagoletis pomonella reagieren positiv auf visuelle Reize, wobei Farbe, Form und Grösse des Stimulus von ausschlaggebender Bedeutung sind. Beide Geschlechter zeigten in den Versuchen eine signifikante Präferenz für gelbe Rechtecke (30×40 cm), die sie häufiger anflogen als grüne, orange, rote, blaue, violette, schwarze, weisse und farblose Flächen gleicher Grösse. Ebenso zeigte es sich, dass Kugeln von 7.5 cm Durchmesser von den Fliegen eindeutig Kuben, Zylindern und Rechtecken gleicher Grössenordnung für den anflug bevorzugt wurden. Mit zunehmendem Durchmesser der Kugeln von 7.5 cm bis 45 cm konnte eine abnehmende Anziehung von dunkel gefärbten Kugeln, jedoch eine zunehmende Anziehung von gelben Kugeln auf die Fliegen beobachtet werden. Vor der Geschlechtsreife reagierten die Weibchen nicht in ausgeprägtem Maße auf die 7.5 cm dunkel gefärbte grossen Kugeln, sondern wurden mehr als die Männchen von Gelb angezogen. Die Wirkung eines Nahrungsköders wurde in Kombination mit einer gelben Fläche von 30×40 cm signifikant erhöht, während die Kombination des Köders mit einer roten Kugel von 7.5 cm Durchmesser die Anziehung von Weibchen nur unwesentlich erhöhte.Die Resultate lassen sich damit erklären, dass die Fliegen durch gelbe Flächen angezogen werden, weil sie dank Diskriminierung der Farbe auf Gelb reagieren als wäre es Blattwerk, auf dem sich die Nahrungsquellen befinden. Auf der anderen Seite werden sie von dunklen Kugeln in den Dimensionen eines Apfels angezogen, da im Freiland Äpfel Orte für die Eiablage und offenbar Treffpunkte für die Geschlechter darstellen. Sobald die Fliegen den Apfelbaum beflogen haben, orientieren sie sich allein mit dem Gesichtssinn gegen die Früchte. Kleinere rote, blaue, dunkel orange, violette und schwarze Kugeln werden anders gefärbten vorgezogen, weil sie den grössten Kontrast gegenüber dem Hintergrund bilden und nicht auf Grund echter Farbdiskriminierung. Grosse dunkle Kugeln verlieren ihre Wirkung auf die Fliegen, weil nur Kugeln in der Grössenordnung eines Apfels positive Reaktionen auslösen.Diese Befunde haben sich als nützlich erwiesen in der Ausarbeitung von Sammelmethoden für Freilandpopulationen und dürften auch in der Bekämpfung der Apfelfliegen Anwendung finden.

     

Habitat-specific flight period in the cherry fruit fly Rhagoletis cingulata (Loew) (Diptera: Tephritidae)

Flight periods of the cherry fruit fly, Rhagoletis cingulata (Loew), were compared in the major sweet and tart cherry-growing regions of Michigan, among neglected orchards, managed orchards, and natural areas containing the ancestral host, black cherry. Traps were deployed from early June to late September 2005 and 2006. Captures indicated that cherry fruit fly has an early flight (June-July) in neglected orchards, a mid-season flight peaking immediately after harvest (June-August) in managed orchards, and an extended flight covering most of the season (June-September) in natural areas. We found that the period of fruit infestation mirrored the flight period in neglected and managed orchards. In natural areas, we found infestation late in the season only. The relative emergence periods for adults reared from pupae collected from the three habitats and maintained under the same conditions coincided with adult flight periods for each habitat. We also studied factors related to fruit availability that may have a role in shaping the flight periods. Fruit abundance decreased rapidly early in the season in neglected orchards, whereas in managed orchards, fruit left after harvest remained on the trees until late August. Measurements of fruit size and skin firmness revealed that fly activity in neglected and managed orchards began immediately after fruit increased in size and skin firmness decreased, whereas in natural areas, the flight began before fruit matured. In managed orchards, fruit harvest and insecticide sprays likely maintain the late flight period of resident fly populations by preventing the use of fruit earlier in the season. However, a significant proportion of these resident flies may still emerge before harvest and increase the risk of costly fruit infestation.     

Evidence for a recent horizontal transmission and spatial spread of Wolbachia from endemic Rhagoletis cerasi (Diptera: Tephritidae) to invasive Rhagoletis cingulata in Europe

The widespread occurrence of Wolbachia in arthropods and nematodes suggests that this intracellular, maternally inherited endosymbiont has the ability to cross species boundaries. However, direct evidence for such a horizontal transmission of Wolbachia in nature is scarce. Here, we compare the well‐characterized Wolbachia infection of the European cherry fruit fly, Rhagoletis cerasi, with that of the North American eastern cherry fruit fly, Rhagoletis cingulata, recently introduced to Europe. Molecular genetic analysis of Wolbachia based on multilocus sequence typing and the Wolbachia surface protein wsp showed that all R. cingulata individuals are infected with wCin2 identical to wCer2 in R. cerasi. In contrast, wCin1, a strain identical to wCer1 in R. cerasi, was present in several European populations of R. cingulata, but not in any individual from the United States. Surveys of R. cingulata from Germany and Hungary indicated that in some populations, the frequency of wCin1 increased significantly in just a few years with at least two independent horizontal transmission events. This is corroborated by the analysis of the mitochondrial cytochrome oxidase II gene that showed association of wCin1 with two distinct haplotypes in Germany, one of which is also infected with wCin1 in Hungary. In summary, our study provides strong evidence for a very recent inter‐specific Wolbachia transmission with a subsequent spatial spread in field populations.     

Hidden Wolbachia diversity in field populations of the European cherry fruit fly, Rhagoletis cerasi (Diptera, Tephritidae)

The European cherry fruit fly Rhagoletis cerasi has been a field model for cytoplasmic incompatibility since the mid 1970s. Two Wolbachia strains were detected in this tephritid species and w Cer2 was described as the CI inducing agent dividing European populations into two unidirectional incompatible groups, i.e. southern females produce viable offspring with northern males, whereas the reciprocal cross results in incompatibility. We detected three new Wolbachia strains by sequencing a multitude of plasmids derived from Wolbachia surface protein gene ( wsp ) polymerase chain reaction (PCR) products. Strain-specific primers were developed allowing individual diagnosis without need for cloning. Hybridization of specific PCR products with a wsp oligonucleotide enhanced the detection limit significantly and revealed the presence of low-titre infections in some strains, in different ontogenetic stages and in adults of different age. We then performed a survey of strain prevalence and infection frequency in eight European regions. w Cer1 was fixed in all populations, whereas w Cer2 was detected only in the South. w Cer3 frequency was the lowest without a clear distribution pattern. The abundance of w Cer4 was homogenous across Europe. Like w Cer2, w Cer5 showed significant differences in spatial distribution. Our new findings of previously undetected and recombinant Wolbachia strains in R. cerasi reveal a major caveat to the research community not to overlook hidden Wolbachia diversity in field populations. Low-titres and geographical variability in Wolbachia diversity are expected to influence the outcome of Wolbachia population dynamics and Wolbachia- based insect population control and may create invasion barriers for expanding and artificially introduced Wolbachia strains.     

Wolbachia infections and superinfections in cytoplasmically incompatible populations of the European cherry fruit fly Rhagoletis cerasi (Diptera,Tephritidae)

Wolbachia is an obligately intracellular, maternally inherited bacterium which has been detected in many arthropods. Wolbachia infections disperse in host populations by mechanisms such as cytoplasmic incompatibility (CI). CI leads to embryonic mortality which occurs when infected males mate with uninfected females or females with a different Wolbachia strain. Populations of the European cherry fruit fly Rhagoletis cerasi (Diptera, Tephritidae) were found to be infected by two different Wolbachia strains, wCer1 and wCer2. Superinfections with both strains occurred throughout southern and central Europe and infections with wCer1 were found in northern, western and eastern Europe. Strong unidirectional CI between European populations of R. cerasi were first reported in the 1970s. From the conformity in the recent geographical distribution of the Wolbachia infections and the CI expression patterns found 25 years ago it was deduced that wCer2 potentially causes CI in R. cerasi. The comparison of the geographical distributions indicated that wCer1 + 2 must have spread into wCer1-infected populations in some areas. In other regions, a spread of wCer1 + 2 was probably prevented by dispersal barriers. There, a sharp transition from infected to superinfected populations suggested regional isolation between wCer1 and wCer1 + 2-infected populations.     

Potential geographical distribution of Rhagoletis pomonella (Diptera: Tephritidae) in China

Abstract Apple maggot fly, Rhagoletis pomonella (Walsh) is a major pest causing considerable economic losses of fruits in North America. During the development of international trade, apple maggot fly has become a threat to Chinese agriculture. In this study, CLIMEX and ArcGIS were used to predict the potential geographical distribution of apple maggot fly in China. The parameters used in CLIMEX for apple maggot fly were derived from ecological data and the present geographical distribution of apple maggot fly in North America. Then the potential distribution map in China was presented based on the adjusted values of these parameters. The results showed that apple maggot fly has a wide potential distribution area in China; 47.5% of 748 meteorological stations presented high or medium suitability of pest establishment. These high suitable stations are mainly located in northeast, southwest and northwest of China, such as Liaoning, Shandong, Gansu and Shaanxi Provinces. These areas are also the central regions of apple, pear and peach production in China. Two hundred and twenty‐five stations (30.1%) in western and southern China, such as Tibet, Qinghai, Guangdong, Guangxi, Hainan and Taiwan, were unsuitable for establishment of apple maggot fly populations. In order to prevent the introduction of apple maggot fly in China, the present plant quarantine measures should be enhanced, especially in the areas with high suitability for the presence of apple maggot fly.     

Sex-specific glands in tephritid fruit flies of the genera Anastrepha,Ceratitis, Dacus and Rhagoletis (Diptera : Tephritidae)

The objectives of this work were to systematically examine males and females of tephritid fruit flies from the genera Anastrepha, Ceratitis, Dacus and Rhagoletis for the presence of possible sex pheromone glands, and to determine whether particular glands may be characteristic of certain genera. Flies from the 4 genera were examined from paraffin embedded histological sections. Pleural epidermal glands and sexually dimorphic, enlarged salivary glands were found in males, but not in females, of 7 species of Anastrepha and in 2 species of Ceratitis. Females of these species do not develop the pleural epidermal glands at all, and although they have salivary glands, the glands do not have the large glandular mass of tissue that is found in males. The pleural glands were similar in all the males, and consisted of a single layer of enlarged, columnar epidermal cells in the pleural region of abdominal segments 3, 4 and 5. The salivary glands of males were divided into 4 types based upon structure.Pleural glands and enlarged salivary glands were searched for, but were not found, in 2 species of Rhagoletis and in 3 species of Dacus. Anal glands, opening externally on the last segment, were confirmed in C. capitata, and were found in C. rosa. Anal glands were not found in any of the other species. A diverticulum from the rectum, the rectal pouch or gland, was confirmed in D. dorsalis, D. cucurbitae and D. tryoni as already published, but this gland was not found in species of other genera. Growth of the pleural glands and salivary glands in males of A. ludens, A. fraterculus and C. capitata was described as a function of age of flies.     

Obligate annual and successive facultative diapause establish a bet‐hedging strategy of Rhagoletis cerasi (Diptera: Tephritidae) in seasonally unpredictable environments

To cope with temporal and spatial heterogeneity of habitats, herbivorous insects in the temperate zone usually enter diapause that facilitates synchronization of their life cycle with specific stages of host plants, such as fruit ripening. In the present study, we address those factors regulating dormancy responses as part of a ‘longer strategy’ to persist and thrive in temperate environments, focusing on Rhagoletis cerasi, a univoltine, oligophagous species, which overwinters as pupae and emerges when host fruits are available for oviposition at local scale. To ensure population survival and reproduction at habitats with ecological heterogeneity, R. cerasi has evolved a sophisticated diapause strategy based on a combination of local adaptation and diversified bet‐hedging strategies. Diapause duration is determined both by (i) the adaptive response to local host fruit phenology patterns (annual diapause) and (ii) the plastic responses to unpredictable inter‐annual (temporal) climatic variability that drives a proportion of the populations to extend dormancy by entering a second, successive, facultative cycle of prolonged diapause as part of a bet‐hedging strategy. Besides the dormant periods, post‐diapause development (which varies among populations) exerts ‘fine tune’ adjustments that assure synchronization and may correct possible errors. Adults emerging from pupae with prolonged diapause are larger in body size compared with counterparts emerging during the first year of diapause. However, female fecundity rates are reduced, followed by an extended post‐oviposition period, whereas adult longevity remains unaffected. Overall, it appears that R. cerasi populations are adapted to ecological conditions of local habitats and respond plastically to unpredictable environmental (climatic) conditions.     

PCR-RFLP identification of endemic Chilean species of Rhagoletis (Diptera: Tephritidae) attacking Solanaceae

Four species of the genus Rhagoletis are native to Chile: R. nova (Schiner), R. conversa, (Brèthes), R. penela Foote and R. tomatis Foote. Currently, identification of these species is based on morphological criteria, but their strong similarity makes precise recognition difficult. To clarify species separation for quarantine purposes, a reliable method based on a PCR-RFLP procedure is reported. A DNA region containing mitochondrial NADH dehydrogenase genes was selected as a target sequence for the analysis. The amplification products (c. 1 kb) were digested with either SspI or DdeI, yielding specific patterns that differentiated each of the endemic species. Complete nucleotide sequences were determined, confirming empirical restriction maps. This report updates information on the geographical distribution of Rhagoletis species in Chile.     

Effectiveness of thiamethoxam-coated spheres against blueberry maggot flies (Diptera: Tephritidae)

Studies were conducted to evaluate the mortality of blueberry maggot, Rhagoletis mendax Curran, flies exposed to thiamethoxam- and imidacloprid-coated biodegradable (8-cm-diameter) red spheres, under both laboratory and field conditions. Laboratory studies with spheres coated with 0.1-2.0% (AI) of thiamethoxam indicated that they are effective against R. mendax; however, no dose-dependent response was observed. Studies on the effect of visitation time on thiamethoxam-coated spheres showed a decrease in R. mendax mortality as the duration of visitation time decreased from 60 to 10 s. Under field conditions, significantly more flies were captured on Plexiglas panes below the 2% (AI) thiamethoxam-coated spheres when compared with similar panes below untreated spheres. In field evaluations of thiamethoxam- and imidacloprid-coated spheres, imidacloprid-coated spheres (2.0% [AI]) were found to be significantly more effective than thiamethoxam-coated spheres (0.5-4.0% [AI]). Field trials to characterize the levels of mortality associated with aging pesticide-coated spheres revealed that the effectiveness of treated spheres decreased with increasing age of sphere, and this reduction in effectiveness is greater in thiamethoxam-coated spheres than in imidacloprid-coated spheres. These results provide comparative data on the effectiveness of thiamethoxam- and imidacloprid-coated spheres and support the potential of using pesticide-treated spheres for control of blueberry maggot flies.     

Effect of emergence time on the reproductive parameters of mass-reared Anastrepha ludens (Diptera: Tephritidae) flies

In the mass rearing of fruit flies, the adult emergence time is gradual, due to factors such as variation in food consumption in the larval phase, the female oviposition behavior and environmental conditions, among others. Here, we investigate the effect of emergence time on sex ratio, fecundity, fertility, body size and mating performance in mass-reared Anastrepha ludens (Diptera: Tephritidae) flies. The results of the study indicated that the emergence of the flies is gradual and lasts approximately 96?h, but the highest proportion of emerged flies was observed at 48?h. The flies emerging at 24 and 48?h showed higher fecundity than those emerging at 72 and 96?h, but there were no significant differences in the sex ratio, fertility and male mating performance. Body size was significant, the flies that emerged at 24, 48 and 72?h were bigger than those emerged at 96?h. The discussion focuses on the effect of emergence time on the rearing processes and release of the Mexican fruit fly.     

Distribution,host plant affiliation,phenology, and phylogeny of walnut‐infesting Rhagoletis flies (Diptera: Tephritidae) in Mexico

Many taxa of Nearctic origin have diversified in the subtropical highlands of Mexico. In particular, flies in the genus Rhagoletis have undergone episodes of isolation and gene flow during Pleistocene glaciations and post‐glacial times that have produced lineage differentiation and reproductive isolation. To reach a better understanding of the phylogeography of the genus Rhagoletis, a host plant survey of the walnut‐infesting Rhagoletis suavis species group was conducted across sixteen states comprising 34 different collecting sites in Mexico over a 9‐year period. Five species of Juglans were found to be infested by three species of walnut‐attacking Rhagoletis flies. Several species of parasitoids were also recovered from collections, but in contrast to their walnut fly hosts, they revealed little evidence for host or geographic subdivision. There was no consistent difference in mean eclosion time between walnut fly species or populations associated with different host walnuts in Mexico, unlike the case for other Rhagoletis species, in which allochronic isolation arising from variation in diapause timing is a major ecological adaptation, reproductively isolating flies. We compare the distribution of R. suavis flies in Mexico with those of other Rhagoletis species attacking hawthorns and cherries, and discuss its implications for population divergence and speciation. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2013, 110 , 765–779.     

Evidence for spatial clines and mixed geographic modes of speciation for North American cherry‐infesting Rhagoletis (Diptera: Tephritidae) flies

An important criterion for understanding speciation is the geographic context of population divergence. Three major modes of allopatric, parapatric, and sympatric speciation define the extent of spatial overlap and gene flow between diverging populations. However, mixed modes of speciation are also possible, whereby populations experience periods of allopatry, parapatry, and/or sympatry at different times as they diverge. Here, we report clinal patterns of variation for 21 nuclear‐encoded microsatellites and a wing spot phenotype for cherry‐infesting Rhagoletis (Diptera: Tephritidae) across North America consistent with these flies having initially diverged in parapatry followed by a period of allopatric differentiation in the early Holocene. However, mitochondrial DNA (mtDNA) displays a different pattern; cherry flies at the ends of the clines in the eastern USA and Pacific Northwest share identical haplotypes, while centrally located populations in the southwestern USA and Mexico possess a different haplotype. We hypothesize that the mitochondrial difference could be due to lineage sorting but more likely reflects a selective sweep of a favorable mtDNA variant or the spread of an endosymbiont. The estimated divergence time for mtDNA suggests possible past allopatry, secondary contact, and subsequent isolation between USA and Mexican fly populations initiated before the Wisconsin glaciation. Thus, the current genetics of cherry flies may involve different mixed modes of divergence occurring in different portions of the fly''s range. We discuss the need for additional DNA sequencing and quantification of prezygotic and postzygotic reproductive isolation to verify the multiple mixed‐mode hypothesis for cherry flies and draw parallels from other systems to assess the generality that speciation may commonly involve complex biogeographies of varying combinations of allopatric, parapatric, and sympatric divergence.     

Green fluorescent protein (GFP)-labeling of enterobacteria associated with fruit flies (Diptera: Tephritidae) and persistence in their natural host Rhagoletis completa Cresson

Fruit flies (Diptera: Tephritidae) are a highly successful, widespread group of insects that cause economic damage in agriculture. Data available so far on the composition of the bacterial community associated with their digestive tract indicate that members of Enterobacteriaceae are the species most often isolated. Bacteria naturally occurring in insect guts may be engineered and used to study the spatial and functional interactions of microbes within the insect system and offer one route to meet the demand for novel insect pest management strategies. With this aim we introduced by conjugation the gfp gene carried by the suicide plasmid pTn5gfpmut1 into Klebsiella oxytoca and Raoultella (formerly Klebsiella ) spp. strains isolated from the oesophageal bulb of the fruit flies Ceratitis capitata (Wiedemann) and Rhagoletis completa Cresson, respectively. The GFP-encoding gene was stably maintained in two tested transgenic strains, both originally isolated from R. completa. In one case, GFP-labeled bacterial cells were used to feed larvae and adults of the original host. Genetically modified bacteria were able to colonize the gut of larvae and persisted through all larval instars to pupal stage.