Wolbachia in Anastrepha Fruit Flies (Diptera: Tephritidae)

Endosymbiotic bacteria of the genus Wolbachia are widespread among arthropods and cause a variety of reproductive abnormalities, such as cytoplasmic incompatibility, thelytokous parthenogenesis, male-killing, and host feminization. In this study, we used three sets of Wolbachia-specific primers (16S rDNA, ftsZ, and wsp) in conjunction with the polymerase chain reaction (PCR), cloning and sequencing to study the infection of fruit flies (Anastrepha spp. and Ceratitis capitata) by Wolbachia. The flies were collected at several localities in Brazil and at Guayaquil, Ecuador. All of the fruit flies studied were infected with Wolbachia supergroup A, in agreement with the high prevalence of this group in South America. Phylogenetic analysis showed that the wsp gene was the most sensitive gene for studying the relationships among Wolbachia strains. The Wolbachia sequences detected in these fruit flies were similar to those such as wMel reported for other fruit flies. These results show that the infection of Anastrepha fruit flies by Wolbachia is much more widespread than previously thought.     

Barcoding Queensland Fruit Flies (Bactrocera tryoni): impediments and improvements

Identification of adult fruit flies primarily involves microscopic examination of diagnostic morphological characters, while immature stages, such as larvae, can be more problematic. One of the Australia’s most serious horticultural pests, the Queensland Fruit Fly (Bactrocera tryoni: Tephritidae), is of particular biosecurity/quarantine concern as the immature life stages occur within food produce and can be difficult to identify using morphological characteristics. DNA barcoding of the mitochondrial Cytochrome Oxidase I (COI) gene could be employed to increase the accuracy of fruit fly species identifications. In our study, we tested the utility of standard DNA barcoding techniques and found them to be problematic for Queensland Fruit Flies, which (i) possess a nuclear copy (a numt pseudogene) of the barcoding region of COI that can be co‐amplified; and (ii) as in previous COI phylogenetic analyses closely related B. tryoni complex species appear polyphyletic. We found that the presence of a large deletion in the numt copy of COI allowed an alternative primer to be designed to only amplify the mitochondrial COI locus in tephritid fruit flies. Comparisons of alternative commonly utilized mitochondrial genes, Cytochrome Oxidase II and Cytochrome b, revealed a similar level of variation to COI; however, COI is the most informative for DNA barcoding, given the large number of sequences from other tephritid fruit fly species available for comparison. Adopting DNA barcoding for the identification of problematic fly specimens provides a powerful tool to distinguish serious quarantine fruit fly pests (Tephritidae) from endemic fly species of lesser concern.     

Fruit Flies in Biomedical Research

Many scientists complain that the current funding situation is dire. Indeed, there has been an overall decline in support in funding for research from the National Institutes of Health and the National Science Foundation. Within the Drosophila field, some of us question how long this funding crunch will last as it demotivates principal investigators and perhaps more importantly affects the long-term career choice of many young scientists. Yet numerous very interesting biological processes and avenues remain to be investigated in Drosophila, and probing questions can be answered fast and efficiently in flies to reveal new biological phenomena. Moreover, Drosophila is an excellent model organism for studies that have translational impact for genetic disease and for other medical implications such as vector-borne illnesses. We would like to promote a better collaboration between Drosophila geneticists/biologists and human geneticists/bioinformaticians/clinicians, as it would benefit both fields and significantly impact the research on human diseases.     

Management of Fruit Flies (Diptera: Tephritidae) of the Most Perishable Fruits

We investigated to minimize the dependency on the use of chemicals and thus develop safe and environmental friendly control program for the most perishable fruits i.e., apple,‘ber’, guava and mango. Our findings on the composition of fruit fly species reveal that Bactrocera dorsalis was dominant on apple (33.96% existence), Corpomya incompleta on‘ber’(51.91% existence) and Bactrocera zonata on guava (49.62% existence) and mango (74.66% existence). The correlation between population and infestation percentage was non‐significant in apple orchards, whereas positive and highly significant in between population and infestation, as well as on the cumulative basis in‘ber’, guava and mango orchards during 1998‐1999. Hoeing, baiting and methyl eugenol were statistically equal resulting about 77% decrease in infestation. The maximum control of 91.68% was observed where all four‐control operations including Dipterex® were integrated together. Weather factors, when computed together, had maximum effect on population fluctuation and infestation with rainfall contributing the major role. For guava fruits, the months of August (14.06A individuals/trap/day) and September (13.81A individuals/trap/day) were important, resulting in maximum infestation percentage of 10.76 to 14.74%, respectively.     

湖南省几种实蝇的形态鉴别

综合比较了6种湖南省内捕获的实蝇类昆虫雄成虫的形态鉴别特征,其中离腹寡毛实蝇属(Bactrocera)5种(包括具条实蝇B.scutellata(Hendel)、南瓜实蝇B.tau(Walker)、瓜实蝇B.cucturbitae(Coquillett)、桔小实蝇B.dorsalis(Hendel)和柑橘大实蝇B.(Tetradacus)minax(Enderlein)),以及合腹寡毛实蝇属(Dacus)1种(棍腹实蝇Dacus sp.),以期为实蝇类害虫的检疫和防控提供依据.     

香蕉果实成熟相关基因研究进展(综述)

本文从乙烯生物合成、呼吸作用、碳水化合物代谢、细胞壁降解及其它有关成熟的代谢过程等方面,概述与香蕉果实成熟相关的基因研究进展。     

Experiments on Banana Fruit Development

A number of synthetic auxins and auxin-antagonists were appliedin lanolin emulsions to young banana fruits and their effectson fruit development and seed fertility estimated. Auxins stimulatedthe formation of edible pulp in both seedy and parthenocarpicbananas and induced varying degrees of seed sterility. Of theauxin antagonists tried, coumarin (contrary to a mistaken earlierreport) stimulated parthenocarpy and induced seed sterility,three others were not or but slightly active, and thiourea markedlyinhibited parthenocarpy. Study of the relation between sizeof mature fruit and seed content indicates that developing seedsstimulate the growth of pulp and that the amount of stimulationis roughly proportional to the level of parthenocarpy. Fruitdevelopment in the banana seems to fit in with Luckwill's generalscheme of dual hormonal stimulus.     

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.     

Contrasting Brood-Sex Ratio Flexibility in Two Opiine (Hymenoptera: Braconidae) Parasitoids of Tephritid (Diptera) Fruit Flies

Mass-rearing of fruit fly parasitoids for augmentative release would be more economical if production could be biased towards females. If sex ratios are ever to be manipulated under rearing conditions it is important to determine if, then understand why, sex ratio flexibility exists. Unequal brood-sex ratios are common in parasitic Hymenoptera and two reasons are generally advanced for such cases: 1) avoidance of Local Mate Competition (LMC) predicts that the number of females exploiting a host-patch can influence the optimal sex ratio of their offspring; and 2) one sex is more likely to develop under a particular set of physical/competitive conditions. We hypothesized that LMC is more often encountered in relatively uncommon species with coarse-grained distributions. As a result, isolated females of such species would be more likely to expect future LMC to be high and to bias offspring sex ratios towards females. We proposed that the opiine braconid Utetes anastrephae is such a coarse-grained species and compared its responses to differences to LMC (isolated females and those paired with a conspecific competitor) with those of another opiine, Diachasmimorpha longicaudata believed to have a finer-grained distribution. Adult sex ratios were mutable in D. longicaudata and U. anastrephae and, as predicted by avoidance of LMC, males were relatively more abundant among the paired females in both species. However, superparasitized hosts yielded relatively more daughters, perhaps because female larvae are superior intrinsic competitors. Contrary to prediction, there was no suggestion that U. anastrephae was more likely to than D. longicaudata to avoid LMC. While these results did not reveal any species differences in sex-allocation patterns or suggest any modifications to the present practice of fruit fly mass-rearing for augmentative release they did provide new information regarding U. anastrephae, a widespread natural enemy of Anastrepha spp. about which relatively little is known.     

槟榔烂果病的病原菌鉴定

本研究分别从台湾槟榔病果中分离出病原菌Ar8904、Ar8905;从海南槟榔病果中分离出病原菌Ar8801、Ar8802、Ar8904以及从云南槟榔病果中分离出病原菌Ar8901、Ar892.将这些病原菌进行对比实验.研究结果表明,这7株病原菌在菌株形态、培养形状、寄主范围、致病性以及症状特征都十分的相似.因此,根据炭疽菌现代分类标准,我们可以初步确认台湾、海南、云南的槟榔病果均为同一种炭疽菌引起的.     

The Development of the Banana Fruit

1. The banana fruit is either seeded or vegetatively parthenocarpic;the latter may or may not be seed fertile, depending on a complexof cytogenetical factors. Edible pulp (a starchy parenchyma),which fills the fruit in parthenocarpic types and surroundsthe seeds in seeded bananas, mostly originates from the outerlining of the loculus (the innermost layer of the pericarp). 2. The graphs of growth in volume of seeded banana fruits aresigmoid in shape. Those of parthenocarpic fruits are variablebut are not sigmoid and the shapes are related to specific origins. 3. Growth rates are related to certain ovule behaviours, toseed content of the fruit, and to ploidy. 4. -Naphthylacetic acid induces parthenocarpy in seeded bananasand stimulates it in weakly parthenocarpic types. By contrast,coumarin, a hormone inhibitor, inhibits it in strongly parthenocarpicforms. 5. Physiological and genetical implications of the results arediscussed. Fruit development is thought to be under hormonecontrol and two phases of development of the parthenocarpicfruit can be distinguished. The desirability of distinguishingbetween the terms ‘seedless’ and ‘parthenocarpic’is pointed out.     

The Value of Patch-Choice Copying in Fruit Flies

Many animals copy the choices of others but the functional and mechanistic explanations for copying are still not fully resolved. We relied on novel behavioral protocols to quantify the value of patch-choice copying in fruit flies. In a titration experiment, we quantified how much nutritional value females were willing to trade for laying eggs on patches already occupied by larvae (social patches). Females were highly sensitive to nutritional quality, which was positively associated with their offspring success. Females, however, perceived social, low-nutrition patches (33% of the nutrients) as equally valuable as non-social, high-nutrition ones (100% of the nutrients). In follow-up experiments, we could not, however, either find informational benefits from copying others or detect what females'' offspring may gain from developing with older larvae. Because patch-choice copying in fruit flies is a robust phenomenon in spite of potential costs due to competition, we suggest that it is beneficial in natural settings, where fruit flies encounter complex dynamics of microbial communities, which include, in addition to the preferred yeast species they feed on, numerous harmful fungi and bacteria. We suggest that microbial ecology underlies many cases of copying in nature.     

Lek Site Selection by Male Mediterranean Fruit Flies

The mating system of the Mediterranean fruit fly, Ceratitis capitata (Wiedemann), is based on leks. In field cage choice experiments we examined lek site selection as influenced by the presence of fruit and the presence of conspecific flies. Males were allowed to choose between artificial trees containing fruit and trees without fruit. Males showed a significant preference for the tree baited with fruit. To determine which fruit-related stimuli were important, males were allowed to choose between visual and/or olfactory fruit-derived stimuli. Males preferred to lek on trees presenting both stimuli. To determine the influence of conspecifics on lek site selection, males were allowed to choose between trees containing male and/or female conspecific flies. The presence of pheromone-emitting males was preferred. In addition, both in the field and on artificial trees, we examined the effect of leaf size on lek site selection. In the field, males preferred to lek on leaves of a particular size. Moreover, leaf integrity was important, as males tended to select undamaged leaves as lek sites. In field cage experiments, males consistently chose to lek on trees containing the largest leaves. We conclude that the choice of lek site is influenced by the presence of fruit and of other lekking males, while leaf size and integrity determine male location within a lek.     

Kinetics of Colonization of Adult Queensland Fruit Flies (Bactrocera tryoni) by Dinitrogen-Fixing Alimentary Tract Bacteria

The average total population of bacteria remained constant in the alimentary tracts of adult laboratory-raised Queensland fruit flies (Bactrocera tryoni) although the insects had ingested large numbers of live bacteria as part of their diet. The mean number of bacteria (about 13 million) present in the gut of the insects from 12 to 55 days after emergence was not significantly modified when, at 5 days after emergence, the flies were fed antibiotic-resistant bacteria belonging to two species commonly isolated from the gut of field-collected B. tryoni. Flies were fed one marked dinitrogen-fixing strain each of either Klebsiella oxytoca or Enterobacter cloacae, and the gastrointestinal tracts of fed flies were shown to be colonized within 7 days by antibiotic-resistant isolates of K. oxytoca but not E. cloacae. The composition of the microbial population also appeared to be stable in that the distribution and frequency of bacterial taxa among individual flies exhibited similar patterns whether or not the flies had been bacteria fed. Isolates of either E. cloacae or K. oxytoca, constituting 70% of the total numbers, were usually dominant, with oxidative species including pseudomonads forming the balance of the population. Antibiotic-resistant bacteria could be spread from one cage of flies to the adjacent surfaces of a second cage within a few days and had reached a control group several meters distant by 3 weeks. Restriction of marked bacteria to the population of one in five flies sampled from the control group over the next 30 days suggested that the bacterial population in the gut of the insect was susceptible to alteration in the first week after emergence but that thereafter it entered a steady state and was less likely to be perturbed by the introduction of newly encountered strains. All populations sampled, including controls, included at least one isolate of the dinitrogen-fixing family Enterobacteriaceae; many were distinct from the marked strains fed to the flies. Nitrogenase activity detected by the acetylene reduction assay was associated with flies fed dinitrogen-fixing bacteria as well as with control groups given either no supplement or free access to a yeast hydrolysate preparation. Nitrogen fixed from the atmosphere may supplement the nutrition of the alimentary tract microbial population of B. tryoni. Transmission electron microscopy showed that the principal site of bacterial colonization in the abdominal alimentary tract was the lumen of the midgut inside the peritrophic membrane. No intracellular symbionts were seen in the gut tissues nor were bacteria found attached to the cuticular folds of the hindgut. The ultrastructure of the gut resembled that of other fly genera except that the intercellular spaces between rectal epithelial cells were more extensive, suggesting a role for unspecialized epithelium in water and solute uptake in B. tryoni.     

The Bionomics ofCoptera Haywardi(Ogloblin) (Hymenoptera: Diapriidae) and Other Pupal Parasitoids of Tephritid Fruit Flies (Diptera)

The endoparasitoidCoptera haywardi(Ogloblin) (Diapriidae) was discovered in Mexico attacking the pupae of the Mexican fruit fly,Anastrepha ludens(Loew). Typically, parasitoids of Diptera Cychlorrhapha pupae develop as ectoparasitoids and are generalists that attack hosts in a number of families. Aspects of the bionomics ofC. haywardiwere compared to those of two chalcidoid ectoparasitoids,Dirhinus himalayanusWestwood andSpalangia geminaBoucek.C. haywardideveloped in three genera of Tephritidae, but not in species of other families. The two species of chalcidoids developed in all the calypterate and acalypterate hosts to which they were exposed. In an olfactometerC. haywardipreferredAnastrepha suspensa(Loew) pupae, while the chalcidoids preferred the pupae ofMusca domestica L.This preference inS. geminawas diminished in insects that had been reared onA. suspensa. C. haywardioviposited in theA. suspensapupae that had been previously parasitized by the braconidDiachasmimorpha longicaudata(Ashmead). However, it completed development only in unparasitized pupae. Mortality of the primary parasitoid due toD. himalayanuswas approximately two-thirds the mortality inflicted on the host fly.S. geminadid not discriminate between parasitized and unparasitized pupae ofA. suspensaand developed in both.C. haywardiappears to have a more restricted host range relative to chalcidoid pupal parasitoids and this may be due to its endoparasitic development.     

Characterization and Evolution of mariner Elements from Closely Related Species of Fruit Flies (Diptera: Tephritidae)

Mariner elements were amplified using the polymerase chain reaction from two species of tephritid flies, Ceratitis rosa and Trirhithrum coffeae. The sequences were ∼1.3 kb in length. None of these elements appeared to be functional, as in every case the open reading frame (ORF) was disrupted by the presence of frameshifts or stop codons. These elements, Crmar1 and Tcmar1, are very similar to the Ccmar1 element previously amplified from the closely related tephritid species C. capitata and are members of the mellifera subfamily of mariner elements. The phylogeny and pattern of divergence of these elements were examined in relation to the phylogeny of the host species. It is highly probable that the elements were present in the ancestral lineage prior to the divergence of the three species. The copy numbers of the elements within each species are very different, ranging from about 10 in T. coffeae to 5,000 in C. rosa. The possible mechanisms which determine the copy number of an element in the host genome are discussed. Received: 25 April 1997 / Accepted: 31 July 1997