Characteristics of protein variants in trichlorphon-resistant Bactrocera dorsalis (Diptera; Tephritidae) larvae

Functional proteins in larvae of Bactrocera dorsalis, a major fruit pest, play a central role in their resistance to organophosphorus insecticides. Changes in proteins in B. dorsalis larvae after trichlorphon treatment may have a role in the resistance response to trichlorphon. We analyzed 14 protein spots of crude proteins from B. dorsalis larvae post-treatment with trichlorphon in two-dimensional gel electrophoresis through mass spectrometry and protein sequencing. We found functional proteins that are responsible for signal transduction (pkaap and dual specificity tyrosine-phosphorylation-regulated kinase), immunity (hemolectin), synthesis and decomposition (twinstar, cathepsin B, RE66325p), oxidative stress metabolism (glutathione S transferase and CG7320), energy metabolism (Act57B), and cytoskeleton formation (actin). These proteins appear to be involved in the resistance response to trichlorphon.     

Resistance and synergistic effects of insecticides in Bactrocera dorsalis (Diptera: Tephritidae) in Taiwan

Oriental fruit flies, Bactrocera dorsalis (Hendel), were treated with 10 insecticides, including six organophosphates (naled, trichlorfon, fenitrothion, fenthion, formothion, and malathion), one carbamate (methomyl), and three pyrethroids (cyfluthrin, cypermethrin, and fenvalerate), by a topical application assay under laboratory conditions. Subparental lines of each generation treated with the same insecticide were selected for 30 generations and were designated as x-r lines (x, insecticide; r, resistant). The parent colony was maintained as the susceptible colony. The line treated with naled exhibited the lowest increase in resistance (4.7-fold), whereas the line treated with formothion exhibited the highest increase in resistance (up to 594-fold) compared with the susceptible colony. Synergism bioassays also were carried out. Based on this, S,S,S-tributyl phosphorotrithioate displayed a synergistic effect for naled, trichlorfon, and malathion resistance, whereas piperonyl butoxide displayed a synergistic effect for pyrethroid resistance. All 10 resistant lines also exhibited some cross-resistance to other insecticides, not only to the same chemical class of insecticides but also to other classes. However, none of the organophosphate-resistant or the methomyl-resistant lines exhibited cross-resistance to two of the pyrethroids (cypermethrin and fenvalerate). Overall, the laboratory resistance and cross-resistance data developed here should provide useful tools and information for designing an insecticide management strategy for controlling this fruit fly in the field.     

Sublethal and transgenerational effects of six insecticides on Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

To clarify the effects of sublethal concentration of insecticide on the growth, development and reproduction of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), the sublethal and transgenerational effects of the LC₃₀ of six insecticides on adults and progeny (F₁ and F₂ generations) were studied using age-stage, two-sex life tables. Exposure to six insecticides had significant negative impacts on the life-history traits of B. dorsalis and led to reduced adult longevity and fecundity in F₀, a longer larval stage duration in F₁, and reduced fertility and survival. The intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rate (R₀) were reduced in F₁, with the greatest reduction observed in the chlorpyrifos treatment. In the F₂ generation, the adverse effects were weakened, only the cyantraniliprole treatment had lower values than the control, and the fecundity of other treatments returned to normal. Obvious “hormesis” phenomena were not observed in any of the generations. Based on the population hazard index, chlorpyrifos had the best control effect in the early stage, and cyantraniliprole had the best control effect in the long term. These results indicate that the LC₃₀ of the six insecticides exert different transgenerational effects on the B. dorsalis population.     

桔小实蝇及其寄生性天敌——前裂长管茧蜂饲养方法

系统介绍了桔小实蝇Bactrocera dorsalis（Hendel）室内饲养的设备、步骤、技术要点、人工饲料配方及环境要求,并以桔小实蝇为寄主,实现了桔小实蝇幼虫一蛹期寄生性天敌前裂长管茧蜂Diachasmimorpha longicaudata（Ashmead）的室内大量繁殖,为进一步利用该蜂防治桔小实蝇打下了良好的基础。     

橘小实蝇交配行为观察

在实验室内对橘小实蝇Bactrocera dorsalis（Hendel）的交配行为进行了观察。描述了橘小实蝇交配中雄虫的“求偶场”,雄雄相遇、雌雌相遇、雌雄相遇时的相互反应,雌雄虫的交配行为与交配后行为。     

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.     

Odorant receptor co-receptor Orco is upregulated by methyl eugenol in male Bactrocera dorsalis (Diptera: Tephritidae)

Bactrocera dorsalis is a destructive fruit-eating pest that causes severe economic damage to the fruit and vegetable industry. Methyl eugenol (ME) has been widely used as an effective sexual attractant for male fruit flies through olfactory perception. However, the molecular mechanism underlying the olfactory perception of ME remains unknown. Here, we report the characterization and functional analysis of a newly discovered cDNA that encodes a Drosophila melanogaster odorant receptor co-receptor Orco ortholog in B. dorsalis. qRT-PCR analysis revealed that it was abundantly expressed in the antenna of adult B. dorsalis. Notably, Orco was upregulated by ME in the antenna of male flies. Mature males of B. dorsalis showed significant taxis toward ME within 0.5h, and Orco was significantly upregulated in the attracted adults within the same period. Silencing Orco through the ingestion of dsRNA reduced the attractive effects of ME. These data suggest that Orco may play an essential role in ME attraction in the olfactory signal transduction pathway.     

Response of Ceratitis capitata, Bactrocera dorsalis, and Bactrocera cucurbitae (Diptera: Tephritidae) to metabolic stress disinfection and disinfestation treatment

Metabolic stress disinfection and disinfestation (MSDD) is a postharvest treatment designed to control pathogens and arthropod pests on commodities that combines short cycles of low pressure/vacuum and high CO2 with ethanol vapor. Experiments were conducted to evaluate the effect of MSDD treatment on various life stages of Ceratitis capitata (Wiedemann), Mediterranean fruit fly; Bactrocera dorsalis Hendel, oriental fruit fly; and Bactrocera cucurbitae Coquillett, melon fly, in petri dishes and in papaya, Carica papaya L., fruit. In some experiments, the ethanol vapor phase was withheld to separate the effects of the physical (low pressure/ambient pressure cycles) and chemical (ethanol vapor plus low pressure) phases of treatment. In the experiments with tephritid fruit fly larvae and adults in petri dishes, mortality was generally high when insects were exposed to ethanol and low when ethanol was withheld during MSDD treatment, suggesting that ethanol vapor is highly lethal but that fruit flies are quite tolerant of short periods of low pressure treatment alone. When papaya fruit infested with fruit fly eggs or larvae were treated by MSDD, they produced fewer pupae than untreated control fruit, but a substantial number of individuals developed nonetheless. This suggests that internally feeding insects in fruit may be partially protected from the toxic effects of the ethanol because the vapor does not easily penetrate the fruit pericarp and pulp. MSDD treatment using the atmospheric conditions tested has limited potential as a disinfestation treatment for internal-feeding quarantine pests such as fruit flies infesting perishable commodities.     

Mitotic and polytene chromosomes analysis of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

The Oriental fruit fly, Batrocera dorsalis s.s. (Hendel) is one of the most destructive agricultural pests, belonging to a large group of difficult to distinguish morphologically species, referred as the B. dorsalis complex. We report here a cytogenetic analysis of two laboratory strains of the species and provide a photographic polytene chromosome map from larval salivary glands. The mitotic complement consists of six chromosome pairs including a heteromorphic sex (XX/XY) chromosome pair. Analysis of the polytene complement has shown a total of five polytene chromosomes (10 polytene arms) that correspond to the five autosomes. The most important landmarks of each polytene chromosome and characteristic asynapsis at a specific chromosomal region are presented and discussed. Chromosomal homology between B. dorsalis and Ceratitis capitata has been determined by comparing chromosome banding patterns. The detection of chromosome inversions in both B. dorsalis strains is shown and discussed. Our results show that the polytene maps presented here are suitable for cytogenetic analysis of this species and can be used for comparative studies among species of the Tephritidae family. They also provide a diagnostic tool that could accelerate species identification within the B. dorsalis complex and could shed light on the ongoing speciation in this complex. Polytene chromosome maps can facilitate the development of biological control methods and support the genome mapping project of the species that is currently in progress.     

中国果实蝇属一新种记述(双翅目:实蝇科:寡鬃实蝇亚科)

记述自中国云南的果实蝇属Bactrocera Macquart 1新种,即保山果实蝇Bactrocera (Zeugodacus)baoshanensis,sp.nov..模式标本保存于福建农林大学益虫研究所.     

Cold treatment for guava fruits infested with oriental fruit fly,Bactrocera dorsalis (Diptera: Tephritidae)

Guava (Psidium guajava Linnaeus) is an important horticulture crop in tropical and subtropical regions. However, guava transportation to the fruit market is regulated due to the risk of fruit fly infestations. The oriental fruit fly [Bactrocera dorsalis (Hendel), Tephritidae, Diptera], which damages numerous horticulture fruits, is the primary pest in guava. For fruit export, phytosanitary treatment is necessary to prevent the potential oriental fruit fly infestations through pest-infested fruits. In this study, cold treatment was administered to eliminate oriental fruit fly infestations in guava fruits. The optimal egg inoculation density was 200 eggs per fruit, and third instar larvae displayed the highest cold treatment tolerance. On assessing the efficacy of cold treatment, none of the third instar larvae in the fruits survived at a constant fruit core temperature of 0.5–1 °C for 11 days. A confirmatory test revealed that on maintaining a fruit core temperature of 0.5–1 °C for 12 days, 100% mortality was achieved among the fruit flies. In fruit quality assessment, guava fruits exposed to cold treatment still maintained their market value. Thus, cold treatment effectively disinfested guava fruits with possible fruit fly infestations, making it a viable quarantine treatment for guava fruits prior to their export.     

The current and future potential geographical distribution of the oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae)

The oriental fruit fly, Bactrocera dorsalis (Hendel), is a major pest throughout South East Asia and in a number of Pacific Islands. As a result of their widespread distribution, pest status, invasive ability and potential impact on market access, B. dorsalis and many other fruit fly species are considered major threats to many countries. CLIMEX was used to model the potential global distribution of B. dorsalis under current and future climate scenarios. Under current climatic conditions, its projected potential distribution includes much of the tropics and subtropics and extends into warm temperate areas such as southern Mediterranean Europe. The model projects optimal climatic conditions for B. dorsalis in the south-eastern USA, where the principle range-limiting factor is likely to be cold stress. As a result of climate change, the potential global range for B. dorsalis is projected to extend further polewards as cold stress boundaries recede. However, the potential range contracts in areas where precipitation is projected to decrease substantially. The significant increases in the potential distribution of B. dorsalis projected under the climate change scenarios suggest that the World Trade Organization should allow biosecurity authorities to consider the effects of climate change when undertaking pest risk assessments. One of the most significant areas of uncertainty in climate change concerns the greenhouse gas emissions scenarios. Results are provided that span the range of standard Intergovernmental Panel on Climate Change scenarios. The impact on the projected distribution of B. dorsalis is striking, but affects the relative abundance of the fly within the total suitable range more than the total area of climatically suitable habitat.     

Biology of Bactrocera carambolae (Diptera: Tephritidae) on four hosts

Bactrocera carambolae is a quarantine pest found in Brazil, restricted to the states of Amapá, Pará and Roraima. This fruit fly can potentially cause extensive socioeconomic and environmental damage in the country, if it disperse into areas where fruit is grown for exporting. The objective of this work was to study the biology of B. carambolae on fruits of Averrhoa carambola L. (Oxalidaceae), Psidium guajava L. (Myrtaceae), Spondias mombin L. (Anacardiaceae) and Eugenia stipitata McVaugh (Myrtaceae). The following parameters were investigated: duration of egg-larva, pupal, egg-adult, pre-oviposition, oviposition and post-oviposition periods, pupal weight and viability, sex ratio, fecundity, fertility and longevity. All parameters except pupal weight, oviposition and post-oviposition period, egg fertility and sex ratio were influenced by the host plant on which the larvae were reared. The carambola fruit fly completes its development on all those hosts studied here, with the highest fecundities on A. carambola and P. guajava.     

橘小实蝇幼虫液体人工饲料中pH值对其生长发育的影响

本文主要研究了橘小实蝇液体人工幼虫饲料中不同酸度对橘小实蝇生长发育的影响。继代饲养3代的结果表明：饲料酸度对橘小实蝇化蛹率和后代卵孵化率没有影响;对橘小实蝇蛹重、成虫羽化率和产卵量有显著影响。橘小实蝇液体人工幼虫饲料的pH值调节至3.6时仍可以获得质量较高的橘小实蝇。     

The complete nucleotide sequence of the mitochondrial genome of the oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae)

The complete mitochondrial genome of the oriental fruit fly Bactrocera dorsalis s.s. has been sequenced, and is here described and compared with the homologous sequences of Bactrocera oleae and Ceratitis capitata. The genome is a circular molecule of 15,915 bp, and encodes the set of 37 genes generally found in animal mitochondrial genomes. The structure and organization of the molecule is typical and similar to the two closely related species B. oleae and C. capitata, although it presents an interesting case of putative intra-molecular recombination. The relevance of the growing comparative dataset of tephritid complete mitochondrial genomes is discussed in relation to the possibility to develop robust assays for species discrimination in quarantine and agricultural monitoring practices, as well as basic phylogeography/population genetic studies.