Oviposition Site-Selection by Bactrocera dorsalis Is Mediated through an Innate Recognition Template Tuned to γ-Octalactone

Innate recognition templates (IRTs) in insects are developed through many years of evolution. Here we investigated olfactory cues mediating oviposition behavior in the oriental fruit fly, Bactrocera dorsalis, and their role in triggering an IRT for oviposition site recognition. Behavioral assays with electrophysiologically active compounds from a preferred host, mango, revealed that one of the volatiles tested, γ-octalactone, had a powerful effect in eliciting oviposition by gravid B. dorsalis females. Electrophysiological responses were obtained and flies clearly differentiated between treated and untreated substrates over a wide range of concentrations of γ-octalactone. It triggered an innate response in flies, overriding inputs from other modalities required for oviposition site evaluation. A complex blend of mango volatiles not containing γ-octalactone elicited low levels of oviposition, whereas γ-octalactone alone elicited more oviposition response. Naïve flies with different rearing histories showed similar responses to γ-octalactone. Taken together, these results indicate that oviposition site selection in B. dorsalis is mediated through an IRT tuned to γ-octalactone. Our study provides empirical data on a cue underpinning innate behavior and may also find use in control operations against this invasive horticultural pest.     

Is a mango just a mango? Testing within-fruit oviposition site choice and larval performance of a highly polyphagous fruit fly

For fruit flies, fully ripe fruit is preferred for adult oviposition and is superior for offspring performance over unripe or ripening fruit. Because not all parts of a single fruit ripen simultaneously, the opportunity exists for adult fruit flies to selectively choose riper parts of a fruit for oviposition and such selection, if it occurs, could positively influence offspring performance. Such fine scale host variation is rarely considered in fruit fly ecology, however, especially for polyphagous species which are, by definition, considered to be generalist host users. Here we study the adult oviposition preference/larval performance relationship of the Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), a highly polyphagous pest species, at the “within-fruit” level to see if such a host use pattern occurs. We recorded the number of oviposition attempts that female flies made into three fruit portions (top, middle and bottom), and larval behavior and development within different fruit portions for ripening (color change) and fully-ripe mango, Mangifera indica L. (Anacardiaceae). Results indicate that female B. dorsalis do not oviposit uniformly across a mango fruit, but lay most often in the top (i.e., stalk end) of fruit and least in the bottom portion, regardless of ripening stage. There was no evidence of larval feeding site preference or performance (development time, pupal weight, percent pupation) being influenced by fruit portion, within or across the fruit ripening stages. There was, however, a very significant effect on adult emergence rate from pupae, with adult emergence rate from pupae from the bottom of ripening mango being approximately only 50% of the adult emergence rate from the top of ripening fruit, or from both the top and bottom of fully-ripe fruit. Differences in mechanical (firmness) and chemical (total soluble solids, titratable acidity, total non-structural carbohydrates) traits between different fruit portions were correlated with adult fruit utilisation. Our results support a positive adult preference/offspring performance relationship at within-fruit level for B. dorsalis. The fine level of host discrimination exhibited by B. dorsalis is at odds with the general perception that, as a polyphagous herbivore, the fly should show very little discrimination in its host use behavior.     

Effects of Co-60 radiation on the activities of three main antioxidant enzymes in Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

Co-60 radiation has been broadly used for pest management because it results in male sterility and a lack of emergence of unparasitized hosts due to oxidative damage. Insect life is significantly affected by abiotic stress factors, including Co-60 radiation. In the present study, we determined the influence of Co-60 radiation on an important pest in southern China, the oriental fruit fly Bactrocera dorsalis (Diptera: Tephritidae). We exposed 30-h-old B. dorsalis eggs to Co-60 radiation at doses of (0, 5, 10, 15, 20, 25, 30, 35, and 40?Gy) and determined the activity of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and peroxidase (POX). CAT and POX activity significantly increased in B. dorsalis in the 1st larvae instar after high-intensity radiation doses (25–40?Gy) and played an important role in the antioxidant response to intensive radiation. The activity of POX in B. dorsalis at the 2nd larval instar sharply decreased after different doses of radiation, except for 10 and 40?Gy. Our results demonstrated that Co-60 radiation affected the activity of antioxidant enzymes and disturbed the physiology of B. dorsalis, especially at early stages (the 1st, 2nd larvae instar). These three antioxidant defense enzymes cooperatively play an important role in protecting B. dorsalis from oxidative damage.     

Attraction of feral Bactrocera dorsalis males (Diptera: Tephritidae) to natural versus commercial sources of methyl eugenol

Males of Bactrocera dorsalis (Hendel) are strongly attracted to methyl eugenol (ME), which occurs in over 450 plant species. Given this powerful attraction, ME is commonly used in surveillance and eradication programs against this invasive agricultural pest. Preliminary observations revealed that B. dorsalis males visited ME-bearing flowers of the fruit fly orchid (FFO) Bulbophyllum cheiri subsp. cheiri even when these occurred near traps baited with far greater quantities of ME. Based on this evidence, we undertook field experiments to assess the attraction of feral B. dorsalis males to FFO flowers relative to commercial sources of ME. At the edge of a secondary forest, an FFO flower was placed midway between two ME sources located 20 m apart, and attracted flies were collected over an entire day. When the ME sources were unenclosed (not in traps), the relative attractiveness of FFO flowers to B. dorsalis males varied with the amount of ME placed nearby. The FFO flower (i) attracted a similar proportion of males when 1 g ME was placed at the flanking sites but (ii) captured significantly smaller proportions when the nearby sites had 6 or 10 g commercial ME sources. Similar tests with the commercial sources enclosed in traps showed that (i) 6 g ME sources in Steiner traps attracted significantly more B. dorsalis males than FFO flowers but (ii) 10 g sources in Clear traps (1 L buckets with 4 entrance holes) did not outperform the FFO flowers, presumably owing to limited dispersion of volatiles from the trap. Implications of these results for surveillance programs are discussed.     

桔小实蝇和番石榴实蝇对6种寄主果实的产卵选择适应性

植食性昆虫对寄主植物的选择适应性是研究昆虫和植物协同进化关系的核心内容之一。评估寄主植物对植食性昆虫种群的适合度,需要综合分析昆虫对寄主的产卵选择性和寄主对昆虫的取食适合性。以桔小实蝇和番石榴实蝇为研究对象,分别测定了这两种实蝇对6种寄主果实:番石榴、香蕉、杨桃、木瓜、甜橙、番茄的产卵选择性以及幼虫取食后对其生长发育的影响。寄主产卵选择性实验分别采用完整寄主果实直接供试产卵和块状寄主果实气味引诱产卵两种处理方式;在生长发育适应性实验中,以幼虫和蛹的存活和生长发育等相关参数作为评价指标。实验结果表明,寄主的供试方式不同,两种实蝇的选择性均有明显差异;对寄主气味选择性强的寄主更适合于两种实蝇后代的生长发育。两种实蝇对6种寄主果实的产卵选择性和后代发育适合性两者相关性不显著,与许多文献报道单一地采用发育适合性(如发育历期、存活率或蛹重等)作为评价寄主选择性的结果不一致。两种实蝇之间对6种寄主果实的产卵选择和幼虫取食适合性既具相似性也具差异性,表明这两种实蝇在寄主生态位上既有重叠性又有分化性。     

What signals do herbivore-induced plant volatiles provide conspecific herbivores?

Herbivore-induced plant volatiles (HIPVs) have been opined as ‘indirect or direct defenses’ of plants and are extensively studied. In contrast, HIPVs may also indicate that plant defenses have been overcome by herbivores infesting the plant; however, studies on this aspect have so far received little attention. Using the interaction of Capsicum annum (Bell pepper) with its pest Scirtothrips dorsalis (Chilli thrips) as a model system, we studied the role of HIPVs in this selected insect–plant interaction. Multiple-choice olfactometer assays with headspace volatiles collected from different growth stages of un-infested C. annum plants represented by pre-flowering (PF), flowering (FL) and fruiting stages (FR) proved FR volatiles to be highly attractive to S. dorsalis. Further, FR plants were infested with S. dorsalis adults and HIPVs released by infested plants were collected and subjected to multiple-choice olfactometer bioassays. Thrips were significantly attracted to HIPVs than to headspace volatiles of un-infested FR plants or thrips body odour. Coupled GC-EAG with S. dorsalis and HIPVs or FR plant volatile revealed specific compounds that elicited an EAG response. Individual EAG-active compounds were less attractive to thrips, however, synthetic blends of EAG-active compounds at the ratio similar to headspace samples were found to be highly attractive. However, when given a choice between synthetic blends of HIPVs and FR, thrips were significantly attracted to synthetic blend of HIPVs. Our study provides empirical data on signals HIPVs may provide to conspecific herbivores and suggests that the role of HIPVs, mostly generalized as defense, may vary based on the interaction and must be studied closely to understand their ecological functions.     

Suboptimal Larval Habitats Modulate Oviposition of the Malaria Vector Mosquito Anopheles coluzzii

Selection of oviposition sites by gravid females is a critical behavioral step in the reproductive cycle of Anopheles coluzzii, which is one of the principal Afrotropical malaria vector mosquitoes. Several studies suggest this decision is mediated by semiochemicals associated with potential oviposition sites. To better understand the chemosensory basis of this behavior and identify compounds that can modulate oviposition, we examined the generally held hypothesis that suboptimal larval habitats give rise to semiochemicals that negatively influence the oviposition preference of gravid females. Dual-choice bioassays indicated that oviposition sites conditioned in this manner do indeed foster significant and concentration dependent aversive effects on the oviposition site selection of gravid females. Headspace analyses derived from aversive habitats consistently noted the presence of dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS) and 6-methyl-5-hepten-2-one (sulcatone) each of which unitarily affected An. coluzzii oviposition preference. Electrophysiological assays across the antennae, maxillary palp, and labellum of gravid An. coluzzii revealed differential responses to these semiochemicals. Taken together, these findings validate the hypothesis in question and suggest that suboptimal environments for An. coluzzii larval development results in the release of DMDS, DMTS and sulcatone that impact the response valence of gravid females.     

Fruit trap: A detection and collection tool for opiine parasitoids (Hym.: Braconidae) of the oriental fruit fly,Bactrocera dorsalis (Dipt.: Tephritidae)

A fruit trap was developed for detection and collection of the opiine parasitoids of the oriental fruit fly,Bactrocera (=Dacus)dorsalis (Hendel). Gravid females ofBiosteres arisanus (Sonan), an egg-larval parasitoid, orDiachasmimorpha longicaudata (Ashmead) andPsytallia incisi (Silvestri), both larval parasitoids, were lured to parasitize the eggs or larvae ofB. dorsalis inoculated in ripe papaya fruits,Carica papaya L. Progenies ofB. arisanus were consistently recovered from papaya fruits inoculated withB. dorsalis eggs (subsequently referred to as egg fruit traps). Except in Moloaa on Kauai (6%), higher percentage ofB. dorsalis parasitization (range=38–43%) was recorded in Hilo, island of Hawaii and Waimanalo and Poamoho, island of Oahu. Progenies ofD. longicaudata and a fewP. incisi were recovered from papaya fruits artificially infested withB. dorsalis larvae (subsequently referred to as larval fruit traps). The recovery of parasitoid progenies from larval fruit traps suspended from papaya trees did not differ significantly from larval fruit traps placed on the ground. In both methods of trap placement, percent parasitization ofB. dorsalis byD. longicaudata (predominant species) ranged from 58–60%. On the other hand, significantly moreB. arisanus thanD. longicaudata andP. incisi adults (larval parasitoids) were recovered from fully ripened to highly deteriorated papaya fruits collected from papaya trees or ground (fallen fruits).     

γ‐Octalactone,an effective oviposition stimulant of Bactrocera tryoni

Insects commonly rely on olfactory, gustatory and visual cues when deciding where to lay eggs. The olfactory cues that stimulate oviposition in the Queensland fruit fly, Bactrocera tryoni (Froggatt) (Diptera: Tephritidae), are not well understood. Here, we show that two known oviposition stimulants of the Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)—γ‐octalactone and benzothiazole—strongly elicit aggregation and oviposition in B. tryoni. Two other known oviposition stimulants of B. dorsalis—ethyl tiglate and 1‐octen‐3‐ol—elicit aggregation but not oviposition. Highlighting species overlap, but also differences, in oviposition stimulants, these findings have practical application for mass‐rearing in which vast numbers of flies are reared for sterile insect technique programs and may also have practical application in the development of pest management and monitoring tools.     

A tentative evaluation for population establishment of Bactrocera dorsalis (Diptera: Tephritidae) by its population modeling: Considering the temporal distribution of host plants in a selected area in Jeju,Korea

The Oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) is a destructive fruit pest in a wide range of cultivated fruit crops and wild plants. This species is a potentially highly invasive fruit fly to Jeju area of Korea. This study was conducted to evaluate the effect of host plants distributed temporally on the population development of B. dorsalis. The temperature-dependent bionomic data for a synonymous group of B. dorsalis, including B. philippinensis, B. papayae, and B. invadens were collected from previous publications and combined to construct a population model of this pest and its thermal constant. We developed a stage-transition model of eggs, larvae and pupae, and an oviposition model for basic population modeling of the four common strains. We investigated the abundance of the host plants of B. dorsalis in a selected site in Jeju and parameterized them in terms of temporal availability to incorporate into the population model. The contribution of host plants for the population growth of B. dorsalis in the selected site was different according to the group of host plants. For example, B. dorsalis populations largely decreased by 93%, when host plants belong to Moraceae (mainly Ficus sp.) were removed in the simulation. Also, we found that the host plants of Prunus persica, Ficus carica, P. mume, Eriobotrya japonica in this order contributed greatly to population abundance of B. dorsalis in the selected area, which was important in terms of mid-season host plants connecting the early adult population of B. dorsalis to citrus plants in the late season. Finally, we discussed a seasonal management strategy against B. dorsalis while considering the availability of host plants and the biology of this fruit fly in an invaded area.     

Volatiles from host plant brinjal attract the brinjal Fruit and Shoot Borer -Leucinodes orbonalis Guenee

Brinjal Fruit and Shoot Borer- Leucinodes orbonalis Guenee is a major insect pest on brinjal- Solanum melongena worldwide. An effective strategy used in developing pest controlling agents is the synergism between insect pheromones and host plant volatiles, which can increase the attraction of insect pest. The present study was aimed at investigating the chemical constituents and attractant effects of the volatiles extracted from different parts of the host plant brinjal on the behavior of adult L. orbonalis. Bioassay using Y-shaped olfactometer revealed that the one-day old virgin female, gravid female and male insects respond positively to the host plant volatiles extracted from fruits, leaves and shoots but not to that of flowers. It was shown that the gravid females were significantly attracted to all three volatiles (p < 0.05). Bioassay using X-shaped olfactometer identified that all three types of insects highly preferred the volatiles from fruits (p < 0.05). Gas chromatography-mass spectrometry analysis of volatiles indicated that brinjal plant produces volatile secondary metabolites, which include 2,2′-(Ethane-1,2-diylbis(oxy))bis(ethane-2,1-diyl) dibenzoate (12.11%), 3,7-dimethylocta-1,6-dien-3-ol (22.38%), Benzyl alcohol (22.9%) and Benzyl alcohol (27.06%) as major constituents from fruits, shoots, leaves and flowers respectively. Responses of insects to the volatiles from host plant in the absence of visual cues direct us to focus on the importance of host plant volatiles to locate the plant. Results of this study emphasize the major role that host plant volatiles play in the attraction of insect pests towards the plant.     

Seasonal abundance and spatial distribution of larval and adult thrips (Thysanoptera) on weed host plants in mango orchards in Penang,Malaysia

The spatial distribution of larval and adult thrips (Thysanoptera) was studied on mango panicles, Mangifera indica L., on Penang Island, Malaysia, during two consecutive mango flowering seasons from December 2008 to March 2009 and from August to September 2009. Larval and adult thrips were sampled from mango panicles using the carbon dioxide (CO₂) collection technique weekly in treated and untreated orchards. Seasonal abundance and dispersion pattern of thrips were investigated on weed host plants in the treated orchard between June 2008 and March 2009. Spatiotemporal dynamics of larvae and adults was analyzed using Taylor’s Power Law (TPL), Lloyd’s Index (LI), and Green’s Index (GI). Thrips hawaiiensis (Morgan) was the dominant thrips species recovered from mango panicles and weeds in the treated orchard, whereas Scirtothrips dorsalis (Hood) was the most abundant species captured in the untreated orchard. Thrips adults and larvae analyzed via dispersion indices were found to be aggregated in mango panicles in both orchards. The value of the aggregation index (b) of TPL for the total number of adult thrips was significantly higher in the treated orchard than in the untreated orchard, whereas slopes of TPL for the total number of larval thrips did not differ significantly between treated and untreated orchards. All adult thrips species were distributed regularly on the weed plants; however, their larvae were aggregated. It is concluded that pesticide treatment caused adult thrips to become more aggregated on mango panicles, hiding in flower parts that were less exposed to the chemicals.     

Reproductive interference between two serious pests,oriental fruit flies <Emphasis Type="Italic">Bactrocera carambolae</Emphasis> and <Emphasis Type="Italic">B. dorsalis</Emphasis> (Diptera: Tephritidae), with very wide but partially overlapping host ranges

Bactrocera carambolae and B. dorsalis (Diptera: Tephritidae) are extremely destructive pests of fruits and vegetables in the Asia-Pacific region. Earlier reports have described that B. carambolae and B. dorsalis, respectively, use mainly star fruit and mango, suggesting a certain level of host partitioning which can be ascribed neither to differences in larval food qualities nor host-specific parasitoid mortality. This study specifically examined reproductive interference (antagonistic sexual interaction) between B. carambolae and B. dorsalis as a potential factor strongly affecting their host partitioning. We observed mating behaviors, especially interspecific courtships and mating, by cohabiting the conspecific and heterospecific pairs together. Consequently, we quantified their effects on the reproductive success of females. Males of both species frequently courted their own females, but they also courted females of other species. Courtship refusal by females was not selective in males of either species. This incomplete discrimination of both sexes led to frequent occurrences of interspecific sexual interactions in both species, but only B. carambolae females showed reduced reproductive success. These results suggest that B. dorsalis, superior in reproductive interference, can occupy high-quality mango, whereas B. carambolae, inferior in reproductive interference, must use low-quality star fruit.     

Molecular Characteristics,mRNA Expression,and Alternative Splicing of a Ryanodine Receptor Gene in the Oriental Fruit Fly,Bactrocera dorsalis (Hendel)

Ryanodine receptors (RyRs) are a distinct class of ligand-gated channels controlling the release of calcium from intracellular stores. The emergence of diamide insecticides, which selectively target insect RyRs, has promoted the study of insect RyRs. In the present study, the full-length RyR cDNA (BdRyR) was cloned and characterized from the oriental fruit fly, Bactrocera dorsalis (Hendel), a serious pest of fruits and vegetables throughout East Asia and the Pacific Rim. The cDNA of BdRyR contains a 15,420-bp open reading frame encoding 5,140 amino acids with a predicted molecular weight of 582.4 kDa and an isoelectric point of 5.38. BdRyR shows a high level of amino acid sequence identity (78 to 97%) to other insect RyR isoforms. All common structural features of the RyRs are present in the BdRyR, including a well-conserved C-terminal domain containing consensus calcium-binding EF-hands and six transmembrane domains, and a large N-terminal domain. Quantitative real-time PCR analyses revealed that BdRyR was expressed at the lowest and highest levels in egg and adult, respectively, and that the BdRyR expression levels in the third instar larva, pupa and adult were 166.99-, 157.56- and 808.56-fold higher, respectively, than that in the egg. Among different adult body parts, the highest expression level was observed in the thorax compared with the head and abdomen. In addition, four alternative splice sites were identified in the BdRyR gene, with the first, ASI, being located in the central part of the predicted second spore lysis A/RyR domain. Diagnostic PCR analyses revealed that alternative splice variants were generated not only in a tissue-specific manner but also in a developmentally regulated manner. These results lay the foundation for further understanding the structural and functional properties of BdRyR, and the molecular mechanisms for target site resistance in B. dorsalis.     

Additional measurements of distance-dependent capture probabilities for released males of Bactrocera cucurbitae and B. dorsalis (Diptera: Tephritidae) in Honolulu

This paper describes a mark-release-recapture study involving males of two economically important fruit flies (Diptera: Tephritidae), Bactrocera cucurbitae (Coquillett) and B. dorsalis (Hendel), conducted in Honolulu, Hawaii. In each of three residential neighborhoods in Honolulu, we placed two traps, one baited with cue lure and the other with methyl eugenol (male attractants for B. cucurbitae and B. dorsalis, respectively), in a single tree. For both species, dyed, mature males from recently established laboratory colonies were released 25, 50, 100, and 200 m from the traps along the four compass directions, and for B. dorsalis releases were also made 300 m from the traps. For both B. cucurbitae and B. dorsalis, the proportion of males captured varied significantly among study areas and among release distances. Averaged over the 3 sites, the percentage of released males captured varied from 14% for releases at 25 m to 0.5% for releases at 200 m for B. cucurbitae and from 21% for releases at 25 m to 3% for releases at 300 m for B. dorsalis. In general, the capture rates of B. cucurbitae males were similar between California and Hawaii, whereas the captures rates of B. dorsalis males were significantly higher in California than Hawaii for all releases distances tested. Distance-dependent capture rates were used to estimate detection sensitivities for the two species under a trapping regime utilizing 5 cue lure- and 5 methyl eugenol-baited traps per 2.59 km² (as currently employed in California) and then compare these sensitivities between California and Hawaii.     

Feeding of pea leafminer larvae simultaneously activates jasmonic and salicylic acid pathways in plants to release a terpenoid for indirect defense

The pea leafminer, Liriomyza huidobrensis, is an important pest species affecting ornamental crops worldwide. Plant damage consists of oviposition and feeding punctures created by female adult flies as well as larva-bored mines in leaf mesophyll tissues. How plants indirectly defend themselves from these two types of leafminer damage has not been sufficiently investigated. In this study, we compared the indirect defense responses of bean plants infested by either female adults or larvae. Puncturing of leaves by adults released green leaf volatiles and terpenoids, while larval feeding caused plants to additionally emit methyl salicylate and (E,E)-4,8,12-trimethyl-1,3,7,11-tridecatetraene (TMTT). Puncturing of plants by female adults induced increases in jasmonic acid (JA) and JA-related gene expressions but reduced the expressions of salicylic acid (SA)-related genes. In contrast, JA and SA and their-related gene expression levels were increased significantly by larval feeding. The exogenous application of JA+SA significantly triggered TMTT emission, thereby significantly inducing the orientation behavior of parasitoids. Our study has confirmed that larval feeding can trigger TMTT emission through the activation of both JA and SA pathways to attract parasitoids; however, TMTT alone is less attractive than the complete blend of volatiles released by infested plants.     

Host status of ‘Scifresh’ apples to the invasive fruit fly species Bactrocera dorsalis,Zeugodacus cucurbitae,and Ceratitis capitata (Diptera: Tephritidae)

We conducted no-choice cage and field infestation studies to determine if the fruit of apples (Malus x domestica L., ‘Scifresh’) are hosts for three invasive tephritid fruit fly species that may enter New Zealand or other apple growing areas. In screen cage tests, punctured and unpunctured (intact) fruit of ‘Scifresh’ apples were exposed to gravid females of Bactrocera dorsalis (Hendel) (Oriental fruit fly), Zeugodacus (Bactrocera) cucurbitae (Coquillet) (melon fly), or Ceratitis capitata (Wiedemann) (Mediterranean fruit fly), outdoors for 24 h and then held on sand in the laboratory for four weeks for pupal development and adult emergence. Unpunctured fruit produced an average of 269.4, 4.3 and 70.1 puparia per kg of fruit for B. dorsalis, Z. cucurbitae and C. capitata, respectively. Punctured fruit produced an average of 619.4, 0.8 and 129.5 puparia per kg of fruit for B. dorsalis, Z. cucurbitae and C. capitata, respectively. By comparison, unpunctured and punctured papaya fruit (Carica papaya, ‘Rainbow’, a preferred host) produced 206–675 and 464–735 puparia per kg of fruit, respectively, across all species. In general, the average weight of individual fruit fly puparia from apple was significantly less (41–71%) than that of puparia reared from papaya, and development times were slower on apple than on papaya. Overall, ‘Scifresh’ apples were a moderately good host for B. dorsalis and C. capitata, and a very poor host for Z. cucurbitae in cage tests. Field exposure of ‘Scifresh’ apples suspended from papaya trees resulted in no infestation by B. dorsalis or Z. cucurbitae under natural conditions. This information will help to inform decisions about quarantine restrictions and potential crop loss in the event of incursions of these fruit flies into apple-producing countries.     

Cytoplasmic glutamine synthetase gene expression regulates larval development in Bactrocera dorsalis (Hendel)

In insects, glutamine synthetase (GS), a key enzyme in the synthesis of glutamine, has been reported to be associated with embryonic development, heat shock response, and fecundity regulation. However, little is known about the influence of GS on postembryonic development. In this study, we demonstrate that blocking the activity of GS in the oriental fruit fly (Bactrocera dorsalis) with use of a GS‐specific inhibitor (L‐methionine S‐sulfoximine), led to a significant delay in larval development, pupal weight loss, and inhibition of pupation. We further identify cloned and characterized two GS genes (BdGS‐c and BdGS‐m) from B. dorsalis. The two GS genes identified in B. dorsalis were predicted to be located in the cytosol (BdGS‐c) and mitochondria (BdGS‐m), and homology analysis indicated that both genes were similar to homologs from other Dipterans, such as Drosophila melanogaster and Aedes aegypti. BdGS‐c was highly expressed in the larval stages, suggesting that cytosolic GS plays a predominant role in larval development. Furthermore, RNA interference experiments against BdGS‐c, to specifically decrease the expression of cytosolic GS, resulted in delay in larval development as well as pupal weight loss. This study presents the prominent role played by BdGS‐c in regulating larval development and suggests that the observed effect could have been modulated through ecdysteroid synthesis, agreeing with the reduced expression of the halloween gene spook. Also, the direct effects of BdGS‐c silencing on B. dorsalis, such as larval lethality, delayed pupation, and late emergence, can be further exploited as novel insecticide target in the context of pest management.