Adipokinetic hormone receptor gene identification and its role in triacylglycerol mobilization and sexual behavior in the oriental fruit fly (Bactrocera dorsalis)

Energy homeostasis requires continuous compensation for fluctuations in energy expenditure and availability of food resources. In insects, energy mobilization is under control of the adipokinetic hormone (AKH) where it is regulating the nutritional status by supporting the mobilization of lipids. In this study, we characterized the gene coding for the AKH receptor (AKHR) and investigated its function in the oriental fruit fly (Bactrocera dorsalis) that is economically one of the most important pest insects of tropical and subtropical fruit. Bacdo-AKHR is a typical G protein-coupled receptor (GPCR) and phylogenetic analysis confirmed that Bacdo-AKHR is closely related to insect AKHRs from other species. When expressed in Chinese hamster ovary (CHO) cells, Bacdo-AKHR exhibited a high sensitivity and selectivity for AKH peptide (EC₅₀ = 19.3 nM). Using qPCR, the developmental stage and tissue-specific expression profiles demonstrated that Bacdo-AKHR was highly expressed in both the larval and adult stages, and also specifically in the fat body and midgut of the adult with no difference in sex. To investigate the role of AKHR in B. dorsalis, RNAi assays were performed with dsRNA against Bacdo-AKHR in adult flies of both sexes and under starvation and feeding condition. As major results, the knockdown of this gene resulted in triacylglycerol (TAG) accumulation. With RNAi-males, we observed a severe decrease in their sexual courtship activity when starved, but there was a partial rescue in copulation when refed. Also in RNAi-males, the tethered-flight duration declined compared with the control group when starved, which is confirming the dependency on energy metabolism. In RNAi-females, the sexual behavior was not affected, but their fecundity was decreased. Our findings indicate an interesting role of AKHR in the sexual behavior of males specifically. The effects are associated with TAG accumulation, and we also reported that the conserved role of AKH-mediated system in B. dorsalis is nutritional state-dependent. Hence, we provided further understanding on the multiple functions of AKH/AKHR in B. dorsalis.     

Low doses of a neonicotinoid insecticide modify pheromone response thresholds of central but not peripheral olfactory neurons in a pest insect

Insect pest management relies mainly on neurotoxic insecticides, including neonicotinoids, leaving residues in the environment. There is now evidence that low doses of insecticides can have positive effects on pest insects by enhancing various life traits. Because pest insects often rely on sex pheromones for reproduction, and olfactory synaptic transmission is cholinergic, neonicotinoid residues could modify chemical communication. We recently showed that treatments with different sublethal doses of clothianidin could either enhance or decrease behavioural sex pheromone responses in the male moth, Agrotis ipsilon. We investigated now effects of the behaviourally active clothianidin doses on the sensitivity of the peripheral and central olfactory system. We show with extracellular recordings that both tested clothianidin doses do not influence pheromone responses in olfactory receptor neurons. Similarly, in vivo optical imaging does not reveal any changes in glomerular response intensities to the sex pheromone after clothianidin treatments. The sensitivity of intracellularly recorded antennal lobe output neurons, however, is upregulated by a lethal dose 20 times and downregulated by a dose 10 times lower than the lethal dose 0. This correlates with the changes of behavioural responses after clothianidin treatment and suggests the antennal lobe as neural substrate involved in clothianidin-induced behavioural changes.     

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.     

Reduced insecticide susceptibility among populations of Scirtothrips dorsalis Hood (Thysanoptera: Thripidae) in strawberry production

Chilli thrips, Scirtothrips dorsalis Hood (Thysanoptera: Thripidae) is a major, economically important, and recent invasive pest of strawberries and other horticultural crops in United States. Several conventional insecticides are used for S. dorsalis management, and resistance development threatens loss of few available tools. Hence, our objectives were to: (1) determine the susceptibility of S. dorsalis to commonly used conventional insecticides: spinetoram, acetamiprid, cyantraniliprole and bifenthrin, and (2) establish LC₅₀ and LC₉₀ dosages for spinetoram against S. dorsalis. Sampling of S. dorsalis populations was conducted twice in seven strawberry fields in Florida during the strawberry field season between 2019 and 2020. Leaf-disc bioassays were performed with field collected populations along with a susceptible 2-year-old laboratory culture of S. dorsalis. Overall, at highest recommended rate the percent mortality of late season S. dorsalis populations from five out of seven collection sites was lower (~41%) than average mortality observed with early season populations (~72%). Populations from at least four out of seven sampling sites exhibited significantly lower mortality than the laboratory susceptible culture in late season. The LC₅₀ and LC₉₀ values for spinetoram for the susceptible laboratory population were 0.026 and 8.64 ppm, respectively. On the other hand, LC₅₀ values of field collected populations to spinetoram varied with resistance ratios ranging from 6 to 269 fold as compared against the laboratory strain. Our results suggest that susceptibility of S. dorsalis to commonly used insecticides in strawberries varies significantly between early and late season populations within the same crop season. The efficacy of bifenthrin against S. dorsalis was particularly low (~ reduced to half in late season), especially among field collected populations. Our results indicate an urgent need to incorporate other pest management strategies, as well as effective rotation programs to reduce selection for resistance among populations of S. dorsalis in strawberry production.     

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.     

The photokinesis of oriental fruit flies,Bactrocera dorsalis,to LED lights of various wavelengths

The oriental fruit fly, Bactrocera dorsalis Hendel (Diptera: Tephritidae), is an invasive pest of orchards around the world, particularly in Asian countries such as China. Light traps offer a potential means for pest monitoring and management. This study aimed to evaluate the sensitivity of the fly to light and investigate the impact of monochromatic light in the sensitivity spectrum on B. dorsalis. Six light wavelengths in LEDs – green (522 nm), yellow (596 nm), blue (450 nm), red (633 nm), purple (440 nm), and white (compound light) – were adapted to test responses of 5‐, 10‐, and 20‐day‐old B. dorsalis adults kept in laboratory conditions. We also tested the effects of green and red lights on pupal development and adults’ life activities. The results indicated a phototaxis preference rank in B. dorsalis adults to monochromatic LEDs with, in decreasing order, green, yellow, purple, blue, and red. Moreover, positive phototaxis significantly increased with age. Male adults are more sensitive than female adults to test lights, mainly at the age of 10 and 20 days. Emergence rates of pupae exposed to 12 and 24 h green light daily were 42 and 67%, respectively, whereas controls held in red light emerged at 33 and 37%, respectively. Furthermore, body weight, female fecundity, and mortality of B. dorsalis in night‐time exposure of green light (from 21:00 to 09:00 hours; during daytime flies were illuminated by white LED light) were significantly higher than in red‐light test groups and dark controls. In conclusion, B. dorsalis displayed preference toward green light, and fly age and gender seemed to significantly impact the phototactic behavior. Green LED light exposure during nighttime remarkably improved the emergence rates of B. dorsalis, and it enhanced growth, development, and ovipositing peak period, but decreased adult lifespan. This research lays a foundation for the development of new trap models, e.g., with green sticky cards or green light, for monitoring and control of B. dorsalis in the field.     

Antennal lobe organization in the slender pigeon louse,Columbicola columbae (Phthiraptera: Ischnocera)

This study reports on the structure of the antennal lobe of the pigeon louse, Columbicola columbae. Anterograde staining of antennal receptor neurons revealed an antennal lobe with a few diffuse compartments, an organization distinct from the typical spheroidal glomerular structure found in the olfactory bulb of vertebrates and the antennal lobe of many other insects. This anatomical arrangement of neuronal input is somewhat reminiscent of the aglomerular antennal lobe previously reported in psyllids and aphids. As in psyllids, reports on the odor-mediated behavior of C. columbae suggest that the olfactory sense is important in these animals and indicates that a glomerular organization of the antennal lobe may not be necessary to subtend odor-mediated behaviors in all insects. The diffuse or aglomerular antennal lobe organization found in these two Paraneopteran insect orders might represent an independently evolved reduction due to similar ecological constraints.     

Juvenile hormone regulation on the flight capability of Bactrocera dorsalis (Diptera: Tephritidae)

The oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), is considered a major economic threat in many regions worldwide. In order to better understand the flight capacity of B. dorsalis and its physiological basis, the functions and regulatory roles of juvenile hormone (JH) in the flight muscle of B. dorsalis were studied under a controlled environment. JH titer of B. dorsalis varied with age and sex. Females have higher JH titers and better flight capabilities than males, given that the increase in JH also corresponded to the ovarian development and maturation in females. The flight duration and distance of both males and females increased with the gradual increase of JH titer after adult emergences. Both JH titer and flight capability peaked in 15-d-old adult and declined subsequently with age. Flight activity stimulated the production of JH as adults flown for 24 h on the flight mills have the highest JH titers compare to adults flown on shorter flight durations. Adults treated with 0.5 µg and 5 µg of JH III were able to fly long durations and long distances, nevertheless when JH titer was too low or too high, it would restrict the flight ability of the fly. The mutual reinforcement of JH and flight activity provides fundamental understanding on the physiological aspects of the flight capability and dispersal, which facilitates strategies for the long-term control of this destructive pest.     

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.     

Population genetic structure of Bactrocera dorsalis based on cox1 sequences from Bangladesh and neighboring countries

Oriental fruit fly, Bactrocera dorsalis (Hendel), is a destructive and highly polyphagous invasive fruit fly species of numerous fruit crops in global agriculture. Population genetic structure of this species from five different locations of Bangladesh was examined with other samples (collected from GenBank) from 15 sites of neighboring Asian countries. A fragment of 770 bp mitochondrial DNA cox1 was used to investigate the genetic diversity and the relationship between genetic patterns and geographical distribution of B. dorsalis. A total of 232 variable sites (33.23% of the 698 bp aligned consensus sequences) and 419 unique haplotypes were identified from 710 individuals. Indices of genetic diversity suggested that without exclusion from geographical areas, B. dorsalis retained a relatively high degree of genetic diversity. A demographic assessment [Tajimas’ D test, Fu’s Fs test and sum of square deviation (SSD values)] revealed that both current and historical variables performed a significant role in deciding the weak genetic structure with some exceptions. In Bangladesh, high levels of genetic diversity with a weak genetic structure indicated that the severity of this pest might increase in the future. Proper management techniques should be taken to overcome the future severity of this kind of destructive insect.     

应用寄生蜂和不育雄虫防控田间橘小实蝇

食品安全已成为当今社会广受关注的问题。尽管国内的一些学者就阿里山潜蝇茧蜂[Fopius arisanus(Sonan)]和橘小实蝇SIT技术分别开展了不同程度的研究,然而关于应用该蜂和不育雄虫对橘小实蝇[Bactrocera dorsalis(Hendel)]进行田间联合防治的研究还未见相关报道。为此,为探讨天敌昆虫和雄性不育技术对橘小实蝇的田间综合治理(IPM)技术,开展了阿里山潜蝇茧蜂和橘小实蝇不育雄虫的田间释放试验:首先应用性引诱剂降低田间野生雄虫数量;而后,释放橘小实蝇不育雄虫,进一步降低田间橘小实蝇种群数量;最后,释放阿里山潜蝇茧蜂对田间残余的橘小实蝇卵进行追踪寄生,以达到持续控制的目的。通过监测整个防控期田间橘小实蝇的种群数量以及果实受害率和防治效果作为评价指标评价该防治方法的综合防治效果。结果表明:应用阿里山潜蝇茧蜂、橘小实蝇不育雄虫并配合引诱剂的综合措施对橘小实蝇的种群数量有明显控制作用,田间总体防治效果可达90%左右。此外,提出完善橘小实蝇田间防控措施的建议,并提出该研究的不足以及有待进一步开展研究的方向。     

Salicylic Acid Induces Changes in Mango Fruit that Affect Oviposition Behavior and Development of the Oriental Fruit Fly,Bactrocera dorsalis

The Oriental fruit fly, Bactrocera dorsalis (Hendel) is an important quarantine pest around the globe. Although measures for its control are implemented worldwide through IPM and male annihilation, there is little effect on their population. Hence, there is a need for new strategies to control this minacious pest. A strategy that has received negligible attention is the induction of ‘natural plant defenses’ by phytohormones. In this study, we investigated the effect of salicylic acid (SA) treatment of mango fruit (cv. Totapuri) on oviposition and larval development of B. dorsalis. In oviposition choice assays, gravid females laid significantly less eggs in SA treated compared to untreated fruit. Headspace volatiles collected from SA treated fruit were less attractive to gravid females compared to volatiles from untreated fruit. GC-MS analysis of the headspace volatiles from SA treated and untreated fruit showed noticeable changes in their chemical compositions. Cis-ocimene and 3-carene (attractants to B. dorsalis) were reduced in the headspace volatiles of treated fruit. Further, reduced pupae formation and adult emergence was observed in treated fruit compared to control. Increased phenol and flavonoid content was recorded in treated fruit. We also observed differential expression of anti-oxidative enzymes namely catalase (CAT), polyphenoloxidase (PPO) and peroxidase (POD). In summary, the results indicate that SA treatment reduced oviposition, larval development and adult emergence of B. dorsalis and suggest a role of SA in enhancing mango tolerance to B. dorsalis.     

Morphological and ultrastructural characterization of olfactory sensilla in Drosophila suzukii: Scanning and transmission electron microscopy

Drosophila suzukii is a serious horticultural and quarantine pest, damaging various berry crops. Although the active use of olfactory communication in D. suzukii is well-known, their olfactory sensory system has not been comprehensively reported. Therefore, the present study was carried out to understand the morphology, distribution and ultrastructure of olfactory sensilla present in the antennae and maxillary palps of D. suzukii, through scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The olfactory sensilla on the antennae of D. suzukii in both sexes could be classified into three major morphological types, basiconic, trichoid and coeloconic sensilla, according to their shapes. The antennal basiconic sensilla were further divided into three subtypes and the antennal trichoid sensilla into two subtypes, respectively, according to the size of individual sensillum. In contrast to the antennal olfactory sensilla showing diverse morphology, basiconic sensilla was the only type of olfactory sensilla in the maxillary palps of D. suzukii. The basiconic sensilla in the maxillary palps could be further classified into three subtypes, based on their size. Our SEM and TEM observations indicated that multiple nanoscale pores are present on the surface of all types of olfactory sensilla in the antennae and maxillary palps, except coeloconic sensilla. The difference in the morphological types and the distribution of olfactory sensilla suggests that their olfactory functions are different between antennae and maxillary palps in D. suzukii. The results of this study provide useful information for further studies to determine the function of olfactory sensilla in D. suzukii and to understand their chemical communication system.     

Population structure and colonization of Bactrocera dorsalis (Diptera: Tephritidae) in China,inferred from mtDNA COI sequences

The oriental fruit fly, Bactrocera dorsalis, is a serious pest of fruits and vegetables in South‐east Asia, and, because of quarantine restrictions, impedes international trade and economic development in the region. Revealing genetic variation in oriental fruit fly populations will provide a better understanding of the colonization process and facilitate the quarantine and management of this species. The genetic structure in 15 populations of oriental fruit fly from southern China, Laos and Myanmar in South‐east Asia was examined with a 640‐bp sequence of the mitochondrial cytochrome oxidase subunit I (COI) gene. The highest levels of genetic diversity were found in Laos and Myanmar. Low to medium levels of genetic differentiation (F_ST ≤ 0.134) were observed among populations. Pooled populations from mainland China differed from those in Laos and Myanmar (F_ST = 0.024). Genetic structure across the region did not follow the isolation‐by‐distance model. The high genetic diversity observed in Laos and Myanmar supports the South‐east Asian origin of B. dorsalis. High genetic diversity and significant differentiation between some populations within mainland China indicate B. dorsalis populations have been established in the region for an extended period of time. High levels of genetic diversity observed among the five populations from Hainan Island and similarity between the Island and Chinese mainland populations indicate that B. dorsalis was introduced to Hainan from the mainland and has been on the island for many years. High genetic diversity in the recently established population in Shanghai (Pudong) suggests multiple introductions or a larger number of founders.     

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.