The role of cAMP-dependent protein kinase A in the formation of long-term memory in Bactrocera dorsalis

The cAMP-dependent protein kinase A(PKA)signaling pathway has long been considered critical for long-term memory(LTM)formation.Previous studies have mostly focused on the role of PKA signaling in LTM induction by multiple spaced conditioning with less attention to LTM induction by a single conditioning.Here,we conducted behavioral-pharmacology,enzyme immunoassay and RNA interference experiments to study the role of the PKA signaling pathway in LTM formation in the agricultural pest Bactrocera dorsalis,which has a strong memory capacity allowing it to form a two-day memory even from a single conditioning trial.We found that either blocking or activating PKA prior to conditioning pretreatment affected multiple spaced LTM,and conversely,they did not affect LTM formed by single conditioning.This was further confirmed by enzyme-linked immunosorbent assay(ELISA)and silencing of the protein kinase regulatory subunit 2 and catalytic subunit 1.Taken together,these results suggest that activating PKA during memory acquisition helps to induce the LTM formed by multiple spaced conditioning but not by a single conditioning.Our findings challenge the conserved role of PKA signaling in LTM,which provides a basis for the greater diversity of molecular mechanisms underlying LTM formation across species,as well as possible functional and evolutionary implications.

Characterization of Domeless receptors and the role of Bd Domeless3 in anti-symbiont-like virus defense in Bactrocera dorsalis

The Janus kinase/signal transducers and activators of transcription(JAK/STAT)signaling pathway play a pivotal role in innate immunity.Among invertebrates,Domeless receptors serve as the key upstream regulators of this pathway.In our study on Bactrocera dorsalis,we identified three cytokine receptors:BdDomeless1,BdDomeless2,and BdDomeless3.Each receptor encompasses five fibronectin-type-III-like(FN III)extracellular domains and a transmembrane domain.Furthermore,these receptors exhibit the increased responsiveness to diverse pathogenic challenges.Notably,only BdDomeless3 is upregulated during symbiont-like viral infections.Moreover,silencing BdDomeless3 enhanced the infectivity of Bactrocera dorsalis cripavirus(BdCV)and B.dorsalis picorna-like virus(BdPLV),underscoring BdDomeless3’s crucial role in antiviral defense of B.dorsalis.Following the suppression of Domeless3 expression,six antimicrobial peptide genes displayed decreased expression,potentially correlating with the rise in viral infectivity.To our knowledge,this is the first study identifying cytokine receptors associated with the JAK/STAT pathway in tephritid flies,shedding light on the immune mechanisms of B.dorsalis.

The Essential Oil of Ocimum americanum from Senegal and Gambia as a Source of Methyleugenol for the Control of Bactrocera dorsalis, Fruit Fly

The fruit fly, Bactrocera dorsalis (Diptera: Tephritidae) is one of the most important pests in all mango-producing areas, particularly in West Africa. In Senegal, O. americanum leaves have been used for several years to control this fly. However, to our knowledge, no chemical studies have been carried out. Thus, the aim of this study is to determine the chemical composition of the essential oil of O. americanum leaves collected in Senegal and Gambia. The essential oil obtained by hydrodistillation of these leaves is analyzed by GC/FID and GC/MS. Yields of essential oils from O. americanum leaves are 3.84% and 2.13%, respectively. Analysis of these essential oils by GC/FID and GC/MS allowed the identification of 23 compounds representing almost 100% of the total compositions. These essential oils are mainly dominated by methyleugenol (72.0% and 75.8%, respectively). Other components in significant percent are trans-β-caryophyllene (13.9% and 13.0%, respectively), germacrene D (4.1% and 3.7%, respectively), β-elemene (3.3% and 0.9%, respectively). Due to the high methyleugenol content, this study explains the attractive potential of O. americanum towards B. dorsalis. In perspective, we plan to evaluate the attractive effect of the essential oil and leaf powder of O. americanum against B. dorsalis, a real pest of mango orchards in Senegal.

Light intensity regulates the sexual behaviors of oriental fruit fly Bactrocera dorsalis under laboratory conditions

The oriental fruit fly,Bactrocera dorsalis(Hendel),is a devastating pest of citrus fruits.After successful mating,adult females insert their eggs into the ripened fruit,resulting in moldy and rotten fruit and causing great economic losses for the citrus industry.In the field,flies initiate copulatory behaviors as twilight approaches,and decreasing light intensity in this period is the normal stimulus for copulation.In this study,ten light intensities ranging from 0–30000 lux were set to identify the typical intensity that strongly regulates the copulation behavior of B.dorsalis.Three light intensities found to regulate the copulation behavior were then selected to verify their effects on adult male wing fanning and female chemotaxis towards 2,3,5-trimethylpyrazine(TMP).At last,strong light and complete darkness were artificially combined in the lab to verify whether they could prevent copulation to inform behavioral manipulation of oriental flies in the future.The results indicated that adult flies generally initiated copulatory behaviors at low light intensity(<1000 lux).Stronger light significantly prevented copulation in proportion to intensity,with nearly no copulation events initiated when light intensity was above 20000 lux.Both male wing fanning and female chemotaxis towards TMP were attenuated as light intensity became stronger.However,at 10000 lux,males still fanned their wings to a certain extent while TMP completely lost its attractiveness to females.In the darkness,adults did not initiate any sexual behaviors,e.g.,copulation,wing fanning,or chemotaxis to TMP.One hour of strong light(10000 lux)combined with continuous darkness completely prevented mating.These results show that light condition is an essential factor for copulatory behaviors in the oriental fruit fly.Researchers could thus manipulate light conditions artificially or disrupt the molecular target in flies’light transduction pathway to develop environmentally-friendly techniques to control this pest.

Research Progress on the Innate Immunity of Bactrocera dorsalis (Hendel) and the Regulation Mechanism of Parasitic Wasp Venom Protein

[Objectives]The use of natural enemies of living insects and their derivatives can effectively avoid the problems of pesticide residues,pest resistance,biodiversity decline,control effect weakening and so on.[Methods]Parasites inject various parasitic factors into hosts to inhibit the development of hosts,adjust the immunity of hosts,interfere with the growth and development of hosts,and reduce the nutrition metabolism of hosts,so as to ensure the growth and development of the offspring.Host pests can escape or conquer the parasitism of parasitic wasps through immune defense system in order to reproduce their own offspring.[Results]Under intense and strong selection pressure,in order to effectively ensure the success rate of parasitism,the adaptive diversity of parasitism strategies of parasitic wasps is finally caused.In the process of evolution and under the pressure of directional selection,the innate immunity and acquired immunity gradually evolve.[Conclusions]In-depth research on parasitic factors of parasitic wasps and their interaction with crop pests immunity and development can not only improve theoretical understanding of insect immunity and development biology,pest biological control and other disciplines,but also be expected to enable the application of some components of parasitic factors to agriculture,medicine and pharmacy.Bactrocera dorsalis is a destructive fruit and vegetable pest.This paper summarized the venom protein of B.dorsalis parasitoids and the immune interaction with hosts,in order to provide theoretical basis for biological control of plant pests by using parasitic natural enemies.

玉溪市新平县桔小实蝇成虫种群消长特征及其与气象因子的关系

为了揭示玉溪市新平县桔小实蝇[Bactrocera dorsalis(Hendel)]成虫种群消长特征及其与气象因子的关系,于2020—2022年采用甲基丁香酚为诱芯的诱捕器,监测新平县桔小实蝇雄成虫的消长动态,并根据当地气象资料分析气象因子与桔小实蝇雄成虫数量的关系。结果表明,桔小实蝇在新平县全年皆发生,冬季成虫数量较少,夏季为桔小实蝇成虫暴发期,5月以后种群数量开始增长,种群数量暴发期为6—10月。气象因子显著影响了桔小实蝇雄虫的数量,其中主要受月平均气温和月平均最低气温的影响。由此可知,影响新平县桔小实蝇种群数量变动的主要因素是气温、降雨、寄主种类和海拔等,这些因素对桔小实蝇的作用方式、强度和时间各不相同,在成虫种群消长的不同阶段发挥不同作用,但通过特定时空下的相互组合,综合决定着新平县桔小实蝇种群消长的基本特征。

Relationship between Population Fluctuation of Oriental Fruit Fly Bactrocera dorsalis Hendel and Abiotic Factors in Yezin, Myanmar

Population fluctuation of oriental fruit fly, Bactrocera dorsalis （Hendel） （Diptera： Tephritidae）, was monitored by using methyl eugenol traps during May, 2011 to December, 2011 and May, 2016 to December, 2016 in mango orchard farms of Department of Agricultural Research in Yezin, Myanmar. In 2011, the highest mean number of male oriental fruit flies/trap/day （241.42 ± 11.79） was recorded in June and the lowest （2.21 ± 0.40） in December. The highest mean number of male oriental frui t flies/trap/day （388.95 ± 8.56） was recorded in the month of June and the lowest （2.33 ± 0.20） in December, 2016. Population data were correlated with meteorological data including temperature, duration of sunshine, and rainfall. Population fluctuation of male fruit flies was positively correlated with temperature and rainfall, and negatively correlated with the duration of sunshine.

Influence of Host Shift on Genetic Differentiation of the Oriental Fruit Fly,Bactrocera dorsalis

Invasion of the oriental fruit lfy, Bactrocera dorsalis, into new niches containing different food sources （a process referred to as host shift）, may cause population genetic differentiation and sympatric speciation. To attempt to infer that experimentally, test populations were established by transferring a subset of the original populations, which had been grown on banana for many generations, onto navel orange, and then subculturing the navel orange population and banana population for at least 20 generations. Four pairs of SSR primers with high polymorphism on laboratory strains were used to detect population genetic differentiation. All six tested populations （the 5th, 10th and 15th generations of B. dorsalis fed on banana and navel orange, respectively） were found to have low genetic diversity. Furthermore, the genetic diversity of the navel orange populations was found to decline after being crossed for several generations. Populations initially were deviated from Hardy-Weinberg equilibrium, however, equilibrium was achieved with increasing numbers of generations in both of the host populations. Limited gene lfows were found among the six populations. The Nei’s standard genetic distances between the two host populations of the same generation were initially low, but increased with generation number. Genetic distances between banana and navel orange populations of the same generation were lower than genetic distances between different generations grown on the same host plant. Analysis of molecular distance （AMOVA） results based on generation groups and host groups demonstrated that genetic variation among generations was greater than that between the two host populations. The results indicated that population genetic differentiation occurred after the host shift, albeit at low level. Biogeography and taxonomy of the B. dorsalis complex revealed that speciation of B. dorsalis might be tightly associated with host shift or host specialization of B. dorsalis following dispersal.

Trapping Effects of Different Treatments on Bactrocera dorsalis and Bactrocera cucurbitae

Five different trapping treatments, spraying attractant on bottle surface and spraying attractant inside the bottles with 0, 50,100 and 200 mL water, were set to trap Bactrocera dorsalis and B. cucurbitae in guava （Psidium guajava） park. The results showed that when the usage of attractant was 1 g, both Haonian and Wende had trapping effect on B. dorsalis and B. cucurbitae. The trapping effect of Haonian on B. dorsalis was better than that of Wende, while their trapping effects on B. cucurbitae was just the opposite. The trapping effects of different treatments had great difference. The trapping effect of Haonian on two species of fruit flies enhanced with the increasing volume of water, and reached the ma^mum value as the water volume was 200 mL. With the increasing volume of water, the trap- ping effect of Wende on two species of fruit flies first increased, and then decreased, which reached the maximum value as the water volume was 50 mL. Different treatments with attractants spraying inside bottles had better trapping effects on two species of fruit flies than that spraying on bottle surface.

Advances in Detection Methods for Invasive Pest Bactrocera dorsalis

Bactrocera dorsalis Hendel （Diptera： Tephritidae） is an invasive pest around the world. The paper summarizes biological and ecological characteristics of B, dorsalis, and reviews its detection methods from the aspects of morphological identification, acoustic detection and molecular detection, in order to provide a reference for further research and development of new detection methods. The hot issues in the study of B. dorsalis, such as ecological adaptation pattern, diffusion pathways and mechanisms, sustainable control measures, are also put forward in the paper.

Monitoring and Control Technology of Bactrocera dorsalis

After systematic monitoring and field survey in orchard for three years from 2009 to 2011, it was found that Bactrocera dorsalis occurred for more than three generations each year on the north shore of the middle and lower reaches of Yangtze River, with overlapping generations; the activity peak of adults was from mid August to late September; the occurrence in citrus orchard in hilly and mountainous area was severe; the average insect-fruit rate in October reached 0.20% - 15.88% ; B. dorsalis had sporadic occurrence in other orchards. The integrated control measures of plowing in winter to kill pupae, removing insect-fruits, boycott- ing on the ground, trapping by sex attraetants and spraying pesticides could basically control the damage of B. dorsalis.

点喷饵剂对芒果橘小实蝇的防治效果

为探讨芒果园中饵剂对橘小实蝇的防治效果及施药时期,在芒果成熟期监测了橘小实蝇[Bactrocera dorsalis(Hendel)]成虫发生动态,比较阿维菌素0.1%饵剂不同剂量对芒果的保护效果。结果表明,橘小实蝇发生高峰期与芒果成熟期相吻合,成熟前果园中橘小实蝇的密度较低,诱捕到成虫数低于60头/诱捕器,开始有黄熟芒果或落地果时,虫量快速增加,最高为1082.3头/诱捕器,采收结束后,虫口快速下降。使用阿维菌素0.1%饵剂2250、2700、3150 g/hm^(2)点喷,对芒果具有较好的保护效果,大部分处理与对照药剂2.5%高效氯氟氰菊酯水乳剂稀释1500倍液的防效差异不显著,对树上果的防控效果最高为82.1%,落地果为87.5%,采后果为79.5%。阿维菌素0.1%饵剂推荐使用方法为虫口暴发初期开始施药,使用剂量为3150 g/hm^(2),对植株点喷,每株果树喷药面积为1.0~1.5 m^(2),避免果实接触药液,间隔7 d施用,直至采收结束。

Identification and functional verification of Y-chromosome-specific gene typo-gyf in Bactrocera dorsalis

Genes on the Y chromosome play important roles in male sex determination and development.The identification of Y-chromosome-specific genes not only provides a theoretical basis for the study of male reproductive development,but also offers genetic control targets for agricultural pests.However,Y-chromosome genes are rarely characterized due to their high repeatability and high heterochromatinization,especially in the oriental fruit fly.In this study,1011 Y-chromosome-specific candidate sequences were screened from 2 to 4 h Bactrocera dorsalis embryo datasets with the chromosome quotient method,6 of which were identified as Y-chromosome-specific sequences by polymerase chain reaction,including typo-gyf,a 19126-bp DNA sequence containing a 575-amino acid open reading frame.Testicular deformation and a significant reduction in sperm number were observed after typo-gyf knockdown with RNA interference in embryos.After typo-gyf knockout with clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9 in the embryonic stage,the sex ratio of the emergent adults was unbalanced,with far more females than males.A genotype analysis of these females with the Y-chromosome gene MoY revealed no sex reversal.Typo-gyf knockout led to the death of XY individuals in the embryonic stage.We conclude that typo-gyf is an essential gene for male survival,and is also involved in testicular development and spermatogenesis.The identification of typo-gyf and its functional verification provide insight into the roles of Y-chromosome genes in male development.

性诱剂防治桔小实蝇的效果评价

采用生命表的干扰控制指数(IIPC)方法,就性诱剂对桔小实蝇(Bactrocera(Bactrocera)dorsalis(Hendel))种群的控制作用进行了评价.结果表明,性诱剂防治桔小实蝇的IIPC值为0 40.在低、中、高不同虫口密度的果园里,其保果率分别为73 60%、46 60%和23 00%.

百香果对桔小实蝇生长发育的影响

通过田间调查和室内试验探究桔小实蝇(Bactrocera dorsalis Hendel)对百香果的危害及不同成熟状态的百香果及百香果的不同部位对桔小实蝇生长发育的影响。田间调查结果显示,广东、广西百香果种植园内均能发现桔小实蝇成虫,且百香果果皮处可见为害痕迹,但果肉完整,果瓤内未发现虫体为害,51.7%的受害果被害部位发现实蝇卵。室内试验结果显示,桔小实蝇能在百香果上产卵,产卵痕数量平均为1.73个/每果,幼虫化蛹率为94.2%。桔小实蝇不能在成熟果的果皮上完成生活史,相比于果皮,百香果果肉显著提高了桔小实蝇的化蛹率和羽化率,但仍然低于香蕉对照组。成熟百香果的糖度为17.72%,pH为1.78,均不利于桔小实蝇的产卵和生长发育。综上所述,桔小实蝇能够为害百香果并能在果实上完成生长发育,但百香果不是桔小实蝇的适宜寄主。

Potential Geographic Distribution of Oriental Fruit Fly in Jiangxi Province

[Objective] The aim was to effectively monitor and manage oriental fruit fly.[Method] Maxent model and ArcGIS were used to predict potential geographic distributions of the oriental fruit fly based on associations between known occurrence records and a set of environmental variables.[Result] The suitable areas for Bactrocera dorsalis infestations were mainly restricted to central and southern Jiangxi Province,with Latitude ranged from 24 to 28°N.Northeast Jiangxi,Northwest Jiangxi and the regions bording upon Yangtse River were predicted as unsuitable for Bactrocera dorsalis.The fit for the model as measured by AUC was high,with value of 0.978 for the training data and 0.965 for the test data,indicating the high level of discriminatory power for the Maxent.A jackknife test in Maxent indicated that mean temperature of coldest quarter with highest gain value was the most important environmental variable that restricted the expansion to north Jiangxi Province.[Conclusion] Further research into the biology of the species and their ability to overcome barriers was necessary to explain niche differentiation and better understand invasion risk.

Formation characteristics of long-term memory in Bactrocera dorsalis

Studies on insects have contributed significantly to a better understanding of learning and memory,which is a necessary cognitive capability for all animals.Although the formation of memory has been studied in some model insects,more evidence is required to clarify the characteristics of memory formation,especially long-term memory(LTM),which is important for reliably storing information.Here,we explored this question by examining Bactrocera dorsalis,an agricultural pest with excellent learning abilities.Using the classical conditioning paradigm of the olfactory proboscis extension reflex(PER),we found that paired conditioning with multiple trials(>3)spaced with an intertrial interval(≥10 min)resulted in stable memory that lasted for at least 3 d.Furthermore,even a single conditioning trial was sufficient for the formation of a 2-d memory.With the injection of protein inhibitors,protein-synthesis-dependent memory was confirmed to start 4 h after training,and its dependence on translation and transcription differed.Moreover,the results revealed that the dependence of memory on protein translation exhibited a time-window effect(4-6 h).Our findings provide an integrated view of LTM in insects,suggesting common mechanisms in LTM formation that play a key role in the biological basis of memory.

Integrated analysis of miRNA and mRNA expression profiles in response to gut microbiota depletion in the abdomens of female Bactrocera dorsalis

Insect gut microbiota has been reported to participate in regulating host multiple biological processes including metabolism and reproduction.However,the corresponding molecular mechanisms remain largely unknown.Recent studies suggest that microRNAs(miRNAs)are involved in complex interactions between the gut microbiota and the host.Here,we used next-generation sequencing technology to characterize miRNA and mRNA expression profiles and construct the miRNA–gene regulatory network in response to gut microbiota depletion in the abdomens of female Bactrocera dorsalis.A total of 3016 differentially expressed genes(DEGs)and 18 differentially expressed miRNAs(DEMs)were identified.Based on the integrated analysis of miRNA and mRNA sequencing data,229 negatively correlated miRNA-gene pairs were identified from the miRNA-mRNA network.Gene ontology enrichment analysis indicated that DEMs could target several genes involved in the metabolic process,oxidation-reduction process,oogenesis,and insulin signaling pathway.Finally,real-time quantitative polymerase chain reaction further verified the accuracy of RNA sequencing results.In conclusion,our study provides the profiles of miRNA and mRNA expressions under antibiotics treatment and provides an insight into the roles of miRNAs and their target genes in the interaction between the gut microbiota and its host.

Population dynamics of the oriental fruit fly, Bactrocera dorsalis （Diptera： Tephritidae） in the Kunming area,southwestern China

Population dynamics of Bactrocera dorsalis Hendel （Diptera： Tephritidae） were studied through pheromone trapping over 4 years （1997, 1999, 2000, 2003） in the Kunming region, a high plateau area in southwestern China. B. dorsalis immigrates from southern Yunnan to Kunming each year, and occurs during early May through November. Annual trap captures recorded an increase in the B. dorsalis populations from May to July, when they peaked in abundance, and a decline until November. No flies were detected from November to April. The fruit flies had two generations. There was considerable overlapping due to the continuous arrival of immigrating flies during the summer months. Annual capture rates were significantly related to numbers of flies caught in July when peak captures were recorded; whereas the peak captures, in turn, positively depended on numbers of flies recorded in May, the first month of fly appearance in the current year. It suggested that the annual population abundance was mainly dependent on the size of the initial emigrating population. A daily average temperature of 18℃ was probably the threshold temperature required for the flies to undertake long-range dispersal, which partially explained the start of the fly in May each year on this high plateau. Under field conditions, the fruit flies can withstand 13℃ as a daily average temperature. No flies were recorded in any of the study years at a daily temperature colder than 10 ℃.

Influence of various stressors on the expression of core genes of the small interfering RNA pathway in the oriental fruit fly, Bactrocera dorsalis

RNA interference （RNAi）-based technology has emerged as a potential tool for controlling insect pests, however, previous studies found that the efficiency of RNAi in Bactrocera dorsalis was variable. In nature, insects often meet various challenges, such as pathogen infections, extreme temperatures, lack of nutrition and heavy metals. To better understand the association of the stressors with efficiency of RNAi, in the current study we tested the expression of three core genes, dicer2 （Bddcr2）, r2d2 （Bdr2d2） and argonaute2 （Bdago2）, of the small interfering RNA （siRNA） pathway of B. dorsalis upon various stressors. Our results showed that all three genes were upregulated by the infection of invertebrate iridescent virus 6, which suggested a function of the siRNA pathway against viral infection. The loading of FeC13 could also increase the expression of Bddcr2. The treatments ofEscherichia coli, extremely high （40~C） and low （0~C） temperatures, as well as starvation, could negatively influence the expression of Bddcr2 and/or Bdago2. In total, our results showed that various stressors could influence the expression of core components ofB. dorsalis siRNA pathway. This highlights further speculation on the RNAi efficiency upon these stressors. Considering the complexity and variation of RNAi efficiency in different conditions, these results provide initial aspects in possible environmental stressors to influence the activity of the siRNA pathway, but the real impact of RNAi efficiency posed by these stressors requires further studies.