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.     

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.     

Feeding behaviour of Bactrocera cacuminata (Hering) (Diptera: Tephritidae) on methyl eugenol: a laboratory assay

Abstract Many dacine (Diptera: Tephritidae) species are attracted to one of two chemical substances, 4-allyl-1,2-dimethoxybenzene (methyl eugenol (ME)) or 4-( p -hydroxyphenyl)-2-butanone (cue lure). Despite the fact that these chemicals or analogs occur naturally, their significance in the biology/ecology of the Dacinae has seldom been examined. In this study, we examine the patterns in feeding behaviour of Bactrocera cacuminata (Hering), a monophagous non-pest fruit fly, on ME. Based on a laboratory assay, we found that males of this species fed on ME multiple times within a single day and on multiple days. The pattern of repeat feeding was not related to time since previous feeding or duration of prior feeding. Our results contrast those obtained on a similar study on a related polyphagous species ( Bactrocera dorsalis (Hendel)). We discuss the implications of our findings with a view to explaining the functional significance of dacine response to these plant-derived chemicals.     

寄主对桔小实蝇耐寒性的影响

为了研究寄主营养对桔小实蝇耐寒性的作用,测定了以15种果蔬饲养的桔小实蝇1日龄蛹的过冷却点(supercooling points,SCP); 再选取南瓜、西红柿、柑桔、番石榴和杨桃等5种果蔬,测定了桔小实蝇3龄老熟幼虫、1日龄蛹、3日龄蛹、5日龄蛹、7日龄蛹和雌雄成虫的过冷却点,并观察了1日龄蛹的低温存活力。结果表明：（1）15种果蔬饲养所得的桔小实蝇1日龄蛹SCP均值在-11.03℃～-13.17℃,不同寄主发育的桔小实蝇SCP值存在显著性差异,其中以取食蒲桃的最高,为-11.03℃,取食苦瓜的最低,为-13.17℃。（2）5种果蔬饲养所得的桔小实蝇各虫态的SCP均值存在极显著差异(F_(4,863)＝35.6,P<0.01); 同一寄主上的桔小实蝇不同虫态之间SCP均值也达到极显著性差异(F_(6,863)＝392.9,P<0.01); 且寄主和发育龄期之间存在着极显著的交互作用(F_(24,863)＝9.4,P<0.01)。（3）桔小实蝇各发育阶段,SCP值表现一定变化： 老熟幼虫发育至1日龄蛹,SCP值变化不大; 蛹发育至3、5和7天过冷却能力明显增强,降至-20℃左右,但他们之间没有明显区别; 羽化后3～5天的成虫SCP值又升高至-10℃左右。老熟幼虫、1日龄蛹和2～3日龄成虫与3日龄、5日龄和7日龄蛹的SCP值之间有显著性差异。（4）将5种果蔬饲养所得的桔小实蝇1日龄蛹置于6℃和-3℃下进行较长时间(1～8天)和较短时间(1～8 h)的低温处理,发现番石榴、杨桃和南瓜发育的蛹经低温处理后的校正羽化率较西红柿和柑桔发育的蛹高; 同样在0℃、3℃、6℃和9℃处理(2天)的实验中,得出相似的结果。因此,本实验结果表明桔小实蝇幼虫由于生活寄主的不同使得其下一代蛹的耐寒性产生了差异,引起其差异的原因值得进一步研究。     

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.     

温度对桔小实蝇种群发育的影响

采用室内人工饲养,研究了温度对外来入侵无锡地区的桔小实蝇(Bactrocera dorsalis)发育的影响。结果表明：17℃～33℃桔小实蝇各虫态发育速率逐渐增加;通过直线回归模型与直接最优法,得出桔小实蝇从卵期到成虫期的发育起点温度为9.9 ℃,卵、幼虫和蛹发育所需温度分别为10.5 ℃、9.6 ℃和10.3 ℃;卵期发育到成虫期的有效积温为381.0 d·℃,卵期、幼虫期和蛹期分别为24.1、178.6和176.7 d·℃。此外,温度影响桔小实蝇各虫态的存活率;25 ℃下卵期的存活率最高,而17 ℃下蛹期的存活率最低。温度对桔小实蝇雌雄性比也有一定影响,以25 ℃下雌虫比率最高。     

Captures of pest fruit flies (Diptera: Tephritidae) and nontarget insects in BioLure and torula yeast traps in Hawaii

MultiLure traps were deployed in a Hawaiian orchard to compare the attraction of economically important fruit flies and nontarget insects to the three-component BioLure and torula yeast food lures. Either water or a 20% propylene glycol solution was used to dissolve the torula yeast or as capture fluid in BioLure traps. Torula yeast in water was more attractive than BioLure for male and female Bactrocera cucurbitae (Coquillett) and Bactrocera dorsalis (Hendel) and as attractive for Ceratitis capitata (Wiedemann), and the addition of propylene glycol significantly inhibited the attractiveness of torula yeast. The known synergistic effect of propylene glycol with BioLure, resulting in increased captures of Anastrepha flies, was not observed with Bactrocera. Nontarget Drosophilidae, Neriidae, Phoridae, Calliphoridae, Sarcophagidae, and Muscidae were more strongly attracted to BioLure, and both lures collected Chloropidae equally. As with fruit flies, propylene glycol in torula yeast significantly decreased nontarget captures. The results therefore suggest that torula yeast in water is a more effective attractant than BioLure for pest Bactrocera while minimizing nontarget captures.     

Nontarget insects captured in fruit fly (Diptera: Tephritidae) surveillance traps

Traps baited with synthetic lures (ammonium acetate and putrescine) captured as many Mexican fruit flies as the traditional torula yeast/borax slurry, but with far fewer (ratio 5:1) nontarget insects. Ninety percent of the nontarget insects were dipterans. Consequently, neither trap is efficacious against other citrus pests, which are mainly Hemiptera or Lepidoptera. Although the nontarget catch is sometimes referred to as "trash," many nontarget insects are beneficials, including predators and parasites (especially tachinids). The traps with synthetic lures killed fewer of these beneficials by a ratio of 4:1 compared with the yeast-baited traps. Certain taxa, notably the chrysopids and halictid bees, exhibited a somewhat greater preference (10 and 50%, respectively) for the synthetic lures. Overall, with regard to the deployment of the newer baits, the threat to predators, parasites, and pollinators was found to be negligible, and certainly much less than that posed by the traditional traps.     

桔小实蝇超氧化物歧化酶基因SOD3的克隆及表达分析

采用反转录聚合酶链式反应(RT-PCR)和快速扩增c DNA末端(RACE)技术克隆桔小实蝇SOD3基因,并命名为Bdor SOD3。Bdor SOD3阅读框全长531 bp,编码176个氨基酸,第1-20位氨基酸为其信号肽区域;该蛋白序列与桔小实蝇的另外一种SOD蛋白AGE89778.1序列的一致性最高,达98.7%;采用Swiss-model在线软件模拟构建Bdor SOD3蛋白的三维结构;采用半定量PCR方法,研究Bdor SOD3基因大肠杆菌诱导后的表达情况,结果表明,Bdor SOD3在处理与对照的24 h、48 h都有表达,但Bdor SOD3在处理后48 h表达量明显升高,结果暗示Bdor SOD3与桔小实蝇蛹对大肠杆菌的免疫机制有关。     

桔小实蝇不同发育阶段过冷却点的测定

对桔小实蝇Bactrocera dorsalis (Hendel)不同发育阶段的过冷却点进行了测定。结果表明：同一虫期个体间的过冷却点出现不同程度的变异,但均服从正态分布。不同虫期的过冷却点差异显著,其中蛹的过冷却点最低（-12.2℃～-15.0℃）。老熟幼虫的过冷却点为-8.1℃。成虫的过冷却点最低值为7日龄雄虫（-10.5℃）和雌虫（-10.1℃）,最高值为60日龄雄虫（-5.9℃）和雌虫（-6.4℃）,但同一发育时期的雌、雄成虫之间的过冷却点没有差异。测定结果提示蛹期最有可能是该虫在温带地区越冬的虫态。     

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.     

应用微卫星标记分析不同桔小实蝇种群的遗传多样性

为探究不同桔小实蝇Bactrocera dorsalis (Hendel)地理种群的遗传变异、入侵来源和扩散情况,利用13对引物对中国南方10省区、泰国、夏威夷、菲律宾和老挝的30个桔小实蝇种群共180个个体的遗传多样性水平进行了研究。 Popgene32和NTSYS-pc2.10e软件分析结果表明：30个不同桔小实蝇种群的遗传相似度在0.3599~0.9153范围内。种群的Nei氏基因多样性指数平均为0.6464±0.1026,Shannon信息指数I平均为1.2845±0.2632, 提示桔小实蝇种群具有较丰富的遗传多样性。UPGMA聚类分析显示,福建地区和海南地区分别独立一支,广东地区和台湾地区种群聚成一支,而广西、泰国、湖南、云南、老挝、四川、 重庆和贵州地区聚为一大支系。据此提出泰国种群和老挝种群是最早入侵我国的种群,云南地区是最早的入侵地,广西地区可能为又一较早入侵地。     

Comparison of methyl eugenol metabolites, mitochondrial COI, and rDNA sequences of Bactrocera philippinensis (Diptera: Tephritidae) with those of three other major pest species within the dorsalis complex

Males of the oriental fruit fly, Bactrocera dorsalis (Hendel) and some of its sibling species have strong affinity for methyl eugenol (ME). Methyl eugenol ingested by male flies is biotransformed in the crop to two ME metabolites that eventually accumulate in the rectal gland, which is known to serve as a reservoir for B. dorsalis sex pheromones. When fed with ME, males of laboratory and wild B. philippinensis Drew and Hancock selectively accumulated two metabolites, 2-allyl-4,5-dimethoxyphenol and (E)-coniferyl alcohol, in the rectal gland, as was seen for B. dorsalis sensu stricto, B. invadens Drew, Tsuruta and White, and B. papayae Drew and Hancock. Phylogenetic analysis of COI and rDNA sequence data of these four taxa also revealed a close relationship among B. philippinensis, B. dorsalis s.s., B. invadens, and B. papayae (all four are members of the dorsalis species complex). This result corroborates pheromone analysis. The usefulness of pheromonal analysis as a chemotaxonomy tool to complement molecular and other analysis in differentiation of closely related sibling species within the Bactrocera dorsalis complex, for which use of morphological characters had been inadequate, is highlighted.     

橘小实蝇和番石榴实蝇重复利用寄主的产卵选择性

检疫性害虫橘小实蝇Bactrocera dorsalis和番石榴实蝇Bactrocera correcta是近缘种。本研究显示,两种实蝇对已被产卵寄主的识别与产卵选择性极相似。当寄主中至少含有600粒新鲜卵时,橘小实蝇或番石榴实蝇的雌成虫在其上继续产卵的数量才被抑制,而对同种实蝇卵和异种实蝇卵的反应几乎没有差异;两种实蝇在已被产卵的寄主中再次产卵,并没有刻意回避原产卵孔;寄主中的卵孵化为幼虫对后续实蝇的产卵行为产生了显著的抑制作用,随着幼虫数量的增多与龄期的增长,便不再有雌蝇选择该寄主产卵。因此,寄主中幼虫的存在是驱避橘小实蝇和番石榴实蝇在其上继续产卵的重要因素。两种实蝇幼虫寄主标记信息素的鉴定与利用将是今后研究的重点,利用其进行有害实蝇的种群控制将有很大的应用前景。     

延迟交配对橘小实蝇繁殖适度的影响

橘小实蝇Bactrocera dorsalis Hendel是一种世界性的入侵害虫,严重为害多种经济果蔬。本文通过响应面分析方法,研究了延迟交配对橘小实蝇交配率、产卵量、孵化率和寿命的影响。结果表明,交配率随雌虫交配日龄的延迟先增后减,雌、雄成虫均为35日龄处理组中的交配率最高,为81.00%±2.00%;而雄虫交配日龄的延迟则对其卵孵化率有明显的负面作用,在56日龄雄虫与35日龄雌虫处理组中,其卵孵化率最低,仅为27.56%±4.55%。雌成虫产卵量和寿命受到雌、雄虫双方交配日龄的的影响,产卵量（y）与雄虫日龄（x1）和雌虫日龄（x2）的回归方程为y=514.36+3.08x1-11.05x2,雌成虫寿命（y）与雄虫日龄（x1）和雌虫日龄（x2）的回归方程为y=35.85+0.23x1+0.40x2。研究结果为田间使用性信息素迷向法防治橘小实蝇提供了理论依据。     

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.     

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.     

Movement of sterile male Bactrocera cucurbitae (Diptera: Tephritidae) in a Hawaiian agroecosystem

The melon fly, Bactrocera cucurbitae Coquillett, invaded the Hawaiian Island chain in 1895. In 1999, a program sponsored by the USDA-ARS to control melon fly and other tephritid pests in Hawaii over a wide area was initiated on the islands of Hawaii, Maui, and Oahu. To control these flies in an areawide setting, understanding how flies move within the landscape is important. To explore the movement of this fly, we examined the movement of marked, male, sterile, laboratory-reared B. cucurbitae on the island of Hawaii in an agricultural setting. Two releases of dyed, sterile flies consisting of approximately 15,000 flies, were released 6 wk apart. Released flies were trapped back by using Moroccan traps baited with a male attractant. These two releases suggest that in the Hawaiian agricultural areas where the areawide control is being sought, melon flies do not move extensively when there are abundant larval host and adult roosting sites. Over the course of this study, only one fly made it the maximum distance that we could detect fly movement (approximately 2,000 m in 2 wk). From these data, it seems that the flies dispersed throughout the study area but then moved very little thereafter. This is very apparent in the second release where the recovery rate after the second week was still fairly high, suggesting that if there are plenty of host fields and roosting sites the flies are unlikely to move.     

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。