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。     

橘小实蝇的风险分析

从地理评估和管理标准、传人的可能性、传播途径、定殖、定殖后扩散的可能性及橘小实蝇危害性评估等方面对橘小实蝇传人中国风险进行定性分析,筛选了橘小实蝇传人中国的风险因子。根据我国各气候大区的30a（1961—1990）的平均气温、月最高平均气温、月最低平均气温及年降雨量等气候因子和橘小实蝇的发生规律,将我国地域划分为适生区、次适生区、非适生区,明确了南亚热带湿润大区、边缘热带湿润大区、边缘热带亚湿润大区为橘小实蝇的适生区;暖温带湿润大区、北亚热带湿润大区、南亚热带亚湿润大区、高原亚温带湿润大区为橘小实蝇的次适生区。提出了降低橘小实蝇风险的管理措施,为制定检疫政策提供依据。     

橘小实蝇复合体分类学研究进展

橘小实蝇复合体Bactroceradorsaliscomplex是以橘小实蝇命名的实蝇类群,已知75种,广泛分布于亚洲、澳洲及环太平洋周边地区。文章概述基于形态学和遗传学特征基础上的橘小实蝇复合体的分类鉴定及系统发育研究进展,介绍近年来分子生物学技术在橘小实蝇复合体分类及系统发育研究中的应用情况,展望系统发育和行为学研究是橘小实蝇复合体未来研究的重点领域。     

prima蛋白与马拉硫磷混合后点喷对橘小实蝇防治效果的研究

应用prima蛋白加农药马拉硫磷对橘小实蝇进行防治,每8d点喷一次,共喷5次。试验结果表明：prima蛋白与马拉硫磷混合后点喷对橘小实蝇的诱杀效果显著,保果率达到64．71％左右,卡方检验的结果为P=0．027〈0．05,表明点喷prima蛋白与马拉硫磷的混合溶液对柚子的保果效果显著。     

低温对橘小实蝇羽化的影响

【背景】橘小实蝇是重要的检疫性害虫,发生危害严重。有关低温对橘小实蝇成虫羽化的影响尚未见报道。【方法】通过设置不同温度(8、4、0和-4℃)及处理时间(12、18和24 h),研究了橘小实蝇羽化率和羽化时间的变化。【结果】随着温度的降低,橘小实蝇羽化率和羽化时间显著降低和延长。羽化率可从正常温度(28℃)下的100.0%降低至6.7%(0℃);羽化时间可从正常温度(28℃)下的9.0 d延长至12.5 d(0℃)。当温度为-4℃时,橘小实蝇不能正常羽化。【结论与意义】橘小实蝇的耐寒性较差。本研究可为该虫适生区划定提供理论依据。     

几种物质对橘小实蝇的引诱力测定

分别以3种果泥、4种人工合成水果味香精、发酵面粉、蜂蜜、甲基丁香酚、月罗香植物粗提取液为引诱物,测定11种引诱物对橘小实蝇的田间引诱能力。结果表明,水果香精、蜂蜜对橘小实蝇的引诱效果很差;香蕉果泥具有极强的引诱能力,诱虫数达到354只,雌雄比达1∶1;发酵面粉具有较好的诱雌性,诱虫总量达287只,雌雄比为8∶1;甲基丁香酚具有最强的诱雄能力,共引诱了763只橘小实蝇;月罗香粗提取液引诱了429只雄橘小实蝇,具有较强的引诱力。     

橘小实蝇和番石榴实蝇卵的提取物对两种实蝇行为的影响及化学成分分析

[目的]探究橘小实蝇Bactrocera dorsalis和番石榴实蝇Bactrocera correcta卵提取物对2种实蝇雌虫行为的影响,以及2种实蝇卵表面化合物的差异,为2种实蝇的化学生态防治提供理论依据.[方法]利用Y型嗅觉仪测定了橘小实蝇交配雌虫和番石榴实蝇交配雌虫对卵表提取物的行为反应,并利用EthoVision XT软件分析雌虫在Y管内的运动轨迹;利用气相色谱-质谱联用仪(GC-MS)鉴定分析橘小实蝇卵和番石榴实蝇卵的化合物成分,并利用标准曲线法测定了各组分的含量.[结果]橘小实蝇卵提取物对橘小实蝇交配雌虫具有显著引诱作用(x2=9.383,P=-0.002),对番石榴实蝇交配雌虫具有显著驱避作用(x2=6.737,P=0.009),番石榴实蝇卵提取物对番石榴实蝇交配雌虫(x2=4.235,P=0.040)和橘小实蝇交配雌虫都具有引诱作用(x2=5.818,P=0.016).从橘小实蝇卵表提取物中共鉴定出11种化合物,分别是茴香脑、十五烷、十二烷酸、十二酸乙酯、(Z)-11-十四碳烯酸、十四烷酸、十四酸乙酯、(E)-9-十六碳烯酸乙酯、十六烷酸、十八碳烯酸、(Z)-9-十八碳烯酸乙酯.从番石榴实蝇卵表提取物中仅鉴定出7种化合物,而且这7种化合物在橘小实蝇的卵提取物中都存在.仅存在于橘小实蝇卵表的4种化合物是茴香脑、十二烷酸、十二酸乙酯、(Z)-11-十四碳烯酸.在2种实蝇卵提取物的相同成分中,十四烷酸、(E)-9-十六碳烯酸乙酯、十六烷酸在橘小实蝇卵表的含量显著高于番石榴实蝇.[结论]番石榴实蝇具有通过卵表化合物识别同种卵和异种卵的能力,2种实蝇的卵表化合物具有明显的差异.     

橘小实蝇快速检疫鉴定方法

采用形态学观察与PCR技术相结合的方法,针对台湾水果中截获的可疑橘小实蝇Bactroceradorsalis(Hendel)幼虫和蛹进行快速鉴定。采用异硫氰酸胍方法快速提取昆虫基因组DNA,并根据特异性引物,采用PCR技术从截获的可疑橘小实蝇幼虫和蛹样本中均扩增得到224 bp的特异性条带,经测序比较发现,扩增产物序列与橘小实蝇目的基因序列完全相同,由此证明可疑样本为橘小实蝇的幼虫和蛹。该方法解决了橘小实蝇幼虫和蛹等未成熟虫态的快速检疫难题,大大缩短了检测周期,值得口岸检验检疫借鉴应用。     

橘小实蝇与番石榴实蝇卵及蛹的种间竞争

【背景】实蝇的竞争可能发生在生活史的各个阶段,但未见卵和蛹的种间竞争的相关报道。【方法】将橘小实蝇与番石榴实蝇的卵及蛹按照相应的比例或龄期分别进行混合培养,分析2种实蝇的卵或蛹混合后的发育历期和存活率是否受到彼此影响。【结果】橘小实蝇与番石榴实蝇的卵同龄等量混合,对彼此卵的历期和孵化率无影响。当橘小实蝇和番石榴实蝇的蛹为新鲜蛹时,分别与比它们蛹龄大1 d的对方蛹混合,蛹的羽化率分别为(87.67±3.61)%和(84.33±2.56)%,显著小于对照,说明在此混合蛹处理中,后化蛹者的发育可能受到先化蛹者的抑制。【结论与意义】总体上,在卵和蛹期,橘小实蝇与番石榴实蝇未产生竞争作用;但不排除蛹期竞争的可能性,这种竞争可能存在于特定的蛹期。     

橘小实蝇的发生特点与综合防治技术

橘小实蝇(Bactrocera dorsalis)在福建省寄主范围很广,并严重为害寄主。它的主要为害期是每年的4-11月。生产实践中采用综合治理与农业防治技术有效地控制其为害。     

fruitless在桔小实蝇求偶和交配行为中的作用

【目的】fruitless (fru)是昆虫求偶和交配行为中的关键基因,但是其在桔小实蝇Bactrocera dorsalis中的具体功能尚不清楚。本研究旨在明确fru在桔小实蝇求偶交配行为中的作用,进一步明确fru的生物学意义。【方法】根据NCBI上桔小实蝇fru的预测序列设计两端引物,以桔小实蝇羽化10 d已交配的成虫头部cDNA为模板克隆fru基因的全长cDNA,并对其编码蛋白的结构域进行预测。通过实时荧光定量PCR(RT-qPCR)技术检测该基因在桔小实蝇交配前后表达量的变化。待合成fru基因的dsRNA后,注射到羽化9 d后的桔小实蝇雄成虫腹部进行RNAi,分别于注射后48 h和72 h取样,检测fru基因的相对表达量;并于RNAi后72 h进行求偶交配行为观察,测定桔小实蝇的交配频率和交配持续时间。【结果】克隆获得桔小实蝇fru的全长cDNA,长2 865 bp,编码954个氨基酸,预测蛋白分子量104.1 kD,等点为6.01。该基因所编码的蛋白具有特殊结构域,分别为一个BTB (Broad-complex, Tramtrack and Bric-a-bric)结构域和两个锌指(zinc finger)结构域。RT-qPCR结果表明,相比未交配的桔小实蝇成虫,fru基因在雄成虫头部中的表达量在与雌成虫交配后显著上升。相对于空白对照组(注射DEPC水)和阴性对照组(注射dsGFP),注射dsfru的RNAi处理组的雄性求偶时间延长(延长了25~35 min),且交配频率下降(降低了17%~22%),说明fru基因在雄性桔小实蝇的求偶交配行为中发挥着重要作用。【结论】本研究结果为明确fru在非模式昆虫桔小实蝇求偶交配行为中的功能提供参考,并为深入研究桔小实蝇的求偶交配行为和绿色防控提供了理论基础。     

桔小实蝇肽聚糖识别蛋白基因BdPGRP-SB1的克隆及功能鉴定

【目的】为探究肽聚糖识别蛋白(PGRP)基因BdPGRP-SB1在桔小实蝇Bactrocera dorsalis免疫中的作用。【方法】本研究利用PCR克隆桔小实蝇BdPGRP-SB1全长cDNA序列;利用生物信息学软件对该基因核苷酸序列及其编码的氨基酸序列特征进行分析。采用RT-qPCR分析BdPGRP-SB1在桔小实蝇不同发育阶段(卵、幼虫、蛹、成虫)及5日龄成虫不同组织(中肠、马氏管、后肠、脂肪体、卵巢和精巢)中的表达模式;对桔小实蝇5日龄雌成虫分别注射大肠杆菌Escherichia coli 0111:B4肽聚糖(PGN-EB)和金黄色葡萄球菌Staphylococcus aureus肽聚糖(PGN-SA)后检测BdPGRP-SB1表达水平变化。利用RNAi沉默BdPGRP-SB1的表达,测定大肠杆菌和金黄色葡萄球菌诱导后桔小实蝇雌成虫的死亡率及大肠杆菌诱导后抗菌肽(AMP)基因attacin-A, defensin和diptercin表达变化情况。【结果】克隆获得桔小实蝇BdPGRP-SB1的全长cDNA序列(GenBank登录号: MN892482),开放阅读框长558 bp,编码185个氨基酸,其编码蛋白预测分子量为21.45 kD,等电点为8.57。序列分析表明,BdPGRP-SB1无跨膜结构域,具有PGRP保守结构域,前端具有信号肽,为分泌型蛋白;具有Zn²⁺依赖性酰胺酶活性和DAP型肽聚糖识别位点。系统进化分析发现,BdPGRP-SB1与辣椒实蝇B. latifrons的PGRP-SB1亲缘关系最近,氨基酸序列一致性达96%。发育表达模式表明,BdPGRP-SB1在桔小实蝇3日龄幼虫和成虫期高表达;组织表达谱结果显示BdPGRP-SB1在5日龄成虫各组织中均有表达,在脂肪体内表达量最高。PGN-EB和PGN-SA均能诱导桔小实蝇雌成虫体内BdPGRP-SB1表达水平变化。通过RNAi抑制BdPGRP-SB1表达后,注射大肠杆菌导致桔小实蝇雌成虫死亡率显著升高,以及attacin-A, defensin和diptercin表达量显著上调。【结论】结果说明桔小实蝇BdPGRP-SB1参与识别革兰氏阴性细菌,并可能参与桔小实蝇Imd途径调控其免疫反应。     

桔小实蝇抗性品系的微卫星DNA分析

用微卫星标记对桔小实蝇 Bactrocera dorsalis(Hendel) 敏感品系和3个抗性品系（敌百虫抗性品系、高效氯氰菊酯抗性品系和阿维菌素抗性品系）进行了基因组筛查。结果表明,抗性品系显示出微卫星DNA多态性,不同的抗性品系已经产生了不同的遗传变异。该结果有利于指导控制桔小实蝇抗药性产生以及抗药性的田间监控,同时在抗药性产生的遗传学机制研究方面有一定的意义。     

Relationship between female body size and demographic parameters inBactrocera Malaysian A (Diptera: Tephritidae)

The effect of body size, as measured by the head width, of the female Bactrocera sp. Malaysian A (kept separately in sexual pairs) on the demographic parameters was investigated in the laboratory under ambient conditions of 28–30°C, 78–85% RH and natural photoperiod. Body size was shown to influence significantly all the demographic parameters. The expectation of life of females at eclosion from pupae was respectively for head widths of 1.6, 1.8, 1.9, 2.0 and 2.1 mm: 76.2, 73.4, 73.8, 102.4 and 115.2 days. The mean number of eggs laid per female in its life time was respectively: 86.4±48.7, 181.8±56.1, 229.7±72.6, 364.3±69.4 and 477.5±109.3 which was significantly different from one another (F=3.73,P<0.05) especially the two smaller sizes from the two larger sizes. The regression line for total eggs laid (Y) against head width (X) was Y=785.2X−1208.7 (R²=0.35,P<0.001). The net reproductive rate (R₀) was respectively 15.8, 34.0, 43.5, 66.9 and 88.8 eggs, while the intrinsic rate of increase (r) was respectivley 0.0435, 0.0538, 0.0670, 0.0665 and 0.0711. The results confirm that for mass rearing purposes, larger females which produce more offspring are to be preferred.     

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。     

A novel attractant for Anastrepha ludens (Diptera: Tephritidae) from a Concord grape product

An attractant for Mexican fruit fly, Anastrepha ludens (Loew) (Diptera: Tephritidae), was developed from a commercial product called Sabor Uva containing processed Concord grape juice. The principal volatile components of Sabor Uva aroma were identified and an aqueous mixture of 15 components that was gas chromatographically similar to Sabor Uva was prepared. This mixture was equivalent to Sabor Uva in attractiveness by using wind-tunnel bioassays. After deleting chemicals that did not contribute to attractiveness, and increasing the concentrations of the remaining chemicals, the final attractant contained propylene glycol (90,000 ppm, vol/vol), acetic acid (4500), methyl anthranilate (1800), ethyl 2-methylpropionate (670), and one or both of the esters ethyl 3-methylbutyrate (44) and 2-methylbutyl propionate (44), in aqueous solution. This mixture was approximately 1.8X as attractive as Sabor Uva by indirect comparison. Deletion of propylene glycol, acetic acid, methyl anthranilate, or ethyl 2-methylpropionate from the mixture significantly decreased attractiveness. Deletion of either of the other two esters seemed to diminish attractiveness although effects were not statistically significant. Deletion of water from the mixture significantly decreased attractiveness. We conclude that propylene glycol, acetic acid, methyl anthranilate, water, and at least one or as many as all three of the methyl-branched esters are essential for complete attractiveness.     

Effective sampling range of a synthetic protein-based attractant for Ceratitis capitata (Diptera: Tephritidae)

Studies were conducted in Honduras to determine effective sampling range of a female-targeted protein-based synthetic attractant for the Mediterranean fruit fly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Multilure traps were baited with ammonium acetate, putrescine, and trimethylamine lures (three-component attractant) and sampled over eight consecutive weeks. Field design consisted of 38 traps (over 0.5 ha) placed in a combination of standard and high-density grids to facilitate geostatistical analysis, and tests were conducted in coffee (Coffea arabica L.),mango (Mangifera indica L.),and orthanique (Citrus sinensis X Citrus reticulata). Effective sampling range, as determined from the range parameter obtained from experimental variograms that fit a spherical model, was approximately 30 m for flies captured in tests in coffee or mango and approximately 40 m for flies captured in orthanique. For comparison, a release-recapture study was conducted in mango using wild (field-collected) mixed sex C. capitata and an array of 20 baited traps spaced 10-50 m from the release point. Contour analysis was used to document spatial distribution of fly recaptures and to estimate effective sampling range, defined by the area that encompassed 90% of the recaptures. With this approach, effective range of the three-component attractant was estimated to be approximately 28 m, similar to results obtained from variogram analysis. Contour maps indicated that wind direction had a strong influence on sampling range, which was approximately 15 m greater upwind compared with downwind from the release point. Geostatistical analysis of field-captured insects in appropriately designed trapping grids may provide a supplement or alternative to release-recapture studies to estimate sampling ranges for semiochemical-based trapping systems.     

Kairomone attractant for the leafmining fly, Liriomyza bryoniae (Diptera, Agromyzidae)

A field test carried out in an industrial greenhouse in Lithuania revealed the attractiveness of synthetic methyl salicylate (MeSa) towards an economically important leafmining tomato pest, Liriomyza bryoniae (Kaltenbach) (Diptera, Agromyzidae). The behavioural reaction of the flies depended very much on the simultaneous presence of both olfactory and visual stimuli. The attractiveness depended on the colour of a sticky trap: MeSa attracted significantly more flies (ca. 2.2 times) when placed in yellow traps than in aluminium foil colour ones, when catches in such traps were compared to a corresponding control. L. bryoniae is the first species within the Agromyzidae family attracted by MeSa. The attractant was attributed to kairomones, as the compound is known as a plant-produced volatile. MeSa can be an effective extra-tool for increasing the attractiveness of traps. It should be evaluated in future whether such trap/bait combination is effective for the mass trapping of L. bryoniae leafminers in greenhouses (closed area).     

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.