Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) is not invasive through Asia: It's been there all along

Oriental fruit fly, Bactrocera dorsalis (Hendel), is a highly polyphagous fruit fly which, in the last 15 years, has invaded (with or without establishment) Africa, Europe and North America. As a direct result of these invasions, there is increasing research interest in the invasion history and spread patterns of this fly. A statement being repeatedly used in the B. dorsalis invasion literature is that the species was first identified from Taiwan in 1912 and that it subsequently spread through South‐East and South Asia during the 20th century. This assumption is incorrect and stems from: (a) an incomplete knowledge of B. dorsalis taxonomic history; and (b) a confounding of first taxonomic record with first invasion record. Rather than being first detected in Taiwan in 1912, the first record of oriental fruit fly was from “East India” (India orientali) under the synonymous name of Musca ferruginea by Fabricius in 1794, and in the 1910s, it was known not only from Taiwan, but widely across tropical Asia with records from India, Burma, Bengal, Sri Lanka (as Ceylon), Singapore and Indonesia (multiple islands). The taxonomic literature is very clear that oriental fruit fly has not invaded the rest of Asia from Taiwan since 1912, and this error should not continue to be repeated in the literature.     

从东南亚进口水果截获桔小实蝇复合种五近缘种的鉴别(双翅目:实蝇科)

介绍了近年从进口东南亚水果中经常截获的、最具经济重要性的桔小实蝇复合种5个近缘种:杨桃实蝇B.carambolae、桔小实蝇B.dorsalis、芒果实蝇B.occipitalis、木瓜实蝇B.papayae及菲律宾实蝇B.philippinensis,分别记述了翅、胸、足、腹及雌虫产卵器主要鉴别特征,并列出桔小实蝇复合种5近缘种鉴别特征检索表。     

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.     

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。     

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。     

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。     

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。     

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。     

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。     

云南桔小实蝇五个地理种群的遗传分化研究

对云南省瑞丽、景洪、化念、元江、河口的桔小实蝇5个地理种群共27个个体的线粒体细胞色素氧化酶Ⅰ(COⅠ) 基因进行了部分序列测定。序列的碱基变化中转换明显多于颠换、无碱基的插入和缺失。5个地理种群中共有27个多态位点和23种单倍型,其中2种为共享单倍型。桔小实蝇5个地理种群的Fst值在0.0364～0.1364之间(P>0.05), Nm值在3.88～13.25之间。对其所有单倍型聚类分析发现,单倍型在系统树中的分布散乱、混杂,没有显示出明显的地理分布族群。分析认为5个地理种群间已存有一定程度的遗传分化,但分化的程度还比较低。造成5个种群这种遗传分化的因素主要与地理隔离有关,而种群间低程度的遗传 分化估计与桔小实蝇在云南的发展历史及生态适应有关。     

Effect of sex,age and morphological traits on tethered flight of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) at different temperatures

The oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae), is a pest of fruit and vegetable production that has become established in 42 countries in Africa after its first detection in 2003 in Kenya. It is likely that this rapid expansion is partly due to the reported strong capacity for flight by the pest. This study investigated the tethered flight performance of B. dorsalis over a range of constant temperatures in relation to sex and age. Tethered flight of unmated B. dorsalis aged 3, 10 and 21 days was recorded for 1 h using a computerized flight mill at temperatures of 12, 16, 20, 24, 28, 32 and 36 °C. Variations in fly morphology were observed as they aged. Body mass and wing loading increased with age, whereas wing length and wing area reduced as flies aged. Females had slightly larger wings than males but were not significantly heavier. The longest total distance flown by B. dorsalis in 1 h was 1559.58 m. Frequent short, fast flights were recorded at 12 and 36 °C, but long-distance flight was optimal between 20 and 24 °C. Young flies tended to have shorter flight bouts than older flies, which was associated with them flying shorter distances. Heavier flies with greater wing loading flew further than lighter flies. Flight distances recorded on flight mills approximated those recorded in the field, and tethered flight patterns suggest a need to factor temperature into the interpretation of trap captures.     

Mitotic and polytene chromosomes analysis of the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

The Oriental fruit fly, Batrocera dorsalis s.s. (Hendel) is one of the most destructive agricultural pests, belonging to a large group of difficult to distinguish morphologically species, referred as the B. dorsalis complex. We report here a cytogenetic analysis of two laboratory strains of the species and provide a photographic polytene chromosome map from larval salivary glands. The mitotic complement consists of six chromosome pairs including a heteromorphic sex (XX/XY) chromosome pair. Analysis of the polytene complement has shown a total of five polytene chromosomes (10 polytene arms) that correspond to the five autosomes. The most important landmarks of each polytene chromosome and characteristic asynapsis at a specific chromosomal region are presented and discussed. Chromosomal homology between B. dorsalis and Ceratitis capitata has been determined by comparing chromosome banding patterns. The detection of chromosome inversions in both B. dorsalis strains is shown and discussed. Our results show that the polytene maps presented here are suitable for cytogenetic analysis of this species and can be used for comparative studies among species of the Tephritidae family. They also provide a diagnostic tool that could accelerate species identification within the B. dorsalis complex and could shed light on the ongoing speciation in this complex. Polytene chromosome maps can facilitate the development of biological control methods and support the genome mapping project of the species that is currently in progress.     

桔小实蝇肌球蛋白轻链2基因的克隆及表达分析

肌球蛋白轻链在生物的运动过程中具有重要作用, 本研究利用RT-PCR和RACE技术获得了桔小实蝇Bactrocera dorsalis （Hendel）肌球蛋白轻链2 (myosin light chain 2, MLC2)的cDNA全长序列, 命名为BdorMLC2。测序结果表明, BdorMLC2开放阅读框全长669 bp, 编码222个氨基酸残基。在线软件SMART分析显示, BdorMLC2具有2个Ca²⁺结合基序（EFh）结构域, 可以结合Ca²⁺, 属于肌钙蛋白C超家族成员。氨基酸序列比对表明, BdorMLC2与其他昆虫的肌球蛋白具有较高的序列一致性, 其中与黑腹果蝇Drosophila melanogaster MLC2的序列一致性高达93.2%。BdorMLC2在不同组织和时期的荧光定量PCR分析表明, BdorMLC2在桔小实蝇雄虫胸部的含量最高, 在前足、中足、后足和翅中也有较高的表达; BdorMLC2几乎在桔小实蝇发育的各个时期都有表达, 刚羽化成虫的表达量显著高于其他发育阶段。结果提示, BdorMLC2可能与桔小实蝇的肌肉收缩运动密切相关。本研究为深入研究桔小实蝇肌球蛋白的功能提供了理论依据。     

橘小实蝇幼虫液体人工饲料中pH值对其生长发育的影响

本文主要研究了橘小实蝇液体人工幼虫饲料中不同酸度对橘小实蝇生长发育的影响。继代饲养3代的结果表明：饲料酸度对橘小实蝇化蛹率和后代卵孵化率没有影响;对橘小实蝇蛹重、成虫羽化率和产卵量有显著影响。橘小实蝇液体人工幼虫饲料的pH值调节至3.6时仍可以获得质量较高的橘小实蝇。     

桔小实蝇V-ATPase G亚基基因的克隆及组织表达特异性分析

空泡型ATP酶（vacuolar-type H⁺-ATPase, V-ATPase）作为质子泵几乎在所有的真核生物细胞中发挥重要作用。本研究利用RT-PCR和RACE技术获得了桔小实蝇Bactrocera dorsalis (Hendel)V-ATPase G亚基序列全长, 命名为BdorATPG。测序结果表明, BdorATPG阅读框全长354 bp, 编码117个氨基酸。氨基酸序列比对表明, BdorATPG的N端序列与其他物种的ATPG亚基对应区域具有较高的序列一致性。BdorATPG与拟暗果蝇Drosophila pseudoobscura ATPG亚基的氨基酸序列一致性最高, 为88.9%。三维结构模建结果表明, BdorATPG N端（第1~59位氨基酸）序列为α-螺旋结构, 亲水性和疏水性氨基酸在螺旋两侧呈对称分布。BdorATPG在不同组织中的荧光定量PCR分析表明, BdorATPG在各组织中都有表达, 其中在触角中的表达量最高; 在雄虫生殖节中的表达量是雌虫中的6.04倍。结果提示BdorATPG可能在雄虫生殖生理过程中发挥重要作用。     

桔小实蝇和番石榴实蝇幼虫取食阶段的种内竞争

种内竞争是实蝇的重要生态学特性。本文研究比较了近缘种桔小实蝇Bactrocera dorsalis(Hendel)和番石榴实蝇B.correcta(Bezzi)在幼虫人工饲料中种内竞争的密度效应。研究结果显示,桔小实蝇和番石榴实蝇都有密度过低效应,均表现出发育历期较长、蛹较轻和成活率较低;结合蛹重和成活率两个重要参数指标,明确了桔小实蝇和番石榴实蝇的最宜密度均在5-20头虫/10 g饲料;不同生长参数之间的变化均有一定的正负相关性。当种群密度超过最宜密度,桔小实蝇和番石榴实蝇均有明显的种内竞争现象,随着种群密度的升高,种内竞争越激烈。其主要表现为化蛹率和羽化率不断下降、幼虫历期逐渐缩短、蛹历期不断延长和蛹逐渐变小;当100头虫/10 g饲料时,桔小实蝇和番石榴实蝇成活率从86.00%±4.2688%和86.00%±4.1096%分别下降到60.08%±2.5166%和76.33%±2.2333%。桔小实蝇和番石榴实蝇的种内竞争的密度效应极具相似性,推测这两种实蝇有类似的种群调节机制,但番石榴实蝇的幼虫比桔小实蝇的拥有更强的抗拥挤能力。     

基于mtDNA Cytb 的六种果实蝇的分子鉴定

本研究首次对果实蝇属的桔小实蝇Bactrocera dorsalis、瓜实蝇B. cucurbitae、南瓜实蝇B. tau、番石榴实蝇B. correcta、具条实蝇B. scutellata、黑漆实蝇B. scutellaris等6种实蝇mtDNA Cytb基因进行了测序。对这6种实蝇72个个体mtDNA Cytb基因中段420 bp的碱基序列进行分析,得到38种单倍型,发现了116个变异位点,其中30个位点较为稳定。对这6种实蝇与其各自鉴别位点的对应关系研究表明,mtDNA Cytb基因可以作为这6种实蝇种类鉴别的分子标记。     

Purification and biochemical characterization of glutathione S-transferases from four field populations of Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

Glutathione S-transferases (GSTs) are a group of detoxification enzymes that catalyze the nucleophilic addition of glutathione to a wide variety of endogenous and xenobiotic compounds. In this study, GSTs were purified from four field populations of Bactrocera dorsalis with different insecticide susceptibilities by glutathione-agarose affinity chromatography. The populations were collected from Dongguan (DG) and Guangzhou (GZ) of the Guangdong Province, Haikou of the Hainan province (HN), and Kunming of the Yunnan province (YN), China. Differences in GST characteristics among the four populations were studied using purified enzyme samples through comparative SDS-PAGE, kinetic, and inhibition experiments. The specific activities of the purified enzymes were similar, but the purification yield of the GZ population (31.54%) was the lowest. SDS-PAGE analysis showed only one band at approximately 23 kDa for these four populations. Kinetic analyses showed that the affinities of the purified GSTs from the GZ and YN populations for 1-chloro-2.4-dinitrobenzene (CDNB) were much higher than those of GSTs from the other two populations, whereas the HN population had the highest catalytic capability in terms of V(max) value. The optimum temperature for CDNB conjugation was 37 °C and the optimum pH was 7.5 in all four populations. Inhibition kinetics showed that ethacrynic acid, diethyl maleate, tetraethylthiuram disulfide, curcumin, bromosulfalein, and β-cypermethrin had excellent inhibitory effects on GSTs in the four populations of B. dorsalis, but the low inhibitory effects of malathion and avermectin did not differ between populations. These results suggest that GSTs may have a role in detoxification of β-cypermethrin in B. dorsalis.     

桔小实蝇自然种群蛹和越冬成虫的耐寒性

本文通过过冷却点和低温耐寒能力的测定,研究了我国桔小实蝇Boctrocera dorsalis (Hendel)不同季节种群和地理种群蛹的耐寒性规律。结果表明：桔小实蝇自然种群蛹的低温存活力有明显的季节规律,冬前种群的耐寒性显著强于夏季种群。如在0℃低温下暴露2天,夏季种群基本全部死亡,而冬前种群存活率仍为61.4%。2005-2006年不同月份桔小实蝇1日龄蛹过冷却点差异明显,以2月份均值最高,为-9.26℃,8月份最低,为-15.20℃;且2月份桔小实蝇蛹个体过冷却点值出现明显的分化现象,分别在-15℃左右和-6℃左右出现两个明显的聚集区。所研究的5个桔小实蝇地理区域种群没有出现明显的耐寒性分化。结合本文结果,文中还就桔小实蝇自然种群耐寒性影响因素进行了讨论。