Global distribution and invasion pattern of oriental fruit fly,Bactrocera dorsalis (Diptera: Tephritidae)

Since the start of the 20th century, many invasive alien species (IAS) have spread rapidly around the world, causing serious threats to economies, societies and the environment. Bactrocera dorsalis (Hendel) (Diptera: Tephritidae) is an important quarantine insect species in many countries that spread around the world over the last century. This review collected information on the distribution of B. dorsalis to explore the patterns of its invasion expansion. We found B. dorsalis to be distributed in 75 countries (comprised of 124 geographical distribution regions: provinces or states) in Asia, Africa, North America, South America and Oceania up to 2017. Asia and Africa were the most represented regions, accounting for 86.3% of the total number of countries. From 1910 to 1990, B. dorsalis was only found in five countries, but in the last three decades, it has experienced a sharp increase in its rate of spread, invading 70 more countries. Global temperature anomaly has significantly positive correlation with the spread of B. dorsalis. The results of this review provide a theoretical basis for understanding and predicting the continued spread of B. dorsalis under global changes.     

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。     

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.     

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.     

桔小实蝇雄成虫联系性嗅觉学习

【目的】为了探究桔小实蝇 Bactrocera dorsalis (Hendel)雄成虫的嗅觉学习能力。【方法】本研究采用经典性嗅觉条件反射训练法（classical olfactory conditioning）在室内对固定的羽化后14-17日龄的桔小实蝇雄成虫进行气味与食物的联合学习训练, 即薄荷精油和10%蔗糖溶液联合的奖赏性训练（appetitive conditioning）以及甲基丁香酚(methyl eugenol, ME)和饱和盐溶液联合的惩罚性训练（aversive conditioning）,并以伸喙反射行为（proboscis extension reflex, PER）作为学习与否的判定标准。【结果】经过奖赏性训练后,桔小实蝇雄成虫对薄荷精油的伸喙反射率可从0%增加至68%;而经过惩罚性训练后,桔小实蝇对甲基丁香酚的伸喙反射率可从100%降低至36.54%,且这种伸喙反射率的变化是通过气味条件刺激（conditioned stimulus）和食物非条件刺激（unconditioned stimulus）的对称性联合而产生的。【结论】结果表明,桔小实蝇雄性成虫具有较强的联系性嗅觉学习能力,并且两种刺激的联合是形成学习记忆的必要条件。     

The complete nucleotide sequence of the mitochondrial genome of the oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae)

The complete mitochondrial genome of the oriental fruit fly Bactrocera dorsalis s.s. has been sequenced, and is here described and compared with the homologous sequences of Bactrocera oleae and Ceratitis capitata. The genome is a circular molecule of 15,915 bp, and encodes the set of 37 genes generally found in animal mitochondrial genomes. The structure and organization of the molecule is typical and similar to the two closely related species B. oleae and C. capitata, although it presents an interesting case of putative intra-molecular recombination. The relevance of the growing comparative dataset of tephritid complete mitochondrial genomes is discussed in relation to the possibility to develop robust assays for species discrimination in quarantine and agricultural monitoring practices, as well as basic phylogeography/population genetic studies.     

矿物油乳剂拒避橘小实蝇的有效组分分析

高品质的矿物油乳剂可用于有机食品生产,在害虫综合治理中有良好的应用前景。矿物油是包含多种化学成分的混合物,其组成成分和比例决定其理化性质并关系到其防治病虫害的效果。已有研究报道表明,矿物油乳剂对许多害虫有拒避作用,但其机理及相关性组成成分尚未研究清楚。本实验测定了4种矿物油乳剂即SK绿颖农用喷洒油、加德士-路易夏用油、安波尔喷洒油、法夏乐石蜡油（分别简称为SK,Caltex,Ampol和Citrole油）和Caltex油＜120℃,＞120℃,＜105℃,105~120℃及SK油＜110℃、＞110℃等6种分子蒸馏组分的正构烷烃碳数当量nCy值、碳数分布值及其对橘小实蝇Bactrocera dorsalis的产卵拒避效果。室内生物测定结果显示,橘小实蝇在SK,Caltex,Ampol和Citrole油0.5%水乳液及清水浸蘸处理后的香蕉上平均产卵孔数分别为17.850±1.282,3.400±0.630,15.850±1.439,20.650±1.212和19.050±1.155,在同样方法处理的阳桃上平均产卵孔数分别为15.500±3.969,3.000±0.707,13.750±4.131,17.250±4.385和62.000±4.708。橘小实蝇在6种分子蒸馏组分0.5%水乳液处理后的香蕉上的平均产卵孔数分别为4.400±1.166,8.200±0.583,5.400±1.720,2.400±1.077,4.800±1.594和12.200±2.63,显示不同矿物油的拒避效果差异显著（P<0.05）,拒避效果最好的矿物油的正构烷烃碳数当量nCy值在nC23左右.矿物油对橘小实蝇的产卵拒避效果与其相应的碳数分布值的Spearman等级相关性分析显示,其拒避效果与矿物油中的C23组分的含量呈显著正相关关系（P<0.05）。这可为进一步分析矿物油乳剂拒避害虫的作用成分和探索其机理提供参考。     

桔小实蝇对敌百虫抗性稳定性及再增长趋势

通过室内试验,研究了用LC₅₀及LC₉₀两种浓度敌百虫汰选14代建立的不同程度高抗性水平的桔小实蝇Bactrocera dorsalis抗性品系TrR₅₀和TrR₉₀在停止使用药剂后的抗性衰退规律,以及抗性衰退到原有水平的1/3时再用药汰选的抗性再增长规律。结果表明：两抗性品系对敌百虫抗性均不稳定,但抗性衰退速率不同,TrR₉₀完全隔离药剂4代后,抗性已衰退至原有水平的1/3,而TrR₅₀完全隔离药剂7代后,抗性才衰退至原有水平的1/3,但随后两品系抗性衰退均减缓,到19代后抗性均还处于低水平抗性阶段,无法衰退至敏感水平。通过对其抗性衰退趋势进行方程拟合,结果表明两品系抗性衰退均符合S型曲线模型。抗性再增长试验结果表明：桔小实蝇两抗性品系对敌百虫抗性再增长趋势不同,TrR₅₀继续使用药剂汰选,经过6代选育,抗性迅速上升,接近原有的抗性水平,随后保持平稳增长; 而TrR₉₀继续使用药剂汰选,经过9代选育,抗性才迅速上升,经过12代选育,抗性才接近原有的抗性水平。通过对两品系抗性再增长趋势进行方程拟合,结果表明两品系抗性再增长趋势不同,TrR₅₀品系抗性再增长趋势符合逻辑斯蒂模型,而TrR₉₀品系则符合二次曲线模型。     

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 ℃.     

Linkage analysis of five new genetic markers of the oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae).

Five new autosomal recessive genes are described in the oriental fruit fly, Bactrocera dorsalis (Hendel). These genetic markers are associated into three linkage groups. The matte (mt) gene is linked to the previously described mandarin red (ma) gene, and the white puparium (wp) gene is linked to the white eye (we) and amethyst (am) loci. The third designated linkage group has the yellow eye (ye) marker. The we/we homozygote is epistatic to ye/ye, and each is epistatic to am/am and ma/ma.     

基于线粒体COI基因的桔小实蝇种群遗传分化研究

【目的】推测桔小实蝇Bactrocera dorsalis在中国的扩散路径和新发生地区的入侵来源。【方法】本研究测序获得来自中国、泰国、日本、老挝、孟加拉国和美国等地31个种群的192头桔小实蝇个体的COI序列（1 496 bp,占COI基因全长97.3%）,并以软件DnaSP 5.0, MEGA 6.0和Arlequin 3.5等完成各种群的遗传多样性、种群间的遗传分化以及单倍型分析。【结果】所测31个桔小实蝇种群总体表现出较高水平的核苷酸多样性（0.00663）和高水平的单倍型多样性（0.98069）。以F-统计法度量种群间遗传分化程度, 结果显示中国桔小实蝇地理种群间遗传分化较弱, 中国种群与泰国、日本、老挝、孟加拉国、美国种群间的遗传分化程度不同, 中国种群与美国种群及日本种群的遗传分化最大。而Mantel检验发现,中国、泰国、日本、老挝、孟加拉国和美国种群间的遗传分化与空间距离有关（R=0.670, P<0.0001）, 中国种群间的遗传分化不是地理隔离所造成的（R=0.038, P=0.534）。中国种群与泰国、日本、老挝、孟加拉国和美国种群间不存在共享单倍型。根据种群系统发育树,可将中国原发生地区种群划分为西南、东南两大分支。中性检验和错配分析结果均表明桔小实蝇曾存在大规模的扩张。【结论】桔小实蝇以东南地区和西南地区为源头向中国内陆扩散,其中广东、福建、广西和贵州种群为中国内陆种群较为有影响力的源头。根据遗传多样性、遗传分化、单倍型分析,推测新发生地区桔小实蝇的来源, 例如安徽合肥桔小实蝇种群主要来源于福建长乐、广东珠海和上海。     

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。     

A novel point mutation of acetylcholinesterase in a trichlorfon-resistant strain of the oriental fruit fly Bactrocera dorsalis (Diptera: Tephritidae)

Acetylcholinesterase (AChE) is the target enzyme of organophosphorus and carbamate insecticides. We applied trichlorfon to select resistant strains of Bactrocera dorsalis Hendel in the laboratory. Two trichlorfon-resistant strains, the Tri-R₁ strain with 18.23-fold resistance and the Tri-R₂ strain with 69.5-fold resistance, were obtained. Three known mutations, I159V, G433S and Q588R were identified in AChE of two resistant strains, and a novel mutation, G365A, was identified in the more resistant Tri-R₂ strain. The modeled 3-D-structure of AChE showed that G365A and G433S are closely adjacent in the gorge above the catalytic site S235. Mutations of G365A and G433S resulted in a steric hindrance by stronger Van der Waals force between two sites. Such a minor structural change might block insecticides from squeezing through the gorge to reach the active site, but not the natural substrate. Compared with the susceptible strain, the AChE activity of the Tri-R₁ strain and the Tri-R₂ strain was 0.87- and 0.67-fold, the K _m value of the Tri-R₁ strain and the Tri-R₂ strain was 0.11- and 0.10-fold, the V _max value of two resistant strains was 0.26- and 0.15-fold, whereas, the I ₅₀ to trichlorfon significantly increased by 9.07- and 13.19-fold. These results suggested that the novel point mutation G365A of AChE might be involved in increasing resistance to trichlorfon in the resistant strain of oriental fruit fly.     

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。     

Phosphoglucomutase polymorphism in the oriental fruit fly, Dacus dorsalis (Diptera, Tephritidae) from Peninsular Malaysia

Seven natural populations of Dacus dorsalis were analysed for phosphoglucomutase by means of horizontal starch-gel electrophoresis. The electrophoretic phenotypes were governed by four codominant Pgm alleles. The commonest allele in all the seven population samples was PgmB which encoded an electrophoretic band with intermediate mobility. The distributions of PGM phenotype were in accordance with Hardy-Weinberg expectations. There was geographic variation in the distribution of Pgm alleles.     

cDNA isolation, expression, and hormonal regulation of yolk protein genes in the oriental fruit fly, Bactrocera dorsalis (Hendel) (Diptera: Tephritidae)

Yolk protein (YP) or vitellogenin (Vg), the main component of yolk, is the key nutrient for embryonic development. YPs, encoded from uncleaved genes existing mainly in cyclorraphan flies, are different from VGs that are present in most non-cyclorraphan dipterans and other insects. In this study, cDNAs of two YPs, namely Bdyp1 and Bdyp2 (GenBank accession Nos. AF368053 and AF368054), were isolated in the oriental fruit fly, Bactrocera dorsalis (Hendel). RT-PCR analysis revealed that Bdyp1 and 2 are expressed in the fat body and ovary during egg development. However, the expression profiles of Bdyp1 and 2 in the fat body are different, indicating that divergent mechanisms might exist in the regulation of these two genes. Twenty-hydroxyecdysone (20E) plays a major role in promoting Bdyp1 expression, yet the expression of Bdyp2 exhibits a greater response to juvenile hormone (JH) in fat body in vitro. Unexpectedly, 20E-induced expression of both Bdyp1 and 2 is suppressed by JH prior to 20E treatment of in vitro fat body; conversely, it is enhanced by the addition of JH following 20E treatment.     

植物果实、颜色和形状对橘小实蝇产卵选择的影响

橘小实蝇Bactrocera dorsalis(Hendel)是近几年入侵江苏南部地区,严重危害当地水果、蔬菜的重要害虫。为深入了解其在该地区果树、蔬菜上的产卵特性,进行植物果实、颜色和形状对其产卵选择影响的研究。实验室中"纸杯法"的产卵选择试验结果表明,橘小实蝇对不同水果的产卵偏好性由高到低依次为:柑橘、香蕉、芒果、枣、石榴、苹果、桃子、梨、李、葡萄、柿;对不同蔬菜的产卵偏好性由高到低依次为:丝瓜、豇豆、苦瓜、黄瓜、茄子、南瓜、青椒、番茄。对不同寄主颜色和形状的产卵选择特点为,雌虫常选择桔黄色、绿色,以及圆球形的寄主产卵,而较少选择黑色、方形的寄主产卵。从试验结果可以看出,寄主果实散发出的气味,及其颜色和形状对橘小实蝇的产卵选择有影响。