烟粉虱生物型的监测及其遗传结构研究

烟粉虱Bemisia tabaci(Gennadius)是一种重要的农业害虫并包括许多生物型,其中B型和Q型是入侵性较强的2种生物型。文章着重介绍近年来在烟粉虱生物型的监测及其遗传结构方面的研究进展。B型烟粉虱和Q型烟粉虱这2个生物型均已入侵我国,其中多数地区烟粉虱是B型烟粉虱,局部地区有Q型烟粉虱并呈现不断蔓延趋势。微卫星(SSR)分子标记分析结果表明我国B型烟粉虱的入侵来源具有多元化,而云南地区Q型烟粉虱来源比较单一。化学农药的使用能够影响室内种群的遗传结构,降低种群的遗传多样性。基于RAPD、ISSR分子标记的分析结果表明,Q型烟粉虱种群各项遗传多样性指数均比B型烟粉虱的高。今后加强烟粉虱入侵生物型的遗传结构及其种群动态关系等方面的研究,对于揭示烟粉虱的灾变机制及其控制具有重要的意义。     

基于微卫星标记对中国Q型烟粉虱早期入侵种群与B型烟粉虱种群的遗传结构分析（英文）

2003年首次在云南昆明发现Q型烟粉虱Bemisia tabaci (Gennadius)传入中国。随后几年时间内, 它在许多省份逐年取代了B型烟粉虱种群。2008年后,Q型烟粉虱基本上成为了中国多数省份农区的优势生物型。为了进一步揭示Q型烟粉虱在中国快速扩散以及取代B型烟粉虱的遗传学基础, 本研究利用11个微卫星位点分析并比较了2003年中国云南昆明Q型烟粉虱入侵种群及其他地点的11个B型入侵种群, 西班牙2个Q型土著种群, 以色列1个Q型入侵种群, 以色列1个B型土著种群, 以及西班牙、 美国与澳大利亚的5个B型入侵种群的遗传结构。结果表明, 中国Q型烟粉虱早期种群（云南昆明种群）可能来自于西部地中海地区。中国B型烟粉虱种群遗传多样性高于西班牙、 澳大利亚、 美国B型种群, 中国B型可能存在多次传入或某个混合种群的再次传入。相对于原产地种群, 中国Q型烟粉虱早期入侵种群与B型烟粉虱种群遗传多样性并没有明显降低, 表明Q型与B型烟粉虱种群可能经历了较小的瓶颈效应或奠基者效应。中国Q型烟粉虱早期入侵种群遗传多样性高于B型烟粉虱种群, Q型烟粉虱这种较高的遗传多样性可能为其较强的生态适应性提供了遗传基础, 有利于Q型烟粉虱在新的环境下快速扩散并取代B型烟粉虱。     

北京和湖南烟粉虱生物型及其抗药性监测

烟粉虱是蔬菜等园艺作物上为害严重、最难以防治的害虫之一,生产上防治该虫主要还是依靠化学药剂。本研究鉴定了北京、湖南地区设施蔬菜上烟粉虱的生物型,并监测了其对6种不同杀虫剂的抗药性。结果表明,北京、湖南地区蔬菜烟粉虱生物型分别是Q型和B型。两地烟粉虱对阿维菌素尚处于敏感状态;对烯啶虫胺及其他烟碱类杀虫剂噻虫嗪和吡虫啉的抗药性达中抗到高抗水平,湖南地区抗性水平比北京地区更高,抗性倍数最高达到71.58倍;传统杀虫剂毒死蜱和联苯菊酯对湖南、北京两地烟粉虱的毒杀活性差。     

烟粉虱B型和Q型群体遗传结构的RAPD分析

近20年来,烟粉虱B型传入世界各地并暴发成灾,成为一种重要的农业入侵害虫; 烟粉虱Q型则是近几年引起人们高度重视的一种新的入侵生物型,目前已传入许多国家并造成一定危害。本文利用RAPD分子标记对烟粉虱B型和Q型不同地理种群的遗传结构进行了分析。结果表明：（1）引物H16对烟粉虱B型不同种群扩增的特异带,能有效区分烟粉虱B型和Q型、浙江非B/Q型种群;（2）烟粉虱Q型种群各项遗传多样性指数均比烟粉虱B型的要高;（3）我国烟粉虱Q型来自伊比利亚半岛的可能性比来自中东地区的可能性要大。另外,聚类分析结果提示,RAPD分子标记能有效地区分烟粉虱不同生物型,但可能不适用于生物型之间亲缘关系分析。     

江苏Q型烟粉虱抗性监测及其靶标基因克隆

应用成虫浸叶生测法研究了江苏省扬州、无锡和东台3个地区Q型烟粉虱Bemisia tabaci(Gennadius)田间种群对5种杀虫剂的抗药性。结果表明,与相对敏感种群相比,Q型烟粉虱田间种群已对氯氰菊酯和吡虫啉产生了低到中等水平抗性,对阿维菌素仍然表现敏感。进一步通过RCR扩增获得了长度分别为287bp和184bp的烟粉虱乙酰胆碱酯酶ace1和para-同源钠离子通道基因片断。序列分析表明,江苏Q型烟粉虱存在与有机磷抗性相关的乙酰胆碱酯酶F331W突变和与拟除虫菊酯抗性相关的钠离子通道L925I和T929V突变。     

福建省烟粉虱田间种群抗药性发展及其影响因素

采用成虫浸叶生测法对福建省不同地区烟粉虱田间种群的抗药性发展进行监测.结果表明： 福建各地烟粉虱田间种群对氯氟氰菊酯、甲氰菊酯、毒死蜱仍保持较高水平抗性,对灭多威的抗性水平较低,对阿维菌素未产生明显的抗药性;烟粉虱田间种群对烟碱类杀虫剂的抗性发展极为迅速,其中漳州种群对吡虫啉、噻虫嗪已由2005年的中等水平抗性（抗性倍数分别为23和25倍）发展为2009年的高水平抗性（抗性倍数分别为103和228倍）,其他地区种群对吡虫啉、噻虫嗪也由2005年的低水平抗性（抗性倍数分别为1.5～3.3倍和1.7～5.5倍）发展为2009年的中等水平抗性（抗性倍数分别为23～33倍和29～49倍）.采用mtDNA CO I分子标记技术对福建省不同地区和寄主植物上采集的8个烟粉虱种群的生物型进行鉴定发现,采自漳州变叶木上的烟粉虱种群为本地土著种(未知生物型),其他7个采自大田蔬菜作物的烟粉虱种群均为B型.寄主植物和温度对烟粉虱种群的药剂敏感性影响有限,吡虫啉等烟碱类杀虫剂高强度使用是B型烟粉虱田间种群对该类型药剂迅速形成抗性的重要原因.     

蜡蚧轮枝菌毒素对温室中烟粉虱种群控制作用的评价

在室内测定蜡蚧轮枝菌毒素对烟粉虱种群干扰作用基础上,通过分别组建蜡蚧轮枝菌毒素和化学杀虫剂作用下的烟粉虱第5、6代自然种群生命表,采用种群趋势指数（I）和干扰作用控制指数（IIPC）分析法,比较蜡蚧轮枝菌毒素和化学杀虫剂对茄子上烟粉虱的防治效果,评价蜡蚧轮枝菌毒素对烟粉虱的防治作用。结果表明,毒素对烟粉虱室内种群的干扰作用主要表现在对成虫的忌避作用和对若虫的毒杀作用;温室大棚中施用400mg／L的蜡蚧轮枝菌毒素,对烟粉虱第5代和第6代的控制指数IIPC分别为0．064和0．023,连续施用毒素后,第6代种群趋势指数Ⅰ为0．68,烟粉虱种群基本得到控制;温室大棚中施用重量10％吡虫啉可湿性粉剂（稀释1000倍）,对烟粉虱第5代和第6代的控制指数IIPC分别为0．44和1．01,化防区第5代的I为12．95,第6代的I为30．23,分别为对照区的0．44倍和1．01倍,连续使用化学杀虫剂,容易造成烟粉虱再猖獗。重要因子分析揭示毒索比化学杀虫剂更利于温室烟粉虱种群控制。     

烟粉虱B型与Q型种群遗传多样性的AFLP分析

烟粉虱Bemisia tabaci(Gennadius)是由多种生物型或物种组成的复合种。Q型烟粉虱在我国部分地区正在取代B型烟粉虱成为优势生物型。烟粉虱Q型与B型遗传多样性比较研究为解析这2种生物型入侵的遗传学基础具有重要的意义。本文利用AFLP技术研究了Q型(包括Q1型、Q2型)、B型烟粉虱种群的遗传多样性。结果表明:Q型烟粉虱遗传多样性高于B型烟粉虱;Q1型烟粉虱各项遗传多样性指数均接近于Q2型。最后探讨了AFLP与SSR分子标记的各自特点。     

微卫星位点在Q型烟粉虱入侵种群中的多态性分析

近年来烟粉虱Bemisia tabaci(Gennadius)Q型入侵我国并在部分地区取代B型成为了烟粉虱优势生物型。外来物种的入侵过程及农药使用等因素可影响种群的遗传多样性水平及其遗传结构。本研究分析了4个微卫星位点在Q型烟粉虱入侵种群的多态信息含量(PIC),并在此基础上进行了遗传多样性分析。结果表明这些微卫星位点在Q型烟粉虱入侵种群中具有中度或高度多态性,说明微卫星位点能有效分析Q型入侵种群的遗传多态性。本研究为利用微卫星标记研究Q型入侵种群的遗传结构奠定了基础。     

B型烟粉虱与浙江非B型烟粉虱的竞争

为了了解近年来入侵中国的B型烟粉虱(Bemisiatabaci)取代本地非B型烟粉虱的潜能,在室内将一个B型与一个浙江非B型烟粉虱种群混合饲养在不同寄主植物上,跟踪观察混合种群中两个生物型个体数量相对比例的变化。结果表明,当两种生物型在棉花(Gossypiumhirsutum)上以相同初始数量共存竞争时,经过6代,非B型完全被B型替代;而在西葫芦(Cucurbitapepo)上以相同初始数量共存竞争时,只经过2代,非B型即完全被B型替代。在棉花上,即使以非B型占87%、B型占13%开始共存竞争,经过225d后,非B型也完全被B型替代。这说明B型烟粉虱具有在短期内竞争取代浙江非B型烟粉虱的能力。经分析,B型除了寄主范围比非B型的宽这一点对其竞争有利外,较强的内在竞争潜能也是其能成功入侵并替代本地非B型烟粉虱的一个重要原因。     

Biotypes B and Q of Bemisia tabaci and their relevance to neonicotinoid and pyriproxyfen resistance

Resistance monitoring for Bemisia tabaci field populations to the juvenile hormone mimic, pyriproxyfen, was conducted from 1996 to 2003 in commercial cotton fields in two areas of Israel: the Ayalon Valley (central Israel) and the Carmel Coast (northwestern Israel). Although the use of pyriproxyfen ceased in these areas in 1996-1997 (because of the resistance), resistance levels to pyriproxyfen declined to some extent in the fields but remained quite stable, and the susceptibility has not been totally restored. Two strains of B. tabaci collected from the Ayalon Valley in the late 1999 and 2002 cotton seasons (AV99L, AV02L) were assayed for their susceptibility to pyriproxyfen at F1, and subsequently a line of each strain was kept under controlled conditions without exposure to insecticides. After maintenance of more than 20 generations under laboratory conditions, the resistance to pyriproxyfen in the untreated strains substantially declined. This decline was concurrent with a replacement of Q biotype by B-type under non-insecticidal regimes; apparently B biotype was more competitive than the pyriproxyfen-resistant Q-type. Selection under controlled conditions with neonicotinoids on these B. tabaci strains resulted in continued pyriproxyfen resistance, predominantly of Q biotype. Based on our data, applications of either pyriproxyfen or neonicotinoids may select for biotype Q, which would survive to a greater degree where these insecticides are applied.     

Dynamics of resistance to the neonicotinoids acetamiprid and thiamethoxam in Bemisia tabaci (Homoptera: Aleyrodidae)

The dynamics of resistance in the sweetpotato whitefly, Bemisia tabaci (Gennadius), to the neonicotinoids acetamiprid and thiamethoxam was studied extensively in cotton fields in Israel during the cotton-growing seasons 1999-2003. Whitefly strains were collected in early and late seasons mainly in three locations in northern, central, and southern Israel. The whiteflies were assayed under laboratory conditions for susceptibility to neonicotinoids, as part of the Israeli cotton insecticide resistance management strategy. Selections to both acetamiprid and thiamethoxam and cross-resistance between them also were conducted in the laboratory. Although no appreciable resistance to acetamiprid was observed up to 2001, a slight increase of approximately five-fold resistance was detected during 2002 and 2003. However, from 2001 to 2003 thiamethoxam resistance increased >100-fold in the Ayalon Valley and Carmel Coast cotton fields. In cross-resistance assays with both neonicotinoids, the strain that had been selected with thiamethoxam for 12 generations demonstrated almost no cross-resistance to acetamiprid, whereas the acetamiprid-selected strain exhibited high cross-resistance of >500-fold to thiamethoxam.     

Resistance of insect pests to neonicotinoid insecticides: current status and future prospects

The first neonicotinoid insecticide introduced to the market was imidacloprid in 1991 followed by several others belonging to the same chemical class and with the same mode of action. The development of neonicotinoid insecticides has provided growers with invaluable new tools for managing some of the world's most destructive crop pests, primarily those of the order Hemiptera (aphids, whiteflies, and planthoppers) and Coleoptera (beetles), including species with a long history of resistance to earlier-used products. To date, neonicotinoids have proved relatively resilient to the development of resistance, especially when considering aphids such as Myzus persicae and Phorodon humuli. Although the susceptibility of M. persicae may vary up to 20-fold between populations, this does not appear to compromise the field performance of neonicotinoids. Stronger resistance has been confirmed in some populations of the whitefly, Bemisia tabaci, and the Colorado potato beetle, Leptinotarsa decemlineata. Resistance in B- and Q-type B. tabaci appears to be linked to enhanced oxidative detoxification of neonicotinoids due to overexpression of monooxygenases. No evidence for target-site resistance has been found in whiteflies, whereas the possibility of target-site resistance in L. decemlineata is being investigated further. Strategies to combat neonicotinoid resistance must take account of the cross-resistance characteristics of these mechanisms, the ecology of target pests on different host plants, and the implications of increasing diversification of the neonicotinoid market due to a continuing introduction of new molecules.     

镶嵌式防治对抗性演化影响的论证

为了评估镶嵌式防冶对抗性演化的影响,我们以淡色库蚊(Culex pipiens pallens)作为模型昆虫,用抗马拉硫磷(单因子)品系(RM)和抗氰戊菊酯(多因子)品系(RF)构成抗性个体频率均为0.1的一个“什成群体”(M品系),然后在F₁代分成三个亚品系。Rmf亚品系用马拉硫磷和氰戊菊酯作镶嵌式模拟处理,并与单用马拉硫磷(Rm)和氰戊菊酯(Rf)逐代处理的亚品系作比较。处理10代后,Rm和Rmf对马拉硫磷的抗性分别是M晶系的266.4和6.1倍,而Rf和Rmf在处理5代后,对氰戊菊酯;山抗性均已达243.3倍。漠拟结果表明：(1)以杀虫饥理不同的杀虫剂进行镶嵌式防冶,在一定的条件下能延缓抗性的发展;(2)对单因子遗传,内马拉硫磷特别有效,而对多因子遗传的氰戊菊酯效果不佳。此外,还讨论了抗性治理的策略。     

Development of neonicotinoid resistance in the cotton aphid Aphis gossypii (Hemiptera: Aphididae) in Japan

A neonicotinoid-resistant Aphis gossypii Glover population was first detected from cucumber and sweet pepper crops in Miyazaki Prefecture, Japan, in April 2012. In this study, we determined the insecticide susceptibility of five field-collected populations of A. gossypii to seven neonicotinoids. Insecticide susceptibility was determined by performing bioassays and using the seedling-treatment method. Insecticide susceptibility of a susceptible population of A. gossypii, originally collected from a cucumber field in 2008, was also determined. High mortality (96.4–100 %) was observed in the susceptible population for the seven insecticides. In contrast, the mortality rate of the five field-collected populations collected in 2012 was low for five of the seven neonicotinoid insecticides tested: imidacloprid (26.7–65.5 %), dinotefuran (0–27.3 %), clothianidin (20.0–35.7 %), thiamethoxam (7.1–42.3 %), and nitenpyram (6.7–32.1 %). Mortality was 86.2–100 % for acetamiprid and 90.2–100 % for thiacloprid. The resistance ratio in comparison with the susceptible population was the highest for clothianidin (687-fold), lowest for thiacloprid (17-fold), and ranged from 43- to 253-fold for the other five neonicotinoids. Thus, the frequency of insecticide use needs to be reduced and other insecticides need to be employed in order to prevent the development of resistance to these two insecticides in the future.     

Stability of spinosad resistance in Frankliniella occidentalis (Pergande) under laboratory conditions

The stability of spinosad resistance in western flower thrips (WFT), Frankliniella occidentalis (Pergande), populations with differing initial frequencies of resistance was studied in laboratory conditions. The stability of resistance was assessed in bimonthly residual bioassays in five populations with initial frequencies of 100, 75, 50, 25 and 0% of resistant individuals. There were no consistent changes in susceptibility of the susceptible strain after eight months without insecticide pressure. In the resistant strain, very highly resistant to spinosad (RF50>23,000-fold), resistance was maintained up to eight months without further exposure to spinosad. In the absence of any immigration of susceptible genes into the population, resistance was stable. In the case of the population with different initial frequency of resistant thrips, spinosad resistance declined significantly two months later in the absence of selection pressure. With successive generations, these strains did not change significantly in sensitivity. Spinosad resistance in F. occidentalis declined significantly in the absence of selection pressure and the presence of susceptible WFT. These results suggest that spinosad resistance probably is unstable under field conditions, primarily due to the immigration of susceptible WFT. Factors influencing stability or reversion of spinosad resistance are discussed.     

Changes in emergence phenology,fatty acid composition,and xenobiotic-metabolizing enzyme expression is associated with increased insecticide resistance in the Colorado potato beetle

The Colorado potato beetle (Leptinotarsa decemlineata) is a major agricultural pest of solanaceous crops. An effective management strategy employed by agricultural producers to control this pest species is the use of systemic insecticides. Recent emphasis has been placed on the use of neonicotinoid insecticides. Despite efforts to curb resistance development through integrated pest management approaches, resistance to neonicotinoids in L. decemlineata populations continues to increase. One contributing factor may be alterations in insect fatty acids, which have multiple metabolic functions and are associated with the synthesis of xenobiotic-metabolizing enzymes to mitigate the effects of insecticide exposure. In this study, we analyzed the fatty acid composition of L. decemlineata populations collected from an organic production field and from a commercially managed field to determine if fatty acid composition varied between the two populations. We demonstrate that a population of L. decemlineata that has a history of systemic neonicotinoid exposure (commercially managed) has a different lipid composition and differential expression of known metabolic detoxification mechanisms relative to a population that has not been exposed to neonicotinoids (organically managed). The fatty acid data indicated an upregulation of Δ⁶ desaturase in the commercially managed L. decemlineata population and suggest a role for eicosanoids and associated metabolic enzymes as potential modulators of insecticide resistance. We further observed a pattern of delayed emergence within the commercially managed population compared with the organically managed population. Variations in emergence timing together with specific fatty acid regulation may significantly influence the capacity of L. decemlineata to develop insecticide resistance.     

Inheritance of lambda-cyhalothrin resistance in the predator lady beetle Eriopis connexa (Germar) (Coleoptera: Coccinellidae)

Lambda-cyhalothrin is a pyrethroid insecticide widely used to control pests in various crop ecosystems, in which the lady beetle occurs naturally. Therefore, lady beetle populations are exposed to lambda-cyhalothrin sprays that may foster tolerance to this insecticide. This study was conducted to confirm the occurrence of resistance in the lady beetle Eriopis connexa (Germar) (Coleoptera: Coccinellidae) to lambda-cyhalothrin and to characterize the inheritance of resistance after eight progressive selection with insecticide dosages based on the LD₅₀ determined for the F1 generation. Dose–mortality curves were determined for parental populations, F1 hybrids and backcross progenies. Parameters regarding the heritability of resistance to lambda-cyhalothrin in E. connexa allow estimating 10-fold increase in the initial LD₅₀ after 54.5 generations of successive selections. The resistance of E. connexa to lambda-cyhalothrin was characterized as autosomally inherited and incompletely dominant, and influenced by a major gene with possible influence of secondary genes. Additionally, the resistance in E. connexa varies from functionally dominant to functionally recessive depending on the dose used. These findings indicate that insecticide resistance in E. connexa can be selected in the field as determined for field-collected individuals, and subsequently enhanced under laboratory conditions. Its characterization presented here is an important step toward linking biological and chemical control within pest management regarding the lady beetle and lambda-cyhalothrin targeting different pest groups.     

韭菜迟眼蕈蚊对新烟碱类杀虫剂的抗药性及抗性机制初探

【目的】为明确韭菜迟眼蕈蚊Bradysia odoriphaga对新烟碱类杀虫剂的抗性水平及其抗性机制。【方法】通过测定不同地区韭菜迟眼蕈蚊对3种新烟碱类杀虫剂吡虫啉、噻虫嗪和噻虫胺的敏感度,及通过增效剂实验和酶活性测定,初步探索抗性产生机制,为韭菜迟眼蕈蚊抗性治理提供依据。【结果与结论】4个不同的韭菜迟眼蕈蚊田间种群对3种新烟碱类杀虫剂均产生了不同水平的抗性。其中,唐山种群对3种新烟碱杀虫剂均产生了较高的抗性。研究发现,唐山种群的7-乙氧基香豆素-O-脱乙基酶(ECOD)比活力为(3.89±0.31)pmol/(mg·pro·min),显著高于敏感品系。增效剂PBO对唐山种群的吡虫啉毒力的增效比为2.64,高于对敏感品系的增效比1.08。因此,P450s酶活性的升高与韭菜迟眼蕈蚊对吡虫啉的抗性有关。