mtCOI PCR-RFLP技术鉴别中国入侵型和土著型烟粉虱种群的有效性分析

烟粉虱Bemisia tabaci (Gennadius)是世界范围内最重要的入侵生物之一,准确地鉴别烟粉虱入侵生物型和土著生物型具有十分重要的现实意义。mtCOI PCR-RFLP技术具有快速、高效的特点,是当前应用最广泛的烟粉虱生物型鉴定技术,但是,不同限制性内切酶为基础的mtCOI PCR-RFLP技术在鉴定我国烟粉虱种群中的有效性仍不明了。本文比较了已报导的5种mtCOI PCR-RFLP技术在鉴别中国烟粉虱种群入侵生物型和土著生物型(B型、Q型,ZHJ1型、ZHJ2型、ZHJ3型)的有效性。结果表明,内切酶AluI不能区分B型和ZHJ2型烟粉虱;内切酶TaqI不能准确区分ZHJ3型和Q型烟粉虱个体;而内切酶VspI不仅不能准确区分ZHJ1型和Q型烟粉虱个体,也不能准确区分B型和ZHJ2型烟粉虱;内切酶MseI和Tru9I则不能有效鉴别上述5种烟粉虱生物型,因此不适宜推广使用。     

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。     

B型烟粉虱的入侵机理与控制基础——国家重点基础研究发展计划“农林危险生物入侵机理与控制基础研究”进展

针对B型烟粉虱入侵后的暴发性及生态系统的可侵入性,开展了B型烟粉虱的入侵机制、成灾机理和控制基础研究.结果显示,目前我国至少存在6种生物型（B,Q,ZHJ-1,ZHJ-2,ZHJ-3,FJ-1）,其中B型烟粉虱在我国大部分地区、Q型烟粉虱在我国长江流域已成为优势种群,给蔬菜、花卉和棉花生产造成严重危害.寄主植物、地理环境及杀虫剂可诱导B型烟粉虱产生遗传分化,使其种群遗传结构发生快速演变;黄瓜和南瓜可诱导羧酸酯酶（CarE）和谷胱甘肽S-移酶（GSTs）活性升高,增强B型烟粉虱抗药性;植物次生物可诱导B型烟粉虱解毒酶活性升高.B型烟粉虱所具有的较强的高温胁迫适应能力和热激蛋白基因的表达与响应密切相关.B型烟粉虱可取代土著烟粉虱,B型烟粉虱与土著烟粉虱间的非对称交配干扰及其与植物双生病毒间的互利共生加剧了B型烟粉虱的入侵;特定条件下B型烟粉虱可取代温室粉虱,较强的逆境适应能力、较快的种群增长力可能是B型烟粉虱取代温室粉虱的主要机制;较强的对寄主转换的适应能力与B型烟粉虱寄主谱扩张有关.土著天敌对B型烟粉虱具有较强的控制潜能;B型烟粉虱3,4龄若虫及蜜露可诱导丽蚜小蜂产生强烈的搜索行为,若虫利它素在丽蚜小蜂寄主搜索和定位中具有重要作用.研究结果为阐明B型烟粉虱入侵种群在我国的遗传分化与快速演变机制和分子生态适应机制、探明种群形成与扩张过程中生态对策的调整及其效应提供了理论依据,为B型烟粉虱的生物生态控制途径和持续治理策略提供了保障。     

广东首次发现Q型烟粉虱种群分布

烟粉虱Bemisia tabaci(Gennadius)是全世界分布的刺吸式传毒昆虫,其中Q型烟粉虱自2003年首次在昆明一品红上发现以来,该种群已在全国10多个省份广泛传播,并有逐步取代B型烟粉虱的趋势。然而,作为较早对外开放的沿海省份,广东省一直未有Q型烟粉虱在田间分布危害的报道。本文利用常规PCR、特异B/Q型烟粉虱引物扩增及DNA测序技术,对近期在广州长湴公园扶桑寄主上采集的烟粉虱种群进行了分子鉴定,并对该种群与国内外其它Q型种群的系统发育关系进行了研究。结果表明,在广州长湴公园扶桑寄主上采集的烟粉虱种群为Q型烟粉虱,在660 bp的mtCOI序列中,该Q型烟粉虱与来自江苏、云南及浙江的Q型烟粉虱种群分别有1²个碱基差异;系统进化关系表明该Q型烟粉虱与江苏、云南的Q型烟粉虱最近,其种群的最初起源应为地中海西岸地区。相比B型烟粉虱,Q烟粉虱被认为具有更强的抗药性和传播植物病毒的能力,Q型烟粉虱在广东的首次发现,表明该烟粉虱种群已在广东野外定殖,其扩散动态、寄主范围及危害程度亟待实时监测。     

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。     

世界性重要害虫B型烟粉虱的入侵机制

B型烟粉虱是近二十年来入侵世界各国并暴发成灾的一种世界性重要害虫。近年来, B型烟粉虱的入侵机制研究表明,其成功定殖、扩散、暴发与其对入侵地区的土著烟粉虱种群和其它昆虫种群的竞争取代有关;竞争取代包括生态位竞争,生殖干涉,其危害寄主对其它昆虫的影响等方面。B型烟粉虱抗药性的形成也是这种生物成功入侵的重要影响因素。同时,生态环境（包括寄主植物、气候因子、越冬场所等）的影响也不容忽视。最后本文还探讨了B型烟粉虱入侵机制研究动态及其意义。     

微卫星位点BEM06与BEM23鉴别烟粉虱B型与Q型的有效性

烟粉虱Bemisia tabaci (Gennadius) B型与Q型是烟粉虱复合种中入侵性较强、分布较广的2种生物型, 当前在许多地区混合发生。这2种生物型的快速鉴别对其种群动态调查及入侵生态学研究具有重要价值。为了验证微卫星位点BEM06与BEM23鉴别B型与Q型烟粉虱的有效性, 本研究分析了这2个微卫星位点的等位基因在国内外17个B型、4个Q型、3个非B/Q型烟粉虱种群的分布特点。结果表明: 这2个微卫星位点的联合使用可鉴别B型与Q型烟粉虱, 但是无法有效地将B型、Q型与一些非B/Q型烟粉虱某些个体区分开来。结果提示: 利用微卫星位点BEM06与BEM23鉴别B型与Q型烟粉虱具有一定的局限性, 尤其是田间烟粉虱存在其他生物型时需要慎重使用这种鉴别方法。     

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。     

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。     

B型烟粉虱不同寄主种群伪蛹形态的可塑性比较

烟粉虱Bemisia tabaci(Gennadius)是一种包含多个生物型的复合种,其中B型烟粉虱是一个入侵能力较强的生物型。对在不同寄主植物上B型烟粉虱入侵种群伪蛹形态系统量化研究结果表明:B型烟粉虱伪蛹蛹壳规则比例、蛹壳背部大刚毛数量、蛹壳整体的长度和宽度、前缘蜡饰的宽度、后缘蜡饰的宽度、皿形孔盖瓣的长度和宽度以及舌状突的长度等许多性状在不同寄主植物上具有不同程度的形态可塑性,而且这种可塑性与寄主植物叶背特征具有密切的相关性。最后,探讨B型烟粉虱伪蛹形态可塑性在该害虫成功入侵过程中的作用以及研究意义。     

江苏地区烟粉虱生物型演替研究初报

本文连续5年系统监测了江苏省13个地级市烟粉虱Bemisia tabaci(Gennadius)生物型的发生分布状况,探讨了它们的迁移扩散和演替规律。利用RAPD分子标记和mtDNA COI基因序列进行烟粉虱生物型鉴定,结果表明:江苏地区发生的烟粉虱生物型为B型和Q型。自2005年到2009年,B型烟粉虱在苏南和苏北地区的发生分布频率逐年下降,在苏北地区的发生分布频率由52.58%下降为22.22%,在苏南由56.52%下降为9.37%;而Q型烟粉虱在苏南和苏北的发生分布频率逐年升高,在苏北由47.42%上升至77.78%,在苏南由43.48%上升为90.63%。B型烟粉虱的发生分布范围由全省逐渐向苏北地区缩小,而Q型烟粉虱的发生分布范围逐渐扩大遍及全省;此外,Q型烟粉虱在江苏是由苏中和苏南地区向苏北地区扩散,并逐步取代B型烟粉虱成为江苏地区农作物的主要害虫。     

Biotype-dependent secondary symbiont communities in sympatric populations of Bemisia tabaci

The sweet potato whitefly, Bemisia tabaci, harbors Portiera aleyrodidarum, an obligatory symbiotic bacterium, as well as several secondary symbionts including Rickettsia, Hamiltonella, Wolbachia, Arsenophonus, Cardinium and Fritschea, the function of which is unknown. Bemisia tabaci is a species complex composed of numerous biotypes, which may differ from each other both genetically and biologically. Only the B and Q biotypes have been reported from Israel. Secondary symbiont infection frequencies of Israeli laboratory and field populations of B. tabaci from various host plants were determined by PCR, in order to test for correlation between bacterial composition to biotype and host plant. Hamiltonella was detected only in populations of the B biotype, while Wolbachia and Arsenophonus were found only in the Q biotype (33% and 87% infection, respectively). Rickettsia was abundant in both biotypes. Cardinium and Fritschea were not found in any of the populations. No differences in secondary symbionts were found among host plants within the B biotype; but within the Q biotype, all whiteflies collected from sage harboured both Rickettsia and Arsenophonus, an infection frequency which was significantly higher than those found in association with all other host plants. The association found between whitefly biotypes and secondary symbionts suggests a possible contribution of these bacteria to host characteristics such as insecticide resistance, host range, virus transmission and speciation.     

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。     

MtDNA variability in whitefly (Bemisia tabaci) populations in Brazil

Bemisia tabaci (Hemiptera: Aleyrodidae) consists of a complex of morphologically indistinct biotypes that vary mainly in their capacity to transmit plant viruses and to induce physiological disorders in plants of economic importance. The adaptability of B. tabaci to many regions of the world has fostered the appearance of various biotypes and has resulted in a broad spectrum of host plants. Our goal was to identify which biotypes were present in four B. tabaci populations in Brazil. We quantified genetic variability between and within populations. Three individuals were collected from three host plant species: two populations on soybean (Campinas and Rondonópolis), one on pumpkin (Barreiras) and one on tomato (Cruz das Almas) in three States of Brazil (S?o Paulo, Mato Grosso, and Bahia). We chose one sequence of the B biotype, obtained from GenBank; the Campinas population, which had been previously characterized as biotype B, was used as a control for this biotype. We also included one sequence of the Q biotype, obtained from GenBank, as an outgroup. The COI region of the mtDNA gene was partially amplified with the CI-J-2195 and L2-N-3014 pair of primers, and the reaction products were sequenced. Based on distance-based algorithm analyses, we found that all haplotypes belong to biotype B, which was confirmed by the haplotype network. Genetic structure analyses showed that the host plant species does not influence population structuring of this pest; only the geographic location mattered.     

Influence of plant combinations on population characteristics of Bemisia tabaci biotypes B and Q

In recent years, the dominant biotype of Bemisia tabaci (Gennadius) has shifted from biotype B to Q in many locations of China. Our field monitoring showed that B. tabaci biotype Q could be found on pepper Capsicum frutescens L. while biotype B could not be found on the plant. To study the role of the host plant in the displacement of biotype B by Q, the adult mortality, female fecundity, and adult emergence of both biotypes B and Q on different host combinations were studied using a model system involving pepper and eggplant. The results showed that pepper is not a preferred host for either biotype B or Q adults in comparison with eggplant. The negative impact of pepper to biotype B is, however, much greater than that to biotype Q. Because both the survival rates of the adult whitefly and the average number of emerged adults per female of biotype Q on pepper are higher than that of biotype B, biotype Q showed higher fitness in comparison with biotype B. Our results suggest that the existence and differential impact of a nonpreferred host might affect the population fitness and therefore could play a role in the displacement of biotype B in some regions.     

不同作物田烟粉虱发生的时空动态

2008～2009年连续2年系统调查了番茄、茄子、棉花、大豆、玉米等寄主植物上烟粉虱种群发生的时间与空间动态。结果表明：不同寄主植物上的烟粉虱成虫及其伪蛹数量有显著性差异,其密度大小依次为：茄子>棉花>番茄>大豆>玉米。其中,在玉米上除了发现极少量的成虫逗留外,没有发现烟粉虱的卵及若虫。在发生的时间序列上,烟粉虱成虫及伪蛹的数量呈现为先逐渐上升后又下降的变化过程,发生高峰期集中在8月5日到8月31日,9月初以后烟粉虱数量慢慢减少。在空间分布上,表现为烟粉虱成虫喜食寄主的上部叶片。统计分析显示,寄主对烟粉虱成虫和伪蛹的数量的影响极显著,而年份对其数量的影响没有显著差异。由此得出的烟粉虱发生和达到高峰的时间,可为烟粉虱预测预报和区域性综合治理提供重要理论依据。     

Analysis of genetic diversity among different geographical populations and determination of biotypes of Bemisia tabaci in China

Abstract:  Analysis of the genetic diversity among 27 different geographical populations of Bemisia tabaci and determination of biotypes of B. tabaci in China based on amplified fragment-length polymorphism (AFLP) and the mitochondrial cytochrome oxidase I (mtDNA COI) gene sequences were conducted. In AFLP assay, the use of five primer combinations selected from 64 primer combinations allowed the identification of 229 polymorphic bands (97.03%) from 60 to 500 bp, suggesting abundant genetic diversity among different geographical populations of B. tabaci. To further identify biotypes of B. tabaci in China, the mtDNA COI gene sequences of nine representative populations from China, Israel and Spain were obtained. Molecular phylogenetic tree based on AFLP and mtDNA COI gene analyses revealed the presence, in China, of at least four different genetic groups of B. tabaci. B biotype, Q biotype and two non-B/Q biotype. B biotype was distributed nationwide. Q biotype was present only in the local region of China including the YunNan province and BeiJing city. This was also the first report about the invasion of Q biotype into China. Of the other two non-B/Q biotype groups, one was found in ShanDong and HeBei provinces, and another in ZheJiang province. The non-B/Q biotype ZheJiang population showed very high similarity with another Asian population India-IW ( AF110704 ) in mtDNA COI sequences and was possibly a Chinese indigenous population. The close monitoring of the Q biotype in locales of China where commercial plants were exported or imported, is now essential to avoid the further accidental distribution of the Q biotype.     

Invasive mechanism and management strategy of Bemisia tabaci (Gennadius) biotype B: Progress report of 973 Program on invasive alien species in China

Bemisia tabaci (Gennadius) biotype B, called a “superbug”, is one of the most harmful biotypes of this species complex worldwide. In this report, the invasive mechanism and management of B. tabaci bio-type B, based on our 5-year studies, are presented. Six B. tabaci biotypes, B, Q, ZHJ1, ZHJ2, ZHJ3 and FJ1, have been identified in China. Biotype B dominates the other biotypes in many regions of the country. Genetic diversity in biotype B might be induced by host plant, geographical conditions, and/or insecticidal application. The activities of CarE (carboxylesterase) and GSTs (glutathione-S-transferase) in biotype B reared on cucumber and squash were greater than on other host plants, which might have increased its resistance to insecticides. The higher activities of detoxification enzymes in biotype B might be induced by the secondary metabolites in host plants. Higher adaptive ability of biotype B adults to adverse conditions might be linked to the expression of heat shock protein genes. The in-digenous B. tabaci biotypes were displaced by the biotype B within 225 d. The asymmetric mating in-teractions and mutualism between biotype B and begomoviruses via its host plants speed up wide-spread invasion and displacement of other biotypes. B. tabaci biotype B displaced Trialeurodes vapo-rariorum (Westwood) after 4－7 generations under glasshouse conditions. Greater adaptive ability of the biotype B to adverse conditions and its rapid population increase might be the reasons of its suc-cessful displacement of T. vaporariorum. Greater ability of the biotype B to switch to different host plants may enrich its host plants, which might enable it to better compete with T. vaporariorum. Native predatory natural enemies possess greater ability to suppress B. tabaci under field conditions. The kairomones in the 3rd and 4th instars of biotype B may provide an important stimulus in host searching and location by its parasitoids. The present results provide useful information in explaining the mechanisms of genetic diversity, evolution and molecular eco-adaptation of biotype B. Furthermore, it provides a base for sustainable management of B. tabaci using biological and ecological measures.