烟粉虱抗、感杀虫剂品系内生菌群落的差异分析

本研究对B型和Q型烟粉虱的吡虫啉或吡丙醚的抗、感品系进行16SrRNA细菌多样性测序分析,以明确内共生菌的种类和数量与烟粉虱生物型和抗性的关系。结果表明Candidatus Nitrososphaera、Bacillus、Sphingobium只在Q型烟粉虱中存在,而Candidatus Hamiltonella、Lactobacillus、Helicobacter等只在B型烟粉虱中存在,说明这些内共生菌可能与烟粉虱B、Q两种生物型的分化有关;内共生菌Rickettsia、Candidatus Portiera、Candidatus Hamiltonella、S24-7、Prevotella等在烟粉虱的新烟碱类杀虫剂吡虫啉抗性品系中明显多于敏感品系,其可能与烟粉虱对吡虫啉的抗性有关;在对保幼激素类似物杀虫剂吡丙醚的抗性品系中,Candidatus Hamiltonella-OTU7、Candidatus Portiera-OTU5、Candidatus Portiera-OTU190、Rickettsia-OTU4、Rickettsia-OTU2的数量明显多于敏感品系,说明这些内共生菌可能与烟粉虱对该类药物的抗性有关。     

天津市烟粉虱隐种鉴定及其携带TYLCV、内共生菌情况和抗药性监测

为明确天津市烟粉虱Bemisia tabaci隐种的类别及其寄主适应性、传毒能力、携带内共生菌情况和抗药性,采用mt COI酶切法对从武清、西青、蓟州和宁河4个区的番茄、黄瓜及辣椒3种寄主上采集的12个烟粉虱种群进行隐种鉴定,采用PCR检测其携带番茄黄化曲叶病毒（tomato yellow leaf curl virus,TYLCV）和内共生菌情况,并采用浸叶法测定其对4种常用药剂的抗性。结果表明,采集的烟粉虱种群以MED隐种为主,占所有检测个体的93.33%,有3个种群为MED和MEAM1隐种混合发生。所有检测个体中有36.25%的个体携带TYLCV,在6个种群中检测到TYLCV,其中5个种群有超过50%的个体携带TYLCV。在12个种群中共检测到Hamiltonella、立克次氏体Rickettsia、Cardinium和杀雄菌属Arsenophnus共4种内共生菌,携带个体比例分别为90.63%、48.96%、43.75%和8.33%,进一步对内共生菌协同感染情况进行分析,发现有HARC、HRC、HAC、HR、HC和AC共6个协同感染型,感染率分别为4.17%、28.13%、3....     

烟粉虱基因组和转录组研究及展望

烟粉虱Bemisia tabaci(Gennadius)是世界性的重要害虫,为物种复合体,其中少数隐种近30年来入侵世界各国,并暴发成灾。在我国,最近几年MED隐种已经逐渐替代了之前入侵的MEAM1隐种,成为我国农田生态系统中的主要致害隐种。基因组和转录组研究的不断深入极大地推动了烟粉虱入侵机制、隐种替代机制、种系发生、不同隐种或地理种群鉴定、抗药性机制、烟粉虱与共生菌互作等领域的研究。本文简要综述了烟粉虱基因组和转录组测序的研究进展,介绍了转录组数据在种的分化和鉴定、共生菌的互作及抗药性研究领域的应用。     

肿瘤坏死因子受体相关因子基因TRAF4促进烟粉虱与共生菌Portiera的共生关系

为明确肿瘤坏死因子受体相关因子（tumor necrosis factor receptor-associated factor,TRAF）基因TRAF4在烟粉虱Bemisia tabaci隐种MEAM1与含菌细胞共生菌互作中的作用,对TRAF4蛋白进行保守结构域及系统发育分析;通过实时荧光定量PCR（quantitative real-time PCR,qPCR）方法检测TRAF4在含菌细胞和整虫体内的表达量;利用RNA干扰（RNA interference,RNAi）技术抑制烟粉虱TRAF4基因表达,然后通过qPCR技术检测含菌细胞共生菌滴度、统计烟粉虱产卵量以及采用免疫荧光技术检测烟粉虱含菌细胞的自噬水平。结果表明,TRAF4蛋白含有Zinc finger和TRAF结构域,与黑腹果蝇 Drosophila melanogaster 及埃及伊蚊 Aedes aegypti的 TRAF4 蛋白聚为一支。TRAF4在含菌细胞中显著高表达。注射dsTRAF4后,TRAF4基因表达量显著下调,Portiera滴度及烟粉虱产卵量均显著下降,含菌细胞自噬水平显著升高。表明烟粉虱TRAF4不仅影响烟粉虱的产卵量,还能通过抑制含菌细胞的自噬水平维持Portiera在烟粉虱体内的滴度,对烟粉虱与Portiera共生关系的维持起到正向调控作用。     

传播木尔坦棉花曲叶病毒的烟粉虱隐种鉴定

为明确广东地区传播木尔坦棉花曲叶病毒(Cotton leaf curl Multan virus,CLCu Mu V)的烟粉虱隐种,采用分子鉴定、烟粉虱传毒试验与分子检测的方法,对传播CLCu Mu V的烟粉虱隐种进行了鉴定。结果表明:在广东棉花、红麻和黄秋葵曲叶病发生的田间,2014年采集的30头烟粉虱mt COI与Asia II 7隐种序列同源性为98.3%~99.6%,2015年采集的10头烟粉虱中,8头烟粉虱mt COI与MEAM1隐种序列同源性为99.2%~99.5%,2头烟粉虱mt COI与Asia II 7隐种序列同源性为99.8%~99.9%,说明烟粉虱种群包括入侵隐种MEAM1和土著隐种Asia II 7。在利用烟粉虱人工传毒试验中,MEAM1、Asia II 7和Asia II 1这3个隐种均可在棉花曲叶病株上饲毒获得CLCu Mu V及β卫星分子;除MEAM1隐种外,Asia II 7、Asia II 1隐种可传播CLCu Mu V,侵染红麻及黄秋葵植株引起曲叶病,前者传毒效率分别为50%和100%;而后者为33%和100%。以上3个烟粉虱隐种传毒接种棉花未见成功。     

节肢动物共生细菌Hamiltonella的分布、传播、功能及基因组研究

Hamiltonella是节肢动物体内的一种次生共生细菌,目前主要在蚜虫和粉虱体内发现,其有两种传播方式,即随母系生殖垂直传播和在寄主个体间水平传播。在自然界中,Hamiltonella在蚜虫中的侵染率一直维持在中等水平,而在烟粉虱物种复合体中仅感染入侵隐种B和Q。Hamiltonella在寄主体内的功能研究还很匮乏,目前已知的功能主要是与寄主防御反应相关,蚜虫受Hamiltonella侵染后,其抵抗寄生蜂寄生的能力显著增加。直接的生物学性状观察没有发现Hamiltonella对豌豆蚜有不利影响,相反,感染Hamiltonella的豌豆蚜生活力更强。但Hamiltonella在黑豆蚜中表现出对寄主不利的现象。除了影响寄主抗逆能力,Hamiltonella也可以对植物病毒在烟粉虱体内的循环提供保护,从而有利于植物病毒的传播。目前,蚜虫Hamiltonella和烟粉虱Hamiltonella的基因组测序已陆续完成,Hamiltonella基因组信息将为其功能研究提供新的思路与视角。     

MEAM1和MED两个烟粉虱隐种取食对甘蓝转录组影响的差异

为绿色防控烟粉虱Bemisia tabaci提供新策略,采用转录组分析MEAM1和MED两个烟粉虱隐种取食后甘蓝差异基因表达情况,并选取差异表达基因最多的甘蓝苯丙烷类通路上PAL2、C4H、4CL1和CHI四个基因进行实时荧光定量PCR测定。结果显示,与对照相比,MEAM1和MED烟粉虱隐种取食后甘蓝差异表达基因数分别为693个和1 030个。GO功能注释和KEGG富集分析显示,差异表达基因主要集中在苯丙烷类生物合成、萜类挥发物合成、芥子油苷合成、类黄酮生物合成和植物激素信号传导途径等与甘蓝抗虫物质合成相关的通路上。虽然MEAM1与MED两个烟粉虱隐种均能显著激活植物激素信号传导途径和苯丙烷类合成通路调控的酚类物质合成途径,但MED烟粉虱隐种与甘蓝互作的差异表达基因数高于MEAM1烟粉虱隐种。MEAM1烟粉虱隐种取食甘蓝12 h后,甘蓝苯丙烷通路上游PAL2、C4H和4CL1三个基因均显著上调,而MED烟粉虱隐种取食后,甘蓝苯丙烷通路上PAL2、C4H、4CL1和CHI四个基因均变化不显著,这4个差异基因的表达模式与测序结果一致,表明测序结果的可信度高。     

木尔坦棉花曲叶病毒在新疆的发生分布及烟粉虱带毒检测

为了明确木尔坦棉花曲叶病毒cotton leaf curl Multan virus (CLCuMuV)在新疆的发生分布,对采自新疆13个地(州)的1 166份蜀葵、苘麻、棉花疑似病株进行CLCuMuV分子鉴定和带毒检测,并对3种寄主上同期采集的1 588头烟粉虱进行隐种鉴定和带毒检测。结果显示,CLCuMuV DNA-A组分及其伴随的DNAβ卫星分子均可侵染蜀葵、苘麻、棉花;基于DNA-A及DNAβ组分的进化分析表明,新疆3种寄主上发生的CLCuMuV与国内已报道的CLCuMuV进化关系一致;蜀葵、苘麻、棉花上CLCuMuV带毒率存在差异,分别为27.87%、33.33%和14.95%;MEAM1和MED隐种为新疆烟粉虱主要种群,分别占26.70%和66.44%,带毒率分别为21.46%和17.63%,说明MED隐种为新疆烟粉虱优势种群,2个隐种均可携带CLCuMuV且带毒率相近;MEAM1隐种在蜀葵、苘麻、棉花上带毒率分别为21.39%、0和28.21%,MED隐种分别为19.61%,8.47%和11.18%,表明2个隐种在不同寄主上的带毒率存在差异。本研究明确了新疆地区侵染蜀葵、...     

温度胁迫下烟粉虱MED隐种与亚洲Ⅱ3隐种三个 抗寒基因表达模式的分析

烟粉虱Bemisia tabaci具有强的温度耐受性,但烟粉虱入侵种MED隐种与本地种亚洲Ⅱ3隐种在我国的地理分布上存在巨大的差异。因此本研究以烟粉虱MED隐种和亚洲Ⅱ3隐种为研究对象,采用qRT-PCR技术分别检测两种烟粉虱3个抗寒基因(erythrocyte binding protein,trehalase,dopadecarboxylase)在5个梯度温度胁迫下mRNA水平上的相对表达量,分析其在高低温胁迫过程中表达量的变化,探索抗寒基因在烟粉虱MED隐种成功入侵并广泛分布于各气候区域中所起的作用。研究结果表明这两种烟粉虱隐种体内,3个抗寒基因均显著受低温诱导。但相对于AsiaⅡ3隐种,低温胁迫下烟粉虱MED隐种体内的3种抗寒基因能更快速地做出响应,且基因上调或下调的量也相对较多。     

辽宁省冬季温室烟粉虱隐种鉴定

烟粉虱[Bemisia tabaci(Gennadius)]为辽宁省重要温室害虫,包括MEAM1隐种与MED隐种.为了揭示近年来辽宁省冬季温室内烟粉虱隐种动态,采用线粒体细胞色素氧化酶Ⅰ(mtCOI)PCR-RFLP技术,鉴别了辽宁省9个地区13个烟粉虱种群的隐种.结果表明,2019年冬季辽宁各地温室烟粉虱种群有12个...     

烟粉虱内共生菌传毒相关groEL基因的克隆及其原核表达

 烟粉虱内共生菌groEL基因编码一种63kD的GroEL蛋白,利用一对特异性引物,通过PCR对长期培养在黄瓜上的烟粉虱体内groEL基因进行了扩增,序列测定表明其长度为1668bp,编码555个氨基酸,与GenBank中烟粉虱内共生菌groEL(AF130421)的核苷酸同源性为99.58%,仅7个核苷酸发生了变异,二者氨基酸序列同源性为99.28%,仅4个氨基酸有别。构建了一个原核表达载体,表达得到了76kD的融合蛋白     

Secondary endosymbionts of Turkish Bemisia tabaci (Gennadius) populations

Secondary endosymbionts of Turkish Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations were determined by PCR-based DNA analysis. Experiments were conducted with B. tabaci samples collected from various host plants between 2007 and 2012. Four secondary endosymbionts, namely, Rickettsia, Hamiltonella, Arsenophonus and Wolbachia, were detected from two different B. tabaci species (B and Q). While Arsenophonus and Wolbachia were determined only from the Q, Hamiltonella was found only on the B. Rickettsia was determined on both B and Q. Forty percent of individuals were infected with Arsenophonus, followed by Hamiltonella (32.4 %), and Wolbachia (8 %). Infection rate of Rickettsia was found to be higher on B (29.7 %) than on Q (21.6 %). This study is the first report of endosymbionts of B. tabaci populations collected from Turkey, and studies should be continued to cover larger areas, more host plants and B. tabaci populations.     

Microsatellites reveal widespread predominance of an invasive over an indigenous <Emphasis Type="Italic">Bemisia tabaci</Emphasis> in Venezuela

The whitefly Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) is an important pest due to its capacity for producing strong infestations and transmitting plant viruses. The New World putative species of B. tabaci complex (NW) is the indigenous whitefly of the Americas, but only the invasive Middle East-Asia Minor 1 putative species of B. tabaci complex (MEAM1), commonly referred to as the “B biotype” was identified in a limited scope sampling in Venezuela. Similarly to MEAM1 invasions elsewhere, in this South American country there has been an increase in the geographic range and abundance of B. tabaci, and in the number of viruses that it transmits since the late 1980s. We estimated the diversity of B. tabaci to elucidate their role in the epidemiology of geminiviruses in Venezuela. Thirteen microsatellite loci were screened in samples collected from 19 localities in ten major agricultural states. A Bayesian clustering method (Structure) grouped the samples into two genetic groups. Control samples from whiteflies NW and MEAM1 and partial sequencing of the mitochondrial cytochrome oxidase I gene showed that our samples of B. tabaci populations from Venezuela fall within NW and MEAM1 groups. In this survey, MEAM1 was predominant over NW whitefly in a proportion of 35:1. No evidence was observed for gene flow between indigenous and invader whitefly. Altogether, our results stress the urgency for controlling the proliferation of the invasive whitefly.     

Panel of real‐time PCRs for the multiplexed detection of two tomato‐infecting begomoviruses and their cognate whitefly vector species

A new approach for the simultaneous identification of the viruses and vectors responsible for tomato yellow leaf curl disease (TYLCD) epidemics is presented. A panel of quantitative multiplexed real‐time PCR assays was developed for the sensitive and reliable detection of Tomato yellow leaf curl virus‐Israel (TYLCV‐IL), Tomato leaf curl virus (ToLCV), Bemisia tabaci Middle East Asia Minor 1 species (MEAM1, B biotype) and B. tabaci Mediterranean species (MED, Q biotype) from either plant or whitefly samples. For quality‐assurance purposes, two internal control assays were included in the assay panel for the co‐amplification of solanaceous plant DNA or B. tabaci DNA. All assays were shown to be specific and reproducible. The multiplexed assays were able to reliably detect as few as 10 plasmid copies of TYLCV‐IL, 100 plasmid copies of ToLCV, 500 fg B. tabaci MEAM1 and 300 fg B. tabaci MED DNA. Evaluated methods for routine testing of field‐collected whiteflies are presented, including protocols for processing B. tabaci captured on yellow sticky traps and for bulking of multiple B. tabaci individuals prior to DNA extraction. This work assembles all of the essential features of a validated and quality‐assured diagnostic method for the identification and discrimination of tomato‐infecting begomovirus and B. tabaci vector species in Australia. This flexible panel of assays will facilitate improved quarantine, biosecurity and disease‐management programmes both in Australia and worldwide.     

烟粉虱寄生蜂种类及繁殖方式多样性

本文列举了烟粉虱Bemisia tabaci寄生蜂种类,共计74种,其中恩蚜小蜂属Encarsia 47种,桨角蚜小蜂属Eretmocerus24种,埃宓细蜂属Amitus 2种及新姬小蜂属Neochrysocharis 1种;从生殖方式、寄生行为及寄主发育阶段喜好程度等方面论述了烟粉虱寄生蜂种类及繁殖方式的多样性;介绍了目前烟粉虱寄生蜂的引种工作;最后提出了今后有关烟粉虱寄生蜂的研究方向.     

Interactive effects of host plant and insecticide foliar application on oviposition and performance of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) cryptic species Mediterranean (MED) and Middle East-Asia minor 1 (MEAM1) in Brazil

Phytoparasitica - The Bemisia tabaci (Gennadius) cryptic species complex is composed of important pest insects, with cryptic species Mediterranean (MED) and Middle East-Asia Minor 1 (MEAM1) having...     

寄主植物对B型烟粉虱个体发育和种群繁殖的影响

同一原始种群B型烟粉虱[Bemisia tabaci(Gennadius)],分别在甘蓝、一品红、棉花、番茄和黄瓜连续隔离饲养大约20代后形成5种对应的烟粉虱种群,研究其在甘蓝上各虫态大小、存活率、发育历期、成虫寿命和平均产卵量等生物学参数。在(25±1)℃条件下,一品红种群烟粉虱的1、2、3龄若虫体长和宽均显著低于其他4个种群烟粉虱;发育时间以甘蓝和一品红种群烟粉虱(20.44、19.69d)显著高于番茄种群烟粉虱(19.13d);雌虫寿命以甘蓝种群烟粉虱显著低于一品红、棉花、番茄和黄瓜种群烟粉虱;平均单雌产卵量以一品红种群烟粉虱(169.30粒)显著高于其他4个种群烟粉虱;5个种群烟粉虱在甘蓝上的总存活率均无显著差异。     

The presence of Rickettsia is associated with increased susceptibility of Bemisia tabaci (Homoptera: Aleyrodidae) to insecticides

BACKGROUND: The presence of certain symbiotic microorganisms may be associated with insecticide resistance in insects. The authors compared the susceptibility of two isofemale lines, Rickettsia-plus and Rickettsia-free, of the sweet potato whitefly Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) to major insecticides from different chemical groups, including imidacloprid, acetamiprid, thiamethoxam, pyriproxyfen, spiromesifen and diafenthiuron. RESULTS: While the Rickettsia-plus and Rickettsia-free lines showed no differences in their susceptibility to imidacloprid and diafenthiuron, higher susceptibility of the Rickettsia-plus line to acetamiprid, thiamethoxam, spiromesifen and especially pyriproxyfen was observed. LC(90) values indicated that the Rickettsia-free line was 15-fold more resistant to pyriproxyfen than the Rickettsia-plus line. CONCLUSION: Findings indicate that the infection status of B. tabaci populations by Rickettsia is an important consideration that should be taken into account when performing resistance monitoring studies, and may help in understanding the dynamics of B. tabaci resistance, symbiont-pest associations in agricultural systems and the biological impact of Rickettsia on whitefly biology.     

红星盘瓢虫取食烟粉虱和桃蚜的生物学特性研究

本文研究了取食烟粉虱Bemisia tabaci（Gennadius）和桃蚜Myzus persicae（Sulzer）对红星盘瓢虫Phrynocaria congener（Billberg）生物学特性的影响,结果表明,取食不同虫态的烟粉虱时,红星盘瓢虫从卵到成虫不同虫态的发育历期和存活率差异不显著;取食不同虫态的桃蚜时,红星盘瓢虫一、三和四龄幼虫的发育历期和四龄幼虫的成活率差异显著。分别取食烟粉虱的混合种群（卵、各龄若虫和蛹）和桃蚜的混合种群（卵和各龄若虫）时,瓢虫从卵到成虫的发育历期和存活率差异显著,分别为13.3d和15.7d、81.1%和70.0%。红星盘瓢虫取食烟粉虱和桃蚜的产卵前期差异不显著;但成虫寿命和平均单雌产卵量差异显著,分别为64.6d和86.0d、389.4粒和960.4粒。红星盘瓢虫捕食烟粉虱和桃蚜的内禀增长率（rm）分别为0.0967和0.0964;净增殖率（R0）分别为118.7573和355.3586;世代平均历期（T）分别为49.4d和60.9d;周限增长率（λ）分别为1.1015和1.1012。     

Q型烟粉虱细胞色素P450 CYP6DV5基因克隆及其在烟粉虱对噻虫嗪抗性中的作用

烟粉虱Bemisia tabaci已严重危害农作物的正常生长,而新烟碱类杀虫剂噻虫嗪已被广泛用于烟粉虱的防治,但由于常年应用,致使烟粉虱对噻虫嗪产生了严重的抗性,而目前其抗药性分子机制尚不明确.本研究通过荧光定量PCR比较分析烟粉虱噻虫嗪抗性及敏感种群,发现细胞色素P450基因CYP6DV5在噻虫嗪抗性品系中的表达量为...