烟粉虱的分类地位及在中国的分布

烟粉虱广泛分布于全球热带和亚热带地区。近20多年,烟粉虱的一些遗传群入侵世界各地,严重危害作物生产。烟粉虱遗传结构的多样性和复杂性早已被关注,但其分类地位,尤其是烟粉虱到底是一个包含多个生物型的种还是一个包含许多隐种的物种复合体,一直颇受争议。近几年,有关烟粉虱种系发生和系统学的研究取得长足进展,有证据推论其是一个包含至少31个隐种的物种复合体,但生殖隔离证据仍显不足,种系发生分析结果也因仅依据COI一个基因而受到质疑。因此,在大多数从事烟粉虱研究的同行接受其为一个物种复合体的概念的同时,仍有同行沿用生物型的概念。在我国境内已先后报道了包括13个本地种和2个全球入侵种在内的15个烟粉虱隐种。本地种主要分布在我国南部及包括海南岛和台湾岛的东南沿海地区,隐种的多样性由南向北逐渐降低。入侵种“中东一小亚细亚1”隐种（MEAMl）（即“B型”）和“地中海”隐种（MED）（即“Q型”）分别于20世纪90年代中后期和2003年前后入侵我国,并在许多地区迅速取代了本地种而占据优势地位。全国范围内的调查数据显示,这2个入侵种可在大部分区域共同存在,但自2005年以来,MED在许多地区陆续取代MEAMl,这很可能与MED对大量使用的新烟碱类杀虫剂有较强抗性有关。本文还讨论了烟粉虱隐种复合体分类所面临的命名等难题以及大范围抽样调查的数据偏差问题。     

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.     

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.     

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。     

Genetic mutations associated with chemical resistance in the cytochrome P450 genes of invasive and native Bemisia tabaci (Hemiptera: Aleyrodidae) populations in China

Abstract  Bemisia tabaci (Gennadius) is a species complex, and its two most damaging biotypes B and Q are globally distributed pests. Despite increasing biological and economic impacts, little is known about the evolutionary mechanisms that favor their competition with native populations. Here, we investigated the genetic mutations in the P450 gene of the invasive B, Q biotypes and the native Cv population. Four mutations associated with chemical resistance, Pro-Leu, Ala-Ser, Ser-Phe and Trp-Leu, were found in the cytochrome P450 CYP6C and CYP9F genes of the B and Q biotypes. Bioassay results also revealed that both the B and Q biotypes have about 12–47 times more resistance to acephate, beta-cypermethrin, methomyl, and 5–7 times more resistance to imidacloprid insecticide than Cv population. Our results provide a molecular approach for better understanding and monitoring the pesticide resistances of invasive and native B. tabaci populations in China.     

福建省烟粉虱自然种群Wolbachia感染特点

Wolbachia是专性的细胞内细菌,广泛存在于节肢动物生殖组织。已有的研究结果表明,节肢动物中存在A组和B组Wolbachia,而烟粉虱Bemisia tabaci中主要检测到了B组Wolbachia。本研究从福建省采集到17个不同烟粉虱地理种群,首先通过rDNA-ITS1克隆测序鉴定了不同烟粉虱地理种群的生物型,然后采用Wolbachia 16S rDNA的特异引物,并通过PCR-RFLP技术分析了不同烟粉虱地理种群中Wolbachia的感染特点。结果表明：从福建省闽侯、平潭、南平、来舟、漳平和沙县采集到的烟粉虱自然种群属于非B型,而非B型烟粉虱种群中存在广泛的超感染现象,即单个非B型烟粉虱个体中同时感染了不同型Wolbachia。相反,B型烟粉虱自然种群的个体中只感染A组Wolbachia。该研究依据密集采样的数据进一步证实了Wolbachia在烟粉虱自然种群中的分布确实与宿主的生物型密切相关,提示Wolbachia可能在烟粉虱的种群分化中发挥作用。     

烟粉虱生物型研究进展

烟粉虱B em isia tabaci(G ennad ius)是一种取食植物汁液的重要农业害虫,同时也是许多植物病毒最重要的传播介体之一。烟粉虱被认为是由许多具有明显遗传分化的不同种群即生物型组成的复合种,其中B型烟粉虱是一种入侵性最强的生物型,几乎在世界各地均有分布。由于不同生物型的烟粉虱在寄主范围、传毒能力、地理分布、抗药性等许多生物学方面存在差异,因此对烟粉虱生物型的鉴定及其遗传分化研究对于该害虫的可持续控制具有重要的指导意义。烟粉虱生物型的鉴定方法包括生物学鉴定、酶谱鉴定以及分子标记鉴定的方法,其中使用的分子标记包括RAPD、AFLP、rDNA-ITS1、m tDNA CO I以及SSR等标记。目前,不同生物型的烟粉虱尤其B型烟粉虱的分类地位仍然存在争议。介绍了不同生物型烟粉虱的生物学差异、鉴定方法及其遗传分化研究的最新进展,同时探讨了不同生物型烟粉虱的分类地位和研究方向。     

华东、华南和西南地区烟粉虱的种群及寄主分化

烟粉虱Bemisia tabaci是一个复合种,它具有的生物型分化、较强的传播病毒的能力和抗药性、较快的繁殖速率等特征使其成为我国农业生产中重要害虫之一。本研究利用细胞色素线粒体氧化酶Ⅰ基因,对采集自江苏、广东和云南三省的烟粉虱样本进行了生物型鉴定,并对烟粉虱生物型与寄主植物之间的关联性开展了调查。结果表明,在广东和云南省,都存在未鉴定的土著种群与入侵的B型、Q型共存的现象;同时,在本研究中广东省尚未采集到Q型烟粉虱,而在江苏采集到的粉虱样本全部为入侵型。研究结果还表明,相对于入侵种而言,土著种群显示出更强的寄主植物趋同性;丰富的寄主植物以及本身具有的多食性特性有助于B型、Q型等生物型在世界各地的广泛入侵。     

辽宁省冬季温室内越冬粉虱伪蛹的种类鉴定及烟粉虱体内番茄黄化曲叶病毒的检测

为了揭示辽宁省冬季温室内越冬粉虱伪蛹的种类及烟粉虱Bemisia tabaci （Gennadius）携带番茄黄化曲叶病毒（tomato yellow leaf curl virus, TYLCV）情况, 于2012年1月份在辽宁省不同县市区的温室作物上采集了17份粉虱伪蛹样品（每样品含30头粉虱伪蛹） , 镜检鉴别粉虱种类并利用mtCOI基因对烟粉虱生物型进行了鉴定; 检测了烟粉虱携带TYLCV情况并对其PCR扩增产物进行了测序分析。结 果表明： 辽宁省冬季温室内存在越冬温室白粉虱Trialeurodes vaporariorum （Westwood）与烟粉虱。17份粉虱样品中, 11份样品为烟 粉虱样品, 6份样品为温室白粉虱和烟粉虱混合样品。混合样品中, 温室白粉虱仅在锦州凌海（LH）样品中占优势。17份烟粉虱样品（包 括混合样品）中, 仅有4份样品为B型与Q型混合样品, 其他13份样品烟粉虱生物型均为Q型。17份烟粉虱样品中有3份Q型烟粉虱样品检测 到TYLCV, 系统树分析进一步证实该病毒是TYLCV。调查结果为辽宁省粉虱与TYLCV的早期测报和防控提供了科学依据。     

The root-knot nematode resistance gene Mi-1.2 of tomato is responsible for resistance against the whitefly Bemisia tabaci

The tomato gene Mi-1.2 confers resistance against root-knot nematodes and some isolates of potato aphid. Resistance to the whitefly Bemisia tabaci previously has been observed in Mi-bearing commercial tomato cultivars, suggesting that Mi, or a closely linked gene, is responsible for the resistance. The response of two biotypes of B. tabaci to tomato carrying the cloned Mi was compared with that of the isogenic untransformed tomato line Moneymaker. Our results indicate that Mi-1.2 is responsible for the resistance in tomato plants to both B- and Q- biotypes. Mi-1.2 is unique among characterized resistance genes in its activity against three very different organisms (root-knot nematodes, aphids, and whiteflies). These pests are among the most important on tomato crops worldwide, making Mi a valuable resource in integrated pest management programs.     

Benzothiadiazole induces local resistance to Bemisia tabaci (Hemiptera: Aleyrodidae) in tomato plants

Plant resistance to the B and Q biotypes of sweetpotato whitefly, Bemisa tabaci (Gennadius), induced by benzo [1,2,3] thiadiazole-7-carbothioic acid-S-methyl ester (BTH or acibenzolar-S-methyl) in tomato 'Marmande' plants was evaluated in free-choice and no-choice assays under different conditions. BTH is the active ingredient of the Syngenta plant activator Bion. BTH treatment affected host preference of B. tabaci (B and Q biotypes) adults on plants sprayed with Bion at 0.2 and 0.4 g/liter during the earlier days of free-choice assays. As a consequence, a decrease in the total number of eggs (although female fecundity was not affected) and in the final number of pupae and empty pupal cases was observed. The effect produced by BTH applied at 0.1 g/liter Bion was not significant. In no-choice assays, a reduction of the numbers of first-stage larvae and total individuals and a delay in insect development were observed when local treatment was restricted to one leaflet per plant, 5 d before B. tabaci (biotype B) infestation. This acquired resistance induced by BTH seemed to be locally expressed because of the differences between treated and nontreated leaflets in the same plants, whereas no differences in nontreated leaflets were observed between BTH-treated and control plants.     

Lack of fitness costs associated with acetamiprid resistance in Bemisia tabaci (Hemiptera: Aleyrodidae)

Sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is a devastating pest that can cause severe damage to a range of crops by direct feeding and by plant virus transmission. Because of indiscriminate use of insecticides, this whitefly has developed resistance to several insecticides, including neonicotinoids. Our objectives were to determine fitness components affected by acetamiprid resistance in B. tabaci. Assay results showed that selection with acetamiprid had removed heterozygotes from the field population because the survival rate of the resistant population was significantly greater than that of the field population at a very high dose. Comparison of various life traits between the acetamiprid-selected (Aceta-SEL) population and three other populations showed that the numbers of eggs laid by acetamiprid Aceta-SEL population were significantly lower compared with that of other populations but that the proportions of eggs hatched were significantly higher. However, the time taken by nymphal stages of the Aceta-SEL population to develop was significantly higher than that of the susceptible populations. The intrinsic rate of increase, net reproductive rate, mean generation time, and doubling time of Aceta-SEL was significantly higher than Lab-PK and UNSEL populations, but the growth index was similar for all populations. The growth index and high intrinsic value of Aceta-SEL population suggest that the resistance allele may not have detrimental impact. The lack of fitness costs in B. tabaci could promote the rapid development of resistance to acetamiprid and other neonicotinoids. This resistance could threaten the sustainability of whitefly management program on genetically engineered cotton (Gossypium hirsutum L.) where neonicotinoids are being sprayed to manage sucking pests in the field.     

Reproductive incompatibility between the B and Q biotypes of the whitefly Bemisia tabaci in China: genetic and behavioural evidence

The B and Q 'biotypes' of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) have been invading many parts of the world and causing severe damage to a range of crops. Recent phylogenetic analyses indicate that B and Q are cryptic species within the B. tabaci species complex. Although various attempts have been made to examine the reproductive compatibility between B and Q, few studies have tested the fertility of the F1 females and so the extent of possible gene flow remains unclear. In this study, we conducted a series of crossing experiments and behavioural observations to examine in detail the reproductive compatibility between the B and Q biotypes collected from Zhejiang, China, a region recently invaded by these whiteflies. Crossing experiments between the two biotypes using either single-pairs or small groups demonstrated that proportions of females in the F1 progeny were only 0-2% in the inter-biotype crosses compared to 58-68% in the intra-biotype treatments. Furthermore, all inter-biotype F1 females were sterile. Continuous video observations showed that B and Q adults very rarely copulated, and copulation occurred only when adults of opposite sex from different biotypes were enclosed in dense cohorts for a relatively long period of time. These data show that the B and Q biotypes examined in this study are completely isolated in reproduction. The isolation was due to mainly a copulation barrier, but post-copulation barriers were also involved.     

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。     

Wolbachia infections of the whitefly Bemisia tabaci

We report the first systematic survey for the presence of Wolbachia endosymbionts in aphids and whiteflies, particularly different populations and biotypes of Bemisia tabaci. Additional agriculturally important species included were predator species, leafhoppers, and lepidopterans. We used a polymerase chain reaction (PCR)-based detection assay with ribosomal 16S rDNA and Wolbachia cell surface protein (wsp) gene primers. Wolbachia were detected in a number of whitefly populations and species, whitefly predators, and one leafhopper species; however, none of the aphid species tested were found infected. Single, double, and triple infections were detected in some of the B. tabaci populations. PCR and phylogenetic analysis of wsp gene sequences indicated that all Wolbachia strains found belong to group B. Topologies of the optimal tree derived by maximum likelihood (ML) and a ML tree in which Wolbachia sequences from B. tabaci are constrained to be monophyletic are significantly different. Our results indicate that there have been at least four independent Wolbachia infection events in B. tabaci. The importance of the presence of Wolbachia infections in B. tabaci is discussed.     

Use of RAPD PCR to Distinguish the B Biotype from Other Biotypes of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae)

The use of RAPD PCR to identify the B biotype of the whitefly Bemisia tabaci and distinguish it from other biotypes and species of whitefly is described. the technique enables the use of alcohol preserved material instead of live or frozen material as required by allozyme electrophoresis and demonstrates that eggs, juvenile stages and males or females can all be used.     

Over-expression of cytochrome P450 CYP6CM1 is associated with high resistance to imidacloprid in the B and Q biotypes of Bemisia tabaci (Hemiptera: Aleyrodidae)

The two most damaging biotypes of Bemisia tabaci, B and Q, have both evolved strong resistance to the neonicotinoid insecticide imidacloprid. The major mechanism in all samples investigated so far appeared to be enhanced detoxification by cytochrome P450s monooxygenases (P450s). In this study, a polymerase chain reaction (PCR) technology using degenerate primers based on conserved P450 helix I and heme-binding regions was employed to identify P450 cDNA sequences in B. tabaci that might be involved in imidacloprid resistance. Eleven distinct P450 cDNA sequences were isolated and classified as members of the CYP4 or CYP6 families. The mRNA expression levels of all 11 genes were compared by real-time quantitative RT-PCR across nine B and Q field-derived strains of B. tabaci showing strong resistance, moderate resistance or susceptibility to imidacloprid. We found that constitutive over-expression (up to approximately 17-fold) of a single P450 gene, CYP6CM1, was tightly related to imidacloprid resistance in both the B and Q biotypes. Next, we identified three single-nucleotide polymorphic (SNP) markers in the intron region of CYP6CM1 that discriminate between the resistant and susceptible Q-biotype CYP6CM1 alleles (r-Q and s-Q, respectively), and used a heterogeneous strain to test for association between r-Q and resistance. While survivors of a low imidacloprid dose carried both the r-Q and s-Q alleles, approximately 95% of the survivors of a high imidacloprid dose carried only the r-Q allele. Together with previous evidence, the results reported here identify enhanced activity of P450s as the major mechanism of imidacloprid resistance in B. tabaci, and the CYP6CM1 gene as a leading target for DNA-based screening for resistance to imidacloprid and possibly other neonicotinoids in field populations.     

Asymmetrical crossing barriers in angiosperms

Patterns of reproductive isolation between species may provide insight into the mechanisms and evolution of barriers to interspecific gene exchange. We used data from published interspecific hybridization experiments from 14 genera of angiosperms in order to test for the presence of asymmetrical barriers to gene exchange. Reproductive isolation was examined at three life-history stages: the ability of interspecific crosses to produce seeds, the viability of F1 hybrids, and the fertility of F1 hybrids. Statistically significant asymmetries in the strength of reproductive isolation between species were detected in all genera and at each of the three life-history stages. Asymmetries in seed production may be caused by a variety of mechanisms including differences in stigma/style lengths, self compatibility and differential fruit abortion. Asymmetries in post-zygotic isolation are probably caused by nuclear-cytoplasmic interactions. Asymmetrical reproductive isolation between plant taxa may have important implications for the dynamics of hybrid zones, the direction of genetic introgression and the probability of reinforcement.     

Differential indirect effects of two plant viruses on an invasive and an indigenous whitefly vector: implications for competitive displacement

The role of vector–begomovirus–plant interactions in the widespread invasion by some members of the whitefly species complex Bemisia tabaci is poorly understood. The invasive B biotype of B. tabaci entered China in the late 1990s and had become the predominant or only biotype of the whitefly in many regions of the country by 2005–2006. Meanwhile epidemics of begomoviruses have been observed in many crops including tomato for which Tomato yellow leaf curl China virus (TYLCCNV) and Tomato yellow leaf curl virus (TYLCV) have been identified as two major disease-causing agents. Here, we conducted laboratory experiments to compare the performance of the invasive B and indigenous ZHJ1 whitefly biotypes on uninfected, TYLCCNV-infected and TYLCV-infected plants of tomato cv. Hezuo903, a cultivar that has been widely cultivated in many regions of China. The infection of tomato plants by either of the viruses had no or only marginal effects on the development, survival and fecundity of the B biotype. In contrast, survival and fecundity of the ZHJ1 biotype were significantly reduced on virus-infected plants compared to those on uninfected plants. Populations of the B biotype on uninfected and TYLCCNV-infected plants increased at similar rates, whereas population increase of the ZHJ1 biotype on TYLCCNV-infected plants was affected adversely. These asymmetric responses to virus infection of tomato plants between the B and ZHJ1 biotypes are likely to offer advantages to the B biotype in its invasion and displacement of the indigenous biotype.     

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。