Cardinium symbionts cause cytoplasmic incompatibility in spider mites

Intracellular symbiotic bacteria belonging to the Cytophaga-Flavobacterium-Bacteroides lineage have recently been described and are widely distributed in arthropod species. The newly discovered bacteria, named Cardinium sp, cause the expression of various reproductive alterations in their arthropod hosts, including cytoplasmic incompatibility (CI), induction of parthenogenesis and feminization of genetic males. We detected 16S ribosomal DNA sequences similar to those of Cardinium from seven populations of five spider mite species, suggesting a broad distribution of infection of Cardinium in spider mites. To clarify the effect of Cardinium on the reproductive traits of the infected spider mites, infected mites were crossed with uninfected mites for each population. In one of the populations, Eotetranychus suginamensis, CI was induced when infected males were crossed with uninfected females. The other six populations of four species showed no reproductive abnormalities in the F(1) generation, but the possibility of CI effects in the F(2) generation remains to be tested. One species of spider mite, Tetranychus pueraricola, harbored both Cardinium and Wolbachia, but these symbionts seemed to have no effect on the reproduction of the host, even when the host was infected independently with each symbiont.     

Distribution of the bacterial symbiont Cardinium in arthropods

Abstract 'Candidatus Cardinium', a recently described bacterium from the Bacteroidetes group, is involved in diverse reproduction alterations of its arthropod hosts, including cytoplasmic incompatibility, parthenogenesis and feminization. To estimate the incidence rate of Cardinium and explore the limits of its host range, 99 insect and mite species were screened, using primers designed to amplify a portion of Cardinium 16S ribosomal DNA (rDNA). These arthropods were also screened for the presence of the better-known reproductive manipulator, Wolbachia. Six per cent of the species screened tested positive for Cardinium, compared with 24% positive for Wolbachia. Of the 85 insects screened, Cardinium was found in four parasitic wasp species and one armoured scale insect. Of the 14 mite species examined, one predatory mite was found to carry the symbiont. A phylogenetic analysis of all known Cardinium 16S rDNA sequences shows that distantly related arthropods can harbour closely related symbionts, a pattern typical of horizontal transmission. However, closely related Cardinium were found to cluster among closely related hosts, suggesting host specialization and horizontal transmission among closely related hosts. Finally, the primers used revealed the presence of a second lineage of Bacteroidetes symbionts, not related to Cardinium, in two insect species. This second symbiont lineage is closely allied with other arthropod symbionts, such as Blattabacterium, the primary symbionts of cockroaches, and male-killing symbionts of ladybird beetles. The combined data suggest the presence of a diverse assemblage of arthropod-associated Bacteroidetes bacteria that are likely to strongly influence their hosts' biology.     

Factors affecting population dynamics of maternally transmitted endosymbionts in Bemisia tabaci

While every individual of Bemisia tabaci (Hemiptera: Aleyrodidae) harbors the primary symbiont (P-symbiont) Portiera, the infection frequencies of the six secondary symbionts (S-symbionts) including Hamiltonella, Arsenophonus, Cardinium, Wolbachia, Rickettsia and Fritschea vary greatly among different populations. To characterize the factors influencing the infection dynamics of the six S-symbionts in B. tabaci, gene-specific PCR were conducted to screen for the presence of the P-symbiont Portiera and the six S-symbionts in 61 (17 B and 44 Q biotypes) field populations collected from different plant species and locations in China. All individuals of the 61 populations hosted the P-symbiont Portiera, but none of them harbored Arsenophonus and Fritschea. The presence and infection rates of Hamiltonella, Cardinium, Rickettsia, Wolbachia and their co-infections Rickettsia + Hamiltonella (RH), Rickettsia + Cardinium (RC), Hamiltonella + Cardinium (HC) and Rickettsia + Hamiltonella + Cardinium (RHC) varied significantly among the 61 field populations; and the observed variations can be explained by biotypes, sexes, host plants and geographical locations of these field populations. Taken together, at least three factors including biotype, host plant and geographical location affect the infection dynamics of S-symbionts in B. tabaci.     

Diversity and recombination in <Emphasis Type="Italic">Wolbachia</Emphasis> and <Emphasis Type="Italic">Cardinium</Emphasis> from <Emphasis Type="Italic">Bryobia</Emphasis>spider mites

BACKGROUND: Wolbachia and Cardinium are endosymbiotic bacteria infecting many arthropods and manipulating host reproduction. Although these bacteria are maternally transmitted, incongruencies between phylogenies of host and parasite suggest an additional role for occasional horizontal transmission. Consistent with this view is the strong evidence for recombination in Wolbachia, although it is less clear to what extent recombination drives diversification within single host species and genera. Furthermore, little is known concerning the population structures of other insect endosymbionts which co-infect with Wolbachia, such as Cardinium. Here, we explore Wolbachia and Cardinium strain diversity within nine spider mite species (Tetranychidae) from 38 populations, and quantify the contribution of recombination compared to point mutation in generating Wolbachia diversity. RESULTS: We found a high level of genetic diversity for Wolbachia, with 36 unique strains detected (64 investigated mite individuals). Sequence data from four Wolbachia genes suggest that new alleles are 7.5 to 11 times more likely to be generated by recombination than point mutation. Consistent with previous reports on more diverse host samples, our data did not reveal evidence for co-evolution of Wolbachia with its host. Cardinium was less frequently found in the mites, but also showed a high level of diversity, with eight unique strains detected in 15 individuals on the basis of only two genes. A lack of congruence among host and Cardinium phylogenies was observed. CONCLUSIONS: We found a high rate of recombination for Wolbachia strains obtained from host species of the spider mite family Tetranychidae, comparable to rates found for horizontally transmitted bacteria. This suggests frequent horizontal transmission of Wolbachia and/or frequent horizontal transfer of single genes. Our findings strengthens earlier reports of recombination for Wolbachia, and shows that high recombination rates are also present on strains from a restrictive host range. Cardinium was found co-infecting several spider mite species, and phylogenetic comparisons suggest also horizontal transmission of Cardinium among hosts.     

Wolbachia和Cardinium对皮氏叶螨生殖的影响及在寄主体内的定位

Wolbachia和Cardinium均为母系遗传的胞内共生菌, 它们能够通过诱导胞质不亲和（cytoplasmic incompatibility, CI）以调控寄主的生殖。目前, 关于Wolbachia和Cardinium共同对同一寄主进行生殖操控的机制还不清楚。本研究以皮氏叶螨Tetranychus piercei McGregor广州种群为实验材料, 通过杂交实验和荧光原位杂交的方法, 研究Wolbachia和Cardinium单感染和双感染对寄主生殖的影响。结果表明： 单感染Wolbachia诱导较弱的CI, 不亲和组合的未孵化率为17.8%±1.6%。单感染Cardinium及双感染Wolbachia和Cardinium能诱导高强度的CI, 不亲和组合的未孵化率分别为70.3%±1.3%和72.9%±1.2%。同时双感染Wolbachia和Cardinium雌螨的平均产卵量为35.2±1.2, 显著高于单感染和不感染的雌螨的产卵量。Wolbachia 和Cardinium分别诱导以及共同诱导CI的水平与精子形成过程中的感染情况有关。Wolbachia和Cardinium的垂直传播模式结果显示, 在卵的不同发育阶段, Wolbachia和Cardinium主要伴随着营养物质从滋养细胞、 中肠、 输卵管进入发育中的卵。研究结果为进一步了解 Wolbachia和Cardinium的母系遗传机制提供了重要依据。     

显微注射青霉素G获得单感染Wolbachia和单感染Cardinium白背飞虱品系

Wolbachia和Cardinium都是广泛存在于节肢动物体内的一类母系遗传的共生细菌, 可以通过不同方式操纵寄主的生殖行为。Wolbachia和Cardinium感染同一寄主在自然界比较常见, 但是在某些可以同时感染Wolbachia和Cardinium的寄主中其单感染品系较难发现。本研究检测了云南文山(YN)、 海南三亚(HN)这2个不同地理种群中Wolbachia和Cardinium的感染情况; 以双感染Wolbachia和Cardinium的白背飞虱Sogatella furcifera海南种群为实验材料, 运用显微注射方法对双感染Wolbachia和Cardinium的白背飞虱若虫注射不同浓度青霉素G以获得单感染品系。结果表明： 白背飞虱自然种群中单感染Wolbachia比率极低, 本实验用到的海南种群未检测到单感染Wolbachia成虫; 通过显微注射青霉素G的方法可以从白背飞虱双感染品系中筛选获得单感染品系, 当青霉素G注射浓度为0.2%(w/v), 注射龄期为5龄时得到单感染品系效率最高; F5代的检测结果显示显微注射得到的单感染品系可以稳定遗传。本研究为获得单感染品系白背飞虱提供了快捷方法, 同时为其他双感染Wolbachia和Cardinium节肢动物不同感染品系的筛选提供参考。     

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.     

Cryptic diversity of the symbiotic cyanobacterium Synechococcus spongiarum among sponge hosts

Cyanobacteria are common members of sponge-associated bacterial communities and are particularly abundant symbionts of coral reef sponges. The unicellular cyanobacterium Synechococcus spongiarum is the most prevalent photosynthetic symbiont in marine sponges and inhabits taxonomically diverse hosts from tropical and temperate reefs worldwide. Despite the global distribution of S. spongiarum , molecular analyses report low levels of genetic divergence among 16S ribosomal RNA (rRNA) gene sequences from diverse sponge hosts, resulting either from the widespread dispersal ability of these symbionts or the low phylogenetic resolution of a conserved molecular marker. Partial 16S rRNA and entire 16S–23S rRNA internal transcribed spacer (ITS) genes were sequenced from cyanobacteria inhabiting 32 sponges (representing 18 species, six families and four orders) from six geographical regions. ITS phylogenies revealed 12 distinct clades of S. spongiarum that displayed 9% mean sequence divergence among clades and less than 1% sequence divergence within clades. Symbiont clades ranged in specificity from generalists to specialists, with most (10 of 12) clades detected in one or several closely related hosts. Although multiple symbiont clades inhabited some host sponges, symbiont communities appear to be structured by both geography and host phylogeny. In contrast, 16S rRNA sequences were highly conserved, exhibiting less than 1% sequence divergence among symbiont clades. ITS gene sequences displayed much higher variability than 16S rRNA sequences, highlighting the utility of ITS sequences in determining the genetic diversity and host specificity of S. spongiarum populations among reef sponges. The genetic diversity of S. spongiarum revealed by ITS sequences may be correlated with different physiological capabilities and environmental preferences that may generate variable host–symbiont interactions.     

Physiological performance of juvenile Haliotis rufescens and Haliotis discus hannai abalone exposed to the withering syndrome agent

The presence of Wolbachia and Cardinium bacteria has been documented in many arthropod species, including the predatory mite Metaseiulus (=Typhlodromus or Galendomus) occidentalis (Nesbitt) (Acari: Phytoseiidae). We show that Tetranychus urticae, the prey of Metaseiulus occidentalis, contains Wolbachia and no detectable Cardinium using quantitative PCR (qPCR). Starvation for 72 h at 22°C eliminated most, if not all, Wolbachia in M. occidentalis adult females from 7 laboratory colonies. Refeeding of M. occidentalis with T. urticae after starvation for 72 h restored the amounts of Wolbachia in M. occidentalis to those of prestarvation levels, suggesting that Wolbachia detected in M. occidentalis starved for shorter periods of time in current, and some previous, studies likely came from T. urticae. Furthermore, eggs from all M. occidentalis colonies examined were free of Wolbachia if they were surface-decontaminated with 0.3% sodium hypochlorite before DNA extraction. Cardinium was present in 6 of 14 laboratory colonies of M. occidentalis. Starvation for 3, 24, 48, and 72 h had no effect on the amounts of Cardinium in adult females from the Cardinium-positive colonies. Eggs from these colonies were positive for Cardinium but contained less than 1% of the titers found in adult females. The data suggest that Cardinium, but not Wolbachia, is an endosymbiont in certain populations of M. occidentalis. In light of our current findings, we recommend specific practices for the identification of potential symbionts in predatory arthropod species using the PCR.     

海南地区螺旋粉虱三类次级内共生菌的检测

螺旋粉虱Aleurodicus dispersus Russell是一种重要的农林害虫。本研究分别利用次级内共生菌Cardinium和Arsenophonus的16S rDNA和Wolbachia wsp基因对海南省16地区的螺旋粉虱的3种次级内共生菌Cardinium, Arsenophonus和Wolbachia感染情况及相关基因序列进行了测定和分析。3种次级内共生菌Cardinium, Arsenophonus和Wolbachia检测结果表明, Cardinium和Arsenophonus均可感染海南地区的螺旋粉虱, 其中乐东、 陵水和澄迈3个地区所有寄主上的螺旋粉虱的Arsenophonus感染率为100%, 三亚、 琼中和临高部分寄主上的螺旋粉虱的Arsenophonus感染率为66.7%, 而儋州、 五指山和万宁3个地区的螺旋粉虱未发现被Arsenophonus感染; 三亚的番石榴上的螺旋粉虱的Cardinium感染率为100%, 琼海白沙的印度紫檀上螺旋粉虱的Cardinium感染率为100%, 其他寄主上的感染率均小于66.7%; 在所检测的43个螺旋粉虱种群中, 40和31个种群中分别检测出有Arsenophonus 和Cardinium感染, 种群感染率分别为93.0%和72.1%; 在所有检测的个体中, 120个个体中有105个感染Arsenophonus, 93个个体中有70个感染Cardinium, 个体感染率分别为87.5% 和75.3%; 在检测的所有样本中, 只有三亚印度紫檀上的螺旋粉虱检测到Wolbachia, 种群感染率为2.3%, 个体感染率仅为0.8%。这些检测结果表明, 海南地区螺旋粉虱次级内共生菌Arsenophonus的感染率高于Cardinium的感染率, Wolbachia的感染率极低。序列分析表明, 海南不同螺旋粉虱种群的Cardinium的16S rDNA序列一致, 而且与烟粉虱的Cardinium 16S rDNA序列一致性很高, 为97.6%; 不同螺旋粉虱种群的Arsenophonus的16S rDNA序列也完全一致, 其与西班牙加那利群岛螺旋粉虱的Arsenophonus的16S rDNA序列一致性较高, 为85.1%。此外, Wolbachia wsp基因序列分析表明, 海南螺旋粉虱的Wolbachia为B组, 这是国内螺旋粉虱感染Wolbachia的首次报道。     

铃木氏果蝇不同地理种群中Wolbachia的检测和系统发育分析

铃木氏果蝇Drosophila suzukii是原产于东南亚地区的重要果树害虫, 近年来传入北美和欧洲等地区造成严重的危害。本研究利用Wolbachia的16S rDNA和wsp基因特异引物（分别为16S-F/16S-R和81F/691R）对铃木氏果蝇7个地理种群（中国的5个种群、 韩国的1个种群和美国的1个种群）的Wolbachia进行了PCR检测并对检测结果进行了比较; 对感染个体体内Wolbachia的16S rDNA基因片段进行测序, 确定了我国铃木氏果蝇体内Wolbachia的分类地位。基于Wolbachia的16S rDNA基因特异引物检测结果发现, 我国5个铃木氏果蝇种群广泛感染Wolbachia（感染率36.7%~80.0%）, 而韩国和美国2个种群均未检测到该菌的感染。而利用wsp基因特异引物无法检测到该菌。基于Wolbachia的16S rDNA基因构建系统发育树表明, 我国铃木氏果蝇种群感染的Wolbachia全部属于A组。这些结果为研究Wolbachia感染对铃木氏果蝇生物学及生态学的影响奠定了基础。     

Wolbachia在我国不同地理种群的玉米螟赤眼蜂中的分布

Wolbachia是一类可以引起寄主生殖行为改变的共生菌,广泛存在于节肢动物内。玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen是我国亚洲玉米螟Ostrinia furnacalis(Guene)卵寄生蜂的优势种。本文利用Wolbachia的16SrDNA基因特异性引物进行PCR扩增以及克隆测序,对我国14个地区的玉米螟赤眼蜂种群中Wolbachia的感染情况做了详细的调查。结果发现,被测的1412头野生玉米螟赤眼蜂全部感染有Wolbachia。系统发育分析表明,北京柳沟、山东菏泽、河南郑州、河南驻马店、湖南长沙5个种群存在超感染现象。这是Wolbachia在玉米螟赤眼蜂种群内分布情况的首次报道。     

Cardinium is associated with reproductive incompatibility in the predatory mite Metaseiulus occidentalis (Acari: Phytoseiidae)

Cardinium, a bacterium from the Bacteroidetes group, is associated with reproductive manipulations such as cytoplasmic incompatibility, parthenogenesis, and feminization in some arthropod species. We, and others, have shown that Cardinium, but not Wolbachia, is an endosymbiont in some populations of Metaseiulus occidentalis, a phytoseiid mite that is an important predator of spider mite pests of agricultural crops. However, the precise biological effects that Cardinium may have on M. occidentalis remain unclear. In this study we show, in two sets of crosses between different colonies of Cardinium-free (C-) M. occidentalis females and Cardinium-containing (C+) males, that fecundity was reduced in parental females, F1 progeny survival rates were reduced, and fewer female progeny were produced when compared to the reciprocal and control crosses. There were no differences in these attributes in the reciprocal and two control crosses. Cardinium was transmitted maternally but there was no observed paternal transmission. Finally, Cardinium did not cause asexual (thelytoky) reproduction in M. occidentalis. Thus, Cardinium is associated with nonreciprocal reproductive incompatibility in M. occidentalis and our results support the hypothesis that Cardinium is a reproductive parasite in this agriculturally important predator. Cardinium may therefore affect the evolution and ecology of M. occidentalis and biological control efforts using this mite.     

The microbiota in spider mite feces potentially reflects intestinal bacterial communities in the host

Microorganisms provide many physiological functions to herbivorous hosts. Spider mites (genus Tetranychus) are important agricultural pests throughout the world; however, the composition of the spider mite microbial community, especially gut microbiome, remains unclear. Here, we investigated the bacterial community in five spider mite species and their associated feces by deep sequencing of the 16S rRNA gene. The composition of the bacterial community was significantly different among the five prevalent spider mite species, and some bacterial symbionts showed host‐species specificity. Moreover, the abundance of the bacterial community in spider mite feces was significantly higher than that in the corresponding spider mite samples. However, Flavobacterium was detected in all samples, and represent a “core microbiome”. Remarkably, the maternally inherited endosymbiont Wolbachia was detected in both spider mite and feces. Overall, these results offer insight into the complex community of symbionts in spider mites, and give a new direction for future studies.     

High incidence of the maternally inherited bacterium Cardinium in spiders

Inherited bacteria are now recognized as important players in arthropod evolution and ecology. Here, we test spiders, a group recently identified as possessing inherited bacteria commonly, for the presence of two reproductive parasites, Cardinium hertigii (Bacteroidetes group) and Wolbachia (α-proteobacteria), estimating incidence, prevalence, any sex bias in infection, and infection diversity, for a panel of field-collected specimens. We identify spiders as a hotspot for Cardinium . Present in 22% of the sampled species, incidence was significantly higher than that previously recorded in insects. Where present, Cardinium infection occurred at medium prevalence without evidence of sex bias in prevalence that would indicate sex-ratio distortion activity. Wolbachia was present in 37% of species, but revealed a gradation from being rare to very common. In one case, Wolbachia was found significantly more commonly in females than males, indicating it may act as a sex-ratio distorter in some species. Breeding work conducted on two species confirmed that Wolbachia and Cardinium were transmitted maternally, which represents the first proof of inheritance of these symbionts in spiders. Overall, this study demonstrates that the majority of spider species are infected with inherited bacteria, and their role in host biology clearly requires determination.     

Geographical structuring of feather mite assemblages from the Australian brush-turkey (Aves: Megapodiidae)

Populations of a host species may exhibit different assemblages of parasites and other symbionts. The loss of certain species of symbionts (lineage sorting, or "missing-the-boat") is a mechanism by which geographical variation in symbiont assemblages can arise. We studied feather mites and lice from Australian brush-turkeys (Aves: Megapodiidae: Alectura lathami) and expected to observe geographical structuring in arthropod assemblages for several reasons. First, because the brush-turkey is a sedentary ground-dwelling bird, we predicted that geographically close host populations should share more similar arthropod assemblages than distant ones. Second, because brush-turkeys do not brood their young, vertical transfer of arthropods is unlikely, and brush-turkeys probably acquire their mites and lice at social maturity through contact with other birds. Young birds could disperse and found new populations without carrying complete sets of symbionts. We predicted that young birds would have fewer species of arthropods than older birds; in addition, we expected that males (which are polygynous) would have more species than females. Birds were sampled from 12 sites (=populations) along the east coast of Queensland, Australia, that were separated by a distance of 12.5-2,005 km. In total, 5 species of mites from the Pterolichidae and 1 species from the Ascouracaridae were found. Two species of lice were collected but in numbers too low to be statistically useful. Differentiation of mite assemblages was evident; in particular, Leipobius sp. showed 100% prevalence in 3 host populations and 0% in the remaining 9. A dendrogram of brush-turkey populations based on mite assemblages showed 2 geographically correlated clusters of sites, plus 1 cluster that contained 2 sites near Brisbane and 1 approximately at a distance of 1,000 km. There was no strong effect of host age or sex on number of mite species carried. Horizontal transfer of feather mites by hippoboscid flies, in addition to physical contact between hosts, may play a role in homogenizing symbiont assemblages within populations.     

白背飞虱中的Wolbachia和Cardinium双重感染特性

为了明确自然种群白背飞虱Sogatella furcifera中Wolbachia和Cardinium的感染情况以及Wolbachia与其特有的WO噬菌体之间的关系, 以采自中国7个省区9个地点的白背飞虱为研究材料, 运用PCR检测的方法调查了Wolbachia, Cardinium以及WO噬菌体在各飞虱种群中的感染率和组织分布特点。结果表明： 白背飞虱广泛双重感染Wolbachia和Cardinium, 并且都表现出很高的感染率。白背飞虱各种群Cardinium的感染率几乎均为100%; Wolbachia的感染率也较高, 但雌雄虫感染率差异较大, 雌虫的感染率几乎均为100％, 而雄虫的感染率从22.2%~95.0%不等。另外, 通过不同DNA聚合酶、 不同提取方法的对比, 揭示了DNA粗提样品在基于PCR技术的胞内共生菌检测中的不足之处。对白背飞虱头部、 胸部、 腹部、 足和翅5个不同部位组织的检测结果表明, 不仅在含有生殖组织的腹部有这两类共生菌的感染, 在其他非生殖组织中同样也感染了这两类共生菌; 虽然Wolbachia和Cardinium在寄主的各个组织中均有分布, 但是两者在白背飞虱成虫（尤其是雄虫）阶段的动态变化有明显的差异。进一步对Wolbachia宿主特异性WO噬菌体的检测结果表明, 自然种群雄虫中Wolbachia的感染率与不感染个体中WO噬菌体的比率呈明显的负相关。因此推测, 雄虫中Wolbachia感染率相对较低的原因可能是由于Wolbachia基因组中溶原性的WO噬菌体受到某种因素的诱导已转化为裂解性噬菌体。研究结果为进一步揭示Wolbachia和Cardinium双重感染条件下对寄主的生殖调控作用及其机制、 垂直传播规律、 两者之间的相互关系以及进一步的应用研究等方面提供了重要的理论基础。     

华南地区柑橘木虱与柑橘粉虱内共生菌检测及其Wolbachia共生菌的系统发育关系分析

【目的】探明不同地理种群的柑橘木虱Diaphorina citri Kuwayama和柑橘粉虱Dialeurodes citri Ashmead体内昆虫内共生菌的种类及其感染率,并以Wolbachia共生菌为代表,对其系统发育关系进行分析,为今后自共生菌角度研发柑橘木虱和柑橘粉虱的新型防控技术奠定基础。【方法】以16S r DNA、23S r DNA以及wsp为目标基因,利用PCR技术检测采自于广州、湛江、南宁、桂林、厦门的柑橘木虱以及采自广州的柑橘粉虱体内共生菌的种类及其感染率;利用多位点序列分型(MLST)技术和MEGA 5.0软件对不同昆虫样本中的Wolbachia进行系统发育关系分析。【结果】本研究采集的柑橘木虱和柑橘粉虱均含有原生共生菌Portiera和次生共生菌Wolbachia、Cardinium、Rickettsia,但该3种次生共生菌在不同木虱与粉虱种群的感染率有所不同;Arsenophonus只在广州和湛江种群的柑橘木虱中检出。基于wsp基因及MLST基因序列的Wolbachia系统发育分析表明,华南地区柑橘木虱和柑橘粉虱体内的Wolbachia均属于Wolbachia的B大组Con亚组。【结论】不同地理种群的柑橘木虱与柑橘粉虱体内感染的共生菌种类及其感染率不同;Wolabchia共生菌与柑橘木虱寄主不存在协同进化关系,在同一采集点存在Wolbachia通过柑橘寄主在柑橘木虱之间、柑橘木虱与柑橘粉虱之间水平传播的可能性。     

二斑叶螨两种群中Wolbachia诱导的胞质不亲和作用的影响因子比较研究

Wolbachia诱导胞质不亲和（cytoplasmic incompatibility, CI）是对寄主的生殖调控中最常见的一种方式,在不同种群中CI表达的差异较大。以二斑叶螨Tetranychus urticae辽宁兴城（LN）和江苏徐州（JS）两个地理种群为实验材料,经筛选获得100%感染Wolbachia和不感染Wolbachia的品系,通过杂交实验和实时定量PCR的方法研究寄主遗传背景、雄螨日龄、温度以及雄螨体内Wolbachia菌量等因子对我国二斑叶螨体内Wolbachia诱导CI能力的影响。结果表明：1,3,5和7日龄的雄螨诱导的CI的强度没有差异,表明雄螨日龄对我国二斑叶螨体内Wolbachia诱导CI的能力没有影响。二斑叶螨分别放在20℃的低温、 25℃的适温和30℃的高温条件下饲养时,Wolbachia诱导CI的能力也没有任何变化,表明温度对我国二斑叶螨体内Wolbachia诱导CI的能力也没有影响。江苏徐州种群所感染Wolbachia菌量显著高于辽宁兴城种群,并且这两个种群感染Wolbachia菌量都随着雄螨日龄的增大而显著增加,表明Wolbachia菌量对我国二斑叶螨体内Wolbachia诱导CI的能力没有影响;江苏徐州种群内Wolbachia不能诱导CI可能是Wolbachia株系与寄主的遗传背景共同作用的结果。研究结果为进一步了解Wolbachia的生殖调控机理提供了重要依据。     

Symbiont diversity and non-random hybridization among indigenous (Ms) and invasive (B) biotypes of Bemisia tabaci

The whitefly Bemisia tabaci (Hemiptera: Aleyrodidae) is a worldwide pest and a vector of numerous plant viruses. B. tabaci is composed of dozens of morphologically indistinguishable biotypes and its taxonomic status is still controversial. This phloem-feeder harbours the primary symbiont Portiera aleyrodidarum and potentially six secondary symbionts: Cardinium, Arsenophonus, Hamiltonella, Rickettsia, Wolbachia and Fritschea. In the southwest Indian Ocean, La Réunion hosts two biotypes of this species: B (invasive) and Ms (indigenous). A multiplex PCR was developed to study the symbiont community of B. tabaci on La Réunion. Symbiont community prevalence and composition, host mitochondrial and nuclear genetic diversity, as well as host plant and localization, were described on field populations of La Réunion for B and Ms B. tabaci biotypes and their hybrids. A clear association between symbiotypes and biotypes was shown. Cardinium, Arsenophonus and Rickettsia were found in the Ms biotype (73.6%, 64.2% and 3.3%, respectively). Hamiltonella (exclusively) and Rickettsia were found in the B biotype (78% and 91.2%, respectively). Hybrids harboured all symbiotypes found in Ms and B populations, but with a higher prevalence of Ms symbiotypes than expected under random hybridization. An unexpected majority was Cardinium mono-infected (65.6%), and a striking minority (9%) harboured Cardinium/Arsenophonus. In the hybrids only, genetic diversity was linked to symbiotype. Among the hybrids, significant links were found between symbiotypes and: (i) mitochondrial COI sequences, i.e. maternal origin; and (ii) alleles of nuclear microsatellite loci, specific to either Ms or B parental biotype. Taken together, our results suggest that Cardinium and/or Arsenophonus may manipulate the reproduction of indigenous (Ms) with invasive (B) biotypes of Bemisia tabaci.