Diversity of bacteria associated with Hormaphidinae aphids (Hemiptera: Aphididae)

Bacteria are ubiquitous inhabitants of animals.Hormaphidinae is a particular aphid group exhibiting very diverse life history traits.However,the microbiota in this group is poorly known.In the present study,using high-throughput sequencing of bacterial 16S ribosomal RNA gene amplicons,we surveyed the bacterial flora in hormaphidine aphids and explored whether the aphid tribe,host plant and geographical distribution are associated with the distribution of secondary symbionts.The most dominant bacteria detected in hormaphidine species are heritable symbionts.As expected,the primary endosymbiont Buchnera aphidicola is the most abundant symbiont across all species and has cospeciated with its host aphids.Six secondary symbionts were detected in Hormaphidinae.Arsenophonus is widespread in Hormaphidinae species,suggesting the possibility of ancient acquisition of this symbiont.Ordination analyses and statistical tests show that the symbiont composition does not seem to relate to any of the aphid tribes,host plants or geographical distributions,which indicate that horizontal transfers might occur for these symbionts in Hormaphidinae.Correlation analysis exhibits negative interference between Buchnera and coexisting secondary symbionts,while the interactions between different secondary symbionts are complicated.These findings display a comprehensive picture of the microbiota in Hormaphidinae and may be helpful in understanding the symbiont diversity within a group of aphids.     

Pan trapping soybean aphids (Hemiptera: Aphididae) using attractants

Since its introduction into the United States in the past 10 yr, soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), has been a damaging pest to soybean, Glycine max (L.) Merrill. During 2008 and 2009, fields in central and north central Iowa experienced pockets of high soybean aphid populations. Electroantennograms have shown that soybean aphid alatae are capable of detecting host plant volatiles and sex pheromones. Here, we evaluated baited pan traps as a potential soybean aphid attractant. Yellow pan traps were placed in soybean fields after planting along with lures that contained plant volatiles and sex pheromones in 2008 or sex pheromones only in 2009. Pan trap contents were collected weekly, and plant counts also were conducted. Aphids were identified, and soybean aphids were counted to determine whether one chemical lure was more attractive to spring migrants than other lures. In both years, soybean aphids collected in pan traps with lures were not significantly different from the other products tested.     

Within-plant distribution of cotton aphids, Aphis gossypii Glover (Hemiptera: Aphididae), in Bt and non-Bt cotton fields

Knowledge of the vertical and horizontal distribution of Aphis gossypii Glover (Hemiptera: Aphididae) on genetically modified cotton plants over time could help optimize decision-making in integrated cotton aphid management programs. Therefore, the aim of the present study was to determine the vertical and horizontal distribution of A. gossypii in non-transgenic Bt cotton and transgenic Bt-cotton over time during two cotton seasons by examining plants throughout the seasons. There was no significant interaction between years and cotton cultivar treatments for apterous or alate aphids. Considering year-to-year data, analyses on season-long averages of apterous or alate aphids showed that aphid densities per plant did not differ among years. The number of apterous aphids found per plant for the Bt transgenic cultivar (2427 apterous aphids per plant) was lower than for its isoline (3335 apterous aphids per plant). The number of alate aphids found per plant on the Bt transgenic cultivar (12.28 alate aphids per plant) was lower than for the isoline (140.56 alate aphids per plant). With regard to the vertical distribution of apterous aphids or alate aphids, there were interactions between cotton cultivar, plant age and plant region. We conclude that in comparison to non-Bt cotton (DP 4049), Bt cotton (DP 404 BG (Bollgard)) has significant effects on the vertical, horizontal, spatial and temporal distribution patterns of A. gossypii, showing changes in its distribution behaviour inside the plant as the cotton crop develops. The results of our study are relevant for understanding the vertical and horizontal distribution of A. gossypii on Bt cotton cultivar (DP 404 BG (Bollgard)) and on its isoline (DP 4049), and could be useful in decision-making, implementing controls and determining the timing of population peaks of this insect.     

Forecasting migration of cereal aphids (Hemiptera: Aphididae) in autumn and spring

The migration of cereal aphids and the time of their arrival on winter cereal crops in autumn and spring are of particular importance for plant disease (e.g. barley yellow dwarf virus infection) and related yield losses. In order to identify days with migration potentials in autumn and spring, suction trap data from 29 and 45 case studies (locations and years), respectively, were set‐off against meteorological parameters, focusing on the early immigration periods in autumn (22 September to 1 November) and spring (1 May to 9 June). The number of cereal aphids caught in a suction trap increased with increasing temperature, global radiation and duration of sunshine and decreased with increasing precipitation, relative humidity and wind speed. According to linear regression analyses, the temperature, global radiation and wind speed were most frequently and significantly associated with migration, suggesting that they have a major impact on flight activity. For subsequent model development, suction trap catches from different case studies were pooled and binarily classified as days with or without migration as defined by a certain number of migrating cereal aphids. Linear discriminant analyses of several predictor variables (assessed during light hours of a given day) were then performed based on the binary response variables. Three models were used to predict days with suction trap catches ≥1, ≥4 or ≥10 migrating cereal aphids in autumn. Due to the predominance of Rhopalosiphum padi individuals (99.3% of total cereal aphid catch), no distinction between species (R. padi and Sitobion avenae) was made in autumn. As the suction trap catches were lower and species dominance changed in spring, three further models were developed for analysis of all cereal aphid species, R. padi only, and Metopolophium dirhodum and S. avenae combined in spring. The empirical, cross‐classification and receiver operating characteristic analyses performed for model validation showed different levels of prediction accuracy. Additional datasets selected at random before model construction and parameterization showed that predictions by the six migration models were 33–81% correct. The models are useful for determining when to start field evaluations. Furthermore, they provide information on the size of the migrating aphid population and, thus, on the importance of immigration for early aphid population development in cereal crops in a given season.     

Species identification of aphids (Insecta: Hemiptera: Aphididae) through DNA barcodes

A 658-bp fragment of mitochondrial DNA from the 5' region of the mitochondrial cytochrome c oxidase 1 (COI) gene has been adopted as the standard DNA barcode region for animal life. In this study, we test its effectiveness in the discrimination of over 300 species of aphids from more than 130 genera. Most (96%) species were well differentiated, and sequence variation within species was low, averaging just 0.2%. Despite the complex life cycles and parthenogenetic reproduction of aphids, DNA barcodes are an effective tool for identification.     

毛蚜属Chaitophorus蚜虫(半翅目:蚜科:毛蚜亚科)与初级内共生菌Buchnera的演化关系

【目的】蚜虫体内共生菌种类丰富,二者关系十分密切。几乎所有蚜虫都具有一类专性的初级内共生菌Buchnera aphidicola,二者的专性共生关系使蚜虫-Buchnera成为研究共生关系演化的理想模型。本研究对蚜虫-Buchnera在低级阶元水平上的"平行演化假说"进行了验证。【方法】选取在杨属Populus或柳属Salix植物上营同寄主全周期生活的毛蚜属Chaitophorus蚜虫作为研究对象,基于不同来源的分子标记(蚜虫线粒体基因、核基因和内共生菌基因),运用最大似然法和贝叶斯法重建蚜虫和Buchnera的系统树,并利用Tree Map、Jane和Para Fit检验二者是否具有协同系统发生关系。【结果】Tree Map和Jane分析检测到毛蚜属蚜虫与Buchnera具有显著的共成种信号,Para Fit分析结果表明二者的总体关联极为显著。【结论】毛蚜属蚜虫与其初级内共生菌Buchnera在种级及以下水平上符合"平行演化假说",并且二者的演化关系不会受到寄主植物差异的影响。     

Virulence of Hypocreales fungi to pecan aphids (Hemiptera: Aphididae) in the laboratory

There is need for efficacious biocontrol agents for aphids in commercial orchards. As a preliminary step to this end we determined the virulence of several Hypocreales fungi to pecan aphids. In the first experiment we tested the virulence of Isaria fumosorosea (ARSEF 3581) blastospores to three pecan aphids Monellia caryella, Melanocallis caryaefoliae, and Monelliopsis pecanis under laboratory conditions. Rates of 1 × 10⁷ or 1 × 10⁸ spores per ml were applied in 2 ml via a spray tower to 90 mm Petri dishes containing 10 aphids each. Mortality and mycosis were determined after 24, 48 and 72 h. Treatment effects were observed by 48 h post-application, and by 72 h the higher application rate caused >90% mortality and mycosis in M. caryella and M. caryaefoliae, whereas <70% was observed in M. pecanis.We conducted two subsequent experiments (Experiments 2 and 3), using the same methodology, to compare the virulence of several Hypocreales species and strains against the aphid of primary economic concern to most pecan growers, M. caryaefoliae. In Experiment 2, we compared blastospores and conidia of two I. fumosorosea strains (ARSEF 3581 and ATCC 20874 [= strain 97]). The blastospores of ARSEF 3581 and conidia of ATCC 20874 showed higher virulence than other treatments and thus were included in Experiment 3, which also compared the virulence of conidia of Beauveria bassiana (GHA strain) and Metarhizium anisopliae (F52 strain). Results in Experiment 3 indicated the highest virulence in I. fumosorosea 3581 blastospores and M. anisopliae (F52) followed by I. fumosorosea (20874) conidia. The detection of pathogenicity to pecan aphids establishes the potential for commercial usage and additional study. Results reported here will narrow treatments to test in future greenhouse and field trials.     

Rise and fall of cotton aphid (Hemiptera: Aphididae) populations in southeastern cotton production systems

The impact of natural enemies on cotton aphid, Aphis gossypii Glover (Hemiptera: Aphididae), populations in cotton, Gossypium hirsutum L., production systems in the southeastern United States was evaluated over 3 yr in irrigated commercial cotton fields. Fungal epizootics caused by the entomopathogen Neozygites fresenii (Nowakowski) Batko reduced aphid numbers to subthreshold levels in 1999, 2000, and 2001 and occurred consistently in early to mid-July in all 3 yr. Scymnus spp. were the most abundant aphidophagous predators, although other coccinellid species and generalist predators such as spiders, fire ants, heteropterans, and neuropterans also were present. Studies using arthropod exclusion cages demonstrated little impact of predators or parasitoids on aphid populations before fungal epizootics. Arthropod natural enemies were most abundant after epizootics and may have suppressed aphid populations late in the season. Seed cotton yield, and lint quality were not affected by aphicide applications in any year of the study. Implications of these findings for aphid management in the southeastern United States are discussed.     

Development of setal patterns in embryonic and adult aphids of the Hormaphidinae （Hemiptera： Aphididae）

The elementary traits and diversity of embryonic setal patterns of 31 species of 22 genera, in three tribes of Hormaphidinae are studied. The embryonic setal patterns to every species are provided. Based on the parsimony principle and the developmental principle, these patterns are analyzed in each tribe following the terminology of Richards （1965）. It is suggested that the setal patterns of abdominal tergites are the most important characters in embryonic and adult setal patterns. In conclusion, neoteny is a phylogenetic trend in the Hormaphidinae. Nipponaphidini and Hormaphidini have more typical neoteny in each genus and more diversity in different types of adult viviparous females than Cerataphidini, by showing more frequent and various protopatterns or mesopatterns.     

Sources of variation in the interaction between three cereal aphids (Hemiptera: Aphididae) and wheat (Poaceae)

The relative contributions of host plant, herbivore species and clone to variation in the interaction between cereal aphids and wheat were investigated using five clones each of three species, Rhopalosiphum padi (Linnaeus), Sitobion avenae (Fabricius) and Schizaphis graminum (Rondani), on seedlings of two cultivars of Triticum aestivum L. and one cultivar of Triticum durum Desf. More individuals and biomass of R. padi than of the other two species were produced on seedlings. The three wheat cultivars lost similar amounts of biomass as a result of infestation by aphids, with the amount lost depending on aphid species: S. avenae caused the lowest loss in biomass. Variation in aphid biomass production was due mostly to differences among aphid species (70%), less to the interaction between wheat type and aphid species (7%), and least to aphid clone (1%). The specific impact of the aphids on the plants ranged from 1.7 to 3.7 mg of plant biomass lost per mg of aphid biomass gained, being lowest for R. padi and highest for S. graminum. Variation in plant biomass lost to herbivory was due mostly to unknown sources of error (95%), probably phenotypic differences among individual seedlings, with 3% due to aphid species and none attributable to aphid clone. For these aphid-plant interactions, differences among aphid clones within species contributed little to variation in aphid and plant productivity; therefore, a small sample of clones was adequate for quantifying the interactions. Furthermore, one clone of each species maintained in the laboratory for about 200 parthenogenetic generations was indistinguishable from clones collected recently from the field.     

Mitochondrial genome sequences effectively reveal deep branching events in aphids (Insecta: Hemiptera: Aphididae)

Over the past dozen years, considerable effort has been devoted to unravelling the higher‐level phylogenetic relationships of viviparous aphids (Aphididae). However, to date, there is still no commonly accepted phylogenetic hypothesis. In this study, we establish a robust phylogenetic framework for the Aphididae based on mitochondrial genome sequences of 35 aphid species, 22 of which are newly reported. Phylogenetic inferences are performed using multiple data sets, alternative partitioning schemes and different model‐based methods. Our analyses result in well‐supported backbone relationships for the major lineages of aphids, suggesting the feasibility of mitogenome data for resolving phylogenetic questions in aphids. Mindarinae is strongly supported as the earliest branching lineage within Aphididae. A monophyletic clade comprising Calaphidinae, Phyllaphidinae and Saltusaphidinae is corroborated to be the sister group to the species‐richest subfamily Aphidinae. In addition, the morphologically defined subfamily Eriosomatinae is uncovered to be non‐monophyletic.     

Microbiota associated with Mollitrichosiphum aphids (Hemiptera: Aphididae: Greenideinae): diversity,host species specificity and phylosymbiosis

Symbiotic association is universal in nature, and an array of symbionts play a crucial part in host life history. Aphids and their diverse symbionts have become a good model system to study insect-symbiont interactions. Previous symbiotic diversity surveys have mainly focused on a few aphid clades, and the relative importance of different factors regulating microbial community structure is not well understood. In this study, we collected 65 colonies representing eight species of the aphid genus Mollitrichosiphum from different regions and plants in southern China and Nepal and characterized their microbial compositions using Illumina sequencing of the V3 − V4 hypervariable region of the 16S rRNA gene. We evaluated how microbiota varied across aphid species, geography and host plants and the correlation between microbial community structure and host aphid phylogeny. Heritable symbionts dominated the microbiota associated with Mollitrichosiphum, and multiple infections of secondary symbionts were prevalent. Ordination analyses and statistical tests highlighted the contribution of aphid species in shaping the structures of bacterial, symbiont and secondary symbiont communities. Moreover, we observed a significant correlation between Mollitrichosiphum aphid phylogeny and microbial community composition, providing evidence for a pattern of phylosymbiosis between natural aphid populations and their microbial associates.     

Natural occurrence of entomophthoroid fungi (Entomophthoromycota) of aphids (Hemiptera: Aphididae) on cereal crops in Argentina

The spectrum of entomophthoroid fungal species parasitising aphids on cereal crops and a study of the phenology and prevalence of these pathogens were investigated in Argentina. The studies were conducted at six different sites cultivated with crops of Triticum aestivum (wheat), Avena sativa (oats) and Sorghum bicolor (sorghum) during two consecutive years. Entomopathogenic fungi from the new phylum Entomophthoromycota were recorded from six aphid species on cereals in Argentina: Rhopalosiphum maidis, Rhopalosiphum padi, Rhopalosiphum rufiabdominalis, Schizaphis graminum, Sitobion avenae and Sipha maydis. Three species of entomophthoroid fungi were found infecting these aphid species: Pandora neoaphidis, Zoophthora radicans (Entomophthorales: Entomophthoraceae) and Neozygites fresenii (Neozygitales: Neozygitaceae). Entomophthoroid fungal infections occurred mostly in autumn–winter seasons (March–August), and coincided with periods of high relative humidity and comparatively low temperatures. This study represents the first base‐line characterisation of entomophthoroid fungi infecting aphids on cereal crops in Argentina.     

Barcoding aphids (Hemiptera: Aphididae) of the Korean Peninsula: updating the global data set

DNA barcode (mitochondrial COI) sequences are provided for species identification of aphids from the Korean Peninsula. Most (98%) of the 154 species had distinct COI sequences (average 0.05% intraspecific pairwise divergence) relative to the degree of sequence divergence among species (average value 5.84%). For species in common with other regions, barcodes for Korean samples fell near or within known levels of variation. Based on these results, we conclude that DNA barcodes can provide an effective tool for identifying aphid species in such applications as pest management, monitoring and plant quarantine.     

Pathogenicity of hyphomycet fungi to aphids Aphis gossypii Glover and Myzus persicae (Sulzer) (Hemiptera: Aphididae)

The aphids Aphis gossypii and Myzus persicae are cosmopolitan, poliphagous and damage cultivated plants. The effects of the entomopathogenic fungi Beauveria bassiana (isolate IBCB 66), Metarhizium anisopliae (isolate IBCB 121), Paecilomyces fumosoroseus (isolate IBCB 141) and Lecanicillium (= Verticillium) lecanii (isolate JAB 02) on third instar nymphs of A. gossypii and M. persicae were evaluated in the laboratory at 25 degrees C, 70 +/- 10% RH and 12h photophase. The aphids were transferred to petri dishes with a foliar disk (cotton or pepper) with a layer of 1 cm tick of agar-water. The fungi were applied in a suspension containing 1.0 x 106 to 1.0 x 108 conidia/ml. In the control treatment 1 ml of sterilized water was added to the foliar disks. The mortality of aphids was evaluated daily. B. bassiana and M. anisopliae caused 100% mortality at the seventh day after inoculation, for both species. L. lecanii was the fungus that provided mortality later in the aphids and M. persicae was more susceptible to both fungi than A. gossypii.     

Oxydemeton-methyl resistance, mechanisms, and associated fitness cost in green peach aphids (Hemiptera: Aphididae)

Susceptibility to oxydemeton-methyl and imidacloprid, and the inhibitory effects of oxydemeton-methyl and some organophosphate compounds on acetylcholinesterase (AChE) and carboxylesterase activity were studied in two populations (Karaj and Rasht) of green peach aphids, Myzus persicae (Sulzer). Results show that the Karaj population was resistant to oxydemeton-methyl but susceptible to imidacloprid. The esterase activity of the resistant and susceptible populations suggests that one of the resistance mechanisms to oxydemeton-methyl was esterase-based. The inhibition assay shows that the AChE of the Karaj population is less sensitive to oxydemeton-methyl and paraoxon derivatives. Regarding the paraoxon derivatives, the smaller paraoxon side chain is more potent against the modified AChE than against the AChE from the susceptible strain. Fertility life table parameters of green peach aphid populations resistant and susceptible to oxydemeton-methyl also were studied under laboratory conditions. The standard errors of the population growth parameters were calculated using the Jackknife method. Results showed that susceptible strain exhibits a significantly higher r(m) than the resistant strain, probably because the resistant strain had a higher generation time than the susceptible strain. These results suggested that the resistant Karaj strain may be less fit than the susceptible strain.     

Entomopathogenic fungi stimulate transgenerational wing induction in pea aphids,Acyrthosiphon pisum (Hemiptera: Aphididae)

1. Aphid natural enemies include not only predators and parasitoids but also pathogens, of which fungi are the most studied for biological control. While wing formation in aphids is induced by abiotic conditions, it is also affected by biotic interactions with their arthropod natural enemies. Wing induction via interactions with arthropod natural enemies is mediated by the increase in their physical contact when alarmed (pseudo‐crowding). Pathogenic fungi do not trigger this alarm behaviour in aphids and, therefore, no pseudo‐crowding occurs. 2. We hypothesise that, while pathogenic fungi will stimulate maternally induced wing formation, the mechanism is different and is influenced by pathogen specificity. We tested this hypothesis using two entomopathogenic fungi, Pandora neoaphidis and Beauveria bassiana, an aphid specialist and a generalist respectively, on the pea aphid, Acyrthosiphon pisum Harris. 3. We first demonstrate that pea aphids infected with either pathogen and maintained in groups on broad bean plants produced a higher proportion of winged morphs than uninfected control aphids. We then show that, when maintained in isolation, aphids infected with either pathogen also produced higher proportions of winged offspring than control aphids. There was no difference between P. neoaphidis and B. bassiana in their effects on wing induction in either experiment. 4. Unlike the effect of predators and parasitoids on pea aphid wing induction, the effect of pathogens is independent of physical contact with other aphids, suggesting that physiological cues induce wing formation in infected aphids. It is possible that aphids benefit from wing induction by escaping infected patches whilst pathogens may benefit through dispersion. Possible mechanisms of wing induction are discussed.     

Methyl salicylate attracts natural enemies and reduces populations of soybean aphids (Hemiptera: Aphididae) in soybean agroecosystems

Methyl salicylate, an herbivore-induced plant volatile, has been shown to attract natural enemies and affect herbivore behavior. In this study, methyl salicylate was examined for its attractiveness to natural enemies of the soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), and for its direct effects on soybean aphid population growth rates. Methyl salicylate lures were deployed in plots within organic soybean [Glycine max (L.) Merr.] fields. Sticky card traps adjacent to and 1.5 m from the lure measured the relative abundance of natural enemies, and soybean aphid populations were monitored within treated and untreated plots. In addition, exclusion cage studies were conducted to determine methyl salicylate's effect on soybean aphid population growth rates in the absence of natural enemies. Significantly greater numbers of syrphid flies (Diptera: Syrphidae) and green lacewings (Neuroptera: Chrysopidae) were caught on traps adjacent to the methyl salicylate lure, but no differences in abundance were found at traps 1.5 m from the lure. Furthermore, abundance of soybean aphids was significantly lower in methyl salicylate-treated plots. In exclusion cage studies, soybean aphid numbers were significantly reduced on treated soybean plants when all plants were open to natural enemies. When plants were caged, however, soybean aphid numbers and population growth rates did not differ between treated and untreated plants suggesting no effect of methyl salicylate on soybean aphid reproduction and implicating the role of natural enemies in depressing aphid populations. Although aphid populations were reduced locally around methyl salicylate lures, larger scale studies are needed to assess the technology at the whole-field scale.     

Oviposition response of green lacewings (Neuroptera: Chrysopidae) to aphids (Hemiptera: Aphididae) and potential attractants on pecan

Pecan foliage is attacked by three species of aphids [Monellia caryella (Fitch), Melanocallis caryaefoliae (Davis), and Monelliopsis pecanis Bissell], resulting in damage that can reduce tree nut yield. In this study, we assayed the ovipositional response of the green lacewing Chrysoperla rufilabris (Burmeister) to M. caryella and M. caryaefoliae at high and low aphid densities and the development of C. rufilabris larvae when fed solely on each of the three pecan aphid species. During 2004 and 2005, combinations of attractants and food sprays were applied to pecan trees in an orchard to monitor green lacewing ovipositional response. We found that C. rufilabris laid more eggs on seedling trees infested with the M. caryella (at both high and low densities) than on seedlings infested with M. caryaefoliae. Development of C. rufilabris was unaffected by aphid species. At least one attractant/food spray treatment applied to trees in an orchard significantly increased green lacewing oviposition for three of the five treatment dates over both years. These results show that larvae of C. rufilabris will consume all aphid species attacking pecan, even though female ovipositional response can differ for aphid species. It is likely that combinations of attractants and food sprays can be used to enhance green lacewing populations in orchards.