Search Results (13)

The cotton aphid, Aphis gossypii, is a globally significant agricultural pest whose microbiota plays vital roles in its physiology and adaptation. However, the dynamics of bacterial communities across its developmental stages remain poorly understood. This study employed full-length 16S rRNA gene sequencing to characterize the microbiota structure, diversity, and functional potential in nine developmental stages of A. gossypii, including egg, nymph (1-, 3-, 5-, 7-day-old), and adult (1-, 3-, 5-, 7-day-old). Results revealed Proteobacteria (72.75–95.51%) as the dominant phylum across all stages, with Buchnera aphidicola (primary obligate symbiont) constituting over 23.83% of bacterial abundance and peaking in eggs (≈80%). Alpha diversity indices (Shannon, Simpson) indicated significantly higher microbial diversity in nymphs compared to adults, suggesting stage-specific ecological interactions. While beta diversity analysis showed no structural clustering by developmental stage, functional predictions highlighted enrichment in metabolic pathways (>73% of genes), though limitations in 16S-based functional inference were noted. Notably, facultative symbionts like Hamiltonella or Serratia were absent, contrasting with other aphid systems. Dynamic shifts in Buchnera titer and the prominence of Delftia tsuruhatensis and Enterobacter hormaechei implied potential roles in host adaptation. These findings highlight the persistent dominance of the obligate symbiont Buchnera aphidicola across all developmental stages, despite quantitative fluctuations in its abundance, alongside stage-specific shifts in facultative bacterial communities, offering insights into novel targets for microbiome-driven pest management strategies. Further multi-omics approaches are warranted to validate functional contributions of these microbial communities. Full article

The greenbug aphid (Schizaphis graminum (Rondani)) is a major pest of wheat and an important vector of wheat viruses. An RNA-seq study was conducted to investigate the microbial effects of two greenbug genotypes, the presence or absence of cereal yellow dwarf virus, and the condition of the wheat host over a 20-day time course of unrestricted greenbug feeding. Messenger RNA reads were mapped to ca. 47,000 bacterial, 1218 archaeal, 14,165 viral, 571 fungal, and 94 protozoan reference or representative genomes, plus greenbug itself and its wheat host. Taxon counts were analyzed with QIIME2 and DESeq2. Distinct early (days 1 through 10) and late (days 15 and 20) communities differed in the abundance of typical enteric genera (Shigella, Escherichia, Citrobacter), which declined in the late community, while the ratio of microbial to greenbug read counts declined 50% and diversity measures increased. The nearly universal aphid endosymbiont, Buchnera aphidicola, accounted for less than 25% of the read counts in both communities. There were 302 differentially expressed (populated) genera with respect to early and late dates, while 25 genera differed between the greenbug genotypes and nine differed between carrier and virus-free greenbugs. The late community was likely responding to starvation as the wheat host succumbed to aphid feeding. Our results add to basic knowledge about aphid microbiomes and offer an attractive alternative method to assess insect microbiomes. Full article

Buchnera aphidicola, an obligate endosymbiont of most aphid species, can influence aphids’ host adaptability through amino acid metabolism, potentially mediating biotype differentiation. However, its role in the biotype differentiation of Sitobion avenae remains unclear. To address this issue, six S. avenae biotypes were tested in this study. Buchnera abundance varied among biotypes fed on different wheat/barley varieties (i.e., Zhong 4 wumang, 186-TM12-34; Dulihuang, Zaoshu No.3, Xiyin No.2). The reduction in Buchnera abundance through antibiotic (rifampicin) treatment altered the virulence of five S. avenae biotypes. Based on transcriptome analysis, the differential expression of three genes (i.e., LeuB, TrpE, and IlvD) related to leucine, tryptophan, isoleucine, and valine metabolism was detected between different biotypes. Principal component analysis showed that leucine and tryptophan deficiencies most significantly impacted nymph development duration and aphid fecundity. Additionally, a neighbor-joining phylogenetic tree indicated the genetic differentiation of Buchnera among different biotypes. These results suggest Buchnera-mediated amino acid metabolism is correlated with biotype differentiation in S. avenae, although the precise mechanisms by which Buchnera influences this differentiation require further investigation. This study can offer a theoretical basis for the development of resistant crops, leading to the sustainable control of this aphid and reduced reliance on chemical insecticides. Full article

Gut microbiomes profoundly influence insect health and mediate interactions between plant hosts and their environments. Insects, including aphids, harbour diverse obligate symbionts that synthesize essential nutrients and facultative symbionts that enhance host fitness in specific ecological contexts. Sorghum (Sorghum bicolor) is a significant cereal crop cultivated worldwide that has been negatively affected by the presence of an invasive piercing-sucking insect pest, the sugarcane aphid (SCA; Melanaphis sacchari). We previously identified SC265 and SC1345 as the resistant and susceptible sorghum lines, respectively, among the founder nested association mapping (NAM) population. Here, using these resistant and susceptible lines, we explored variations in the SCA gut microbiome when they feed on two different sorghum lines with varied resistance levels. Analyses after excluding the obligate endosymbiont Buchnera aphidicola from the dataset showed a significant difference in microbial diversity and composition between resistant and susceptible sorghum lines 7- and 14 days post aphid infestation. Our results indicate that the SCA fed on susceptible and resistant sorghum lines had Pseudomonadaceae and Rhizobiaceae, respectively, as the most abundant bacterial families. Differences in gut microbial community composition were underscored by alpha diversity metrics and beta diversity compositional analyses. These findings contribute to our understanding of the intricate interplay between plant and aphid microbiomes, shedding light on potential avenues to bolster sorghum resistance to SCA. Full article

The barley yellow dwarf virus (BYDV) of cereals is thought to substantially increase the high-temperature tolerance of its aphid vector, Rhopalosiphum padi, which may enhance its transmission efficiency. This is based on experiments with North American strains of BYDV and R. padi. Here, we independently test these by measuring the temperature tolerance, via Critical Thermal Maximum (CTmax) and knockdown time, of Australian R. padi infected with a local BYDV isolate. We further consider the interaction between BYDV transmission, the primary endosymbiont of R. padi (Buchnera aphidicola), and a transinfected secondary endosymbiont (Rickettsiella viridis) which reduces the thermotolerance of other aphid species. We failed to find an increase in tolerance to high temperatures in BYDV-infected aphids or an impact of Rickettsiella on thermotolerance. However, BYDV interacted with R. padi endosymbionts in unexpected ways, suppressing the density of Buchnera and Rickettsiella. BYDV density was also fourfold higher in Rickettsiella-infected aphids. Our findings indicate that BYDV does not necessarily increase the temperature tolerance of the aphid transmission vector to increase its transmission potential, at least for the genotype combinations tested here. The interactions between BYDV and Rickettsiella suggest new ways in which aphid endosymbionts may influence how BYDV spreads, which needs further testing in a field context. Full article

The green peach aphid, Myzus persicae, is globally distributed and an important pest of many economically valuable food crops, largely due to its ability to transmit plant viruses. Almost all aphids, including M. persicae, carry the obligate symbiont Buchnera aphidicola, which provides essential amino acids that aphids cannot obtain from the phloem of plants themselves. Many aphids also harbor facultative (secondary) endosymbionts, which provide benefits under specific ecological conditions. In this study, we screened for secondary endosymbionts in M. persicae, with a particular focus on Australian populations where this species is growing in status as a major agricultural pest. We compared 37 Australian M. persicae populations with other populations, including 21 field populations from China and 15 clones from the UK, France, Italy, Greece, USA, Spain, South Korea, Chile, Japan and Zimbabwe. No secondary endosymbionts were identified in M. persicae samples outside of China, despite samples covering a wide geographic range and being collected from several host plant families. We detected two secondary endosymbionts (Rickettsia, Spiroplasma) in Chinese samples, although diversity appeared lower than detected in a recent study. We also found very high clonal diversity in Chinese samples based on DNA microsatellite markers in comparison with lower clonal diversity from Australia. These patterns may indicate a higher diversity of secondary endosymbionts (and clonal diversity) in the native range of M. persicae when compared to its invasive range. Full article

In this study, Russian olive trees exhibiting witches’-broom symptoms were collected from urban green areas in Tabriz, in the northwest of Iran. PCR analysis confirmed that phytoplasma caused the disease and, according to the resulting Sanger sequencing electropherogram, a mixed infection with two or more phytoplasma species within the Russian olive trees was revealed. Next-generation sequencing analyses, using the Illumina method, were performed on total DNA from the infected Russian olive plants to recognize the microbial genomic content and assemble the whole genome of the causative pathogen(s). The use of MetaphlAn2 and Kraken2 to analyze species composition revealed the very diverse and unique compositions of different Prokaryotic and Eukaryotic species within the infected plants. Several bacteria and fungi were discovered inside the samples, among which Mycoplasmatota was significantly dominating. Interestingly, the results also revealed a high level of endosymbiotic bacteria and Archaea (Methanobacteria) genome contents within the samples. Bowtie2, metaSPAdes, and CD-HIT pipelines were used to perform the initial genome assembly, and the whole genome of the notable phytoplasma species was assembled and submitted to Genbank. Full article

Dependence on multiple nutritional bacterial symbionts forming a metabolic unit has repeatedly evolved in many insect species that feed on nutritionally unbalanced diets such as plant sap. This is the case for aphids of the subfamilies Lachninae and Chaitophorinae, which have evolved di-symbiotic systems in which the ancient obligate nutritional symbiont Buchnera aphidicola is metabolically complemented by an additional nutritional symbiont acquired more recently. Deciphering how different symbionts integrate both metabolically and anatomically in such systems is crucial to understanding how complex nutritional symbiotic systems function and evolve. In this study, we sequenced and analyzed the genomes of the symbionts B. aphidicola and Serratia symbiotica associated with the Chaitophorinae aphids Sipha maydis and Periphyllus lyropictus. Our results show that, in these two species, B. aphidicola and S. symbiotica complement each other metabolically (and their hosts) for the biosynthesis of essential amino acids and vitamins, but with distinct metabolic reactions supported by each symbiont depending on the host species. Furthermore, the S. symbiotica symbiont associated with S. maydis appears to be strictly compartmentalized into the specialized host cells housing symbionts in aphids, the bacteriocytes, whereas the S. symbiotica symbiont associated with P. lyropictus exhibits a highly invasive phenotype, presumably because it is capable of expressing a larger set of virulence factors, including a complete flagellum for bacterial motility. Such contrasting levels of metabolic and anatomical integration for two S. symbiotica symbionts that were recently acquired as nutritional co-obligate partners reflect distinct coevolutionary processes specific to each association. Full article

High-temperature events are evidenced to exert significant influence on the population performance and thermal biology of insects, such as aphids. However, it is not yet clear whether the bacterial symbionts of insects mediate the thermal tolerance traits of their hosts. This study is intended to assess the putative association among the chronic and acute thermal tolerance of two cereal aphid species, Rhopalosiphum padi (L.) and Sitobion avenae (F.), and the abundance of their bacterial symbionts. The clones of aphids were collected randomly from different fields of wheat crops and were maintained under laboratory conditions. Basal and acclimated CTmax and chronic thermal tolerance indices were measured for 5-day-old apterous aphid individuals and the abundance (gene copy numbers) of aphid-specific and total (16S rRNA) bacterial symbionts were determined using real-time RT-qPCR. The results reveal that R. padi individuals were more temperature tolerant under chronic exposure to 31 °C and also exhibited about 1.0 °C higher acclimated and basal CTmax values than those of S. avenae. Moreover, a significantly higher bacterial symbionts’ gene abundance was recorded in temperature-tolerant aphid individuals than the susceptible ones for both aphid species. Although total bacterial (16S rRNA) abundance per aphid was higher in S. avenae than R. padi, the gene abundance of aphid-specific bacterial symbionts was nearly alike for both of the aphid species. Nevertheless, basal and acclimated CTmax values were positively and significantly associated with the gene abundance of total symbiont density, Buchnera aphidicola, Serratia symbiotica, Hamilton defensa, Regiella insecticola and Spiroplasma spp. for R. padi, and with the total symbiont density, total bacteria (16S rRNA) and with all aphid-specific bacterial symbionts (except Spiroplasma spp.) for S. avenae. The overall study results corroborate the potential role of the bacterial symbionts of aphids in conferring thermal tolerance to their hosts. Full article

Pea aphid (Acyrthosiphon pisum) is a worldwide pest that feeds exclusively on the phloem sap of numerous host plants. It harbours a well-known primary endosymbiont Buchnera aphidicola that helps to overcome the nutritional deficiency of a plant-based diet. However, how the Buchnera contributes to the nutritional and energy metabolism of its aphid host is unclear to date. In the current study, the function of Buchnera in relation to nutritional synthesis of pea aphid was investigated by disrupting the primary endosymbiont with an antibiotic rifampicin. Our findings revealed that the disruption of Buchnera led to infertility and higher loss in body mass of aphid hosts. Body length and width were also decreased significantly compared to healthy aphids. The detection of nutrition indicated that the quantity of proteins, soluble sugars, and glycogen in aposymbiotic pea aphids increased slowly with the growth of the aphid host. In comparison, the quantities of all the nutritional factors were significantly lower than those of symbiotic pea aphids, while the quantity of total lipid and neutral fat in aposymbiotic pea aphids were distinctly higher than those of symbiotic ones. Thus, we concluded that the significant reduction of the total amount of proteins, soluble sugars, and glycogen and the significant increase of neutral fats in aposymbiotic pea aphids were due to the disruption of Buchnera, which confirmed that the function of Buchnera is irreplaceable in the pea aphid. Full article

The domino theory of gene loss states that when some particular gene loses its function and cripples a cellular function, selection will relax in all functionally related genes, which may allow for the non-functionalization and loss of these genes. Here we study the role of epistasis in determining the pattern of gene losses in a set of genes participating in cell envelope biogenesis in the endosymbiotic bacteria Buchnera aphidicola. We provide statistical evidence indicating pairs of genes in B. aphidicola showing correlated gene loss tend to have orthologs in Escherichia coli known to have alleviating epistasis. In contrast, pairs of genes in B. aphidicola not showing correlated gene loss tend to have orthologs in E. coli known to have aggravating epistasis. These results suggest that during the process of genome reduction in B. aphidicola by gene loss, positive or alleviating epistasis facilitates correlated gene losses while negative or aggravating epistasis impairs correlated gene losses. We interpret this as evidence that the reduced proteome of B. aphidicola contains less pathway redundancy and more compensatory interactions, mimicking the situation of E. coli when grown under environmental constrains. Full article

It is known that PCR amplification of highly homologous genes from complex DNA mixtures can generate a significant proportion of chimeric sequences. The 16S rRNA gene is not only widely used in estimating the species diversity of endosymbionts in aphids but also used to explore the co-diversification of aphids and their endosymbionts. Thus, chimeric sequences may lead to the discovery of non-existent endosymbiont species and mislead Buchnera-based phylogenetic analysis that lead to false conclusions. In this study, a high probability (6.49%) of chimeric sequence occurrence was found in the amplified 16S rRNA gene sequences of endosymbionts from aphid species in the subfamily Lachninae. These chimeras are hybrid products of multiple parent sequences from the dominant species of endosymbionts in each corresponding host. It is difficult to identify the chimeric sequences of a new or unidentified species due to the high variability of their main parent, Buchnera aphidicola, and because the chimeric sequences can confuse the phylogenetic analysis of 16S rRNA gene sequences. These chimeras present a challenge to Buchnera-based phylogenetic research in aphids. Thus, our study strongly suggests that using appropriate methods to detect chimeric 16S rRNA sequences may avoid some false conclusions in endosymbiont-based aphid research. Full article

Obligate nutritional symbioses require balance between the energetic needs of the host and the symbiont. The resident symbiont population size within a host may have major impacts on host fitness, as both host and symbiont consume and supply metabolites in a shared metabolite pool. Given the massive genome degradation that is a hallmark of bacterial endosymbionts of insects, it is unclear at what level these populations are regulated, and how regulation varies among hosts within natural populations. We measured the titer of the endosymbiont Buchnera aphidicola from different clones of the pea aphid, Acyrthosiphon pisum, and found significant variation in titer, measured as Buchnera genomes per aphid genome, among aphid clones. Additionally, we found that titer can change with the age of the host, and that the number of bacteriocytes within an aphid is one factor likely controlling Buchnera titer. Buchnera titer measurements in clones from a sexual cross indicate that the symbiont genotype is not responsible for variation in titer and that this phenotype is likely non-heritable across sexual reproduction. Symbiont titer is more variable among lab-produced F₁ aphid clones than among field-collected ones, suggesting that intermediate titer is favored in natural populations. Potentially, a low heritability of titer during the sexual phase may generate clones with extreme and maladaptive titers each season. Full article