Occurrence and Transmission of Facultative Endosymbionts in Aphids

The occurrence of a secondary bacterial symbiont (PASS) of pea aphid, Acyrthosiphon pisum (Harris), was detected by polymerase chain reaction (PCR) with specific nucleotide primers based on PASS 16S rDNA nucleotide sequences from over 80% (50/57) of clones of pea aphid collected from widely separated locations in California. PASS was also detected by PCR in both red and green phenotypes of rose aphid, Macrosiphum rosae (L.), but not in six other species of aphids examined, including blue alfalfa aphid (A. kondoi Shinji). The nucleotide sequences of the PCR-amplified, partial 16S rDNAs (1060 bp) from pea aphid and rose aphid were identical and 99.9% similar to the published 16S rDNA of PASS. PASS and a recently described new rickettsia of pea aphid (PAR) were transmitted by needle injection of hemolymph from positive pea aphid clones into negative clones and into blue alfalfa aphids. Both PASS and PAR were maintained in the offspring of some of the injected mother aphids via high rate of maternal transmission. Received: 18 September 1996 / Accepted: 30 September 1996     

Temperature effect on the growth of <Emphasis Type="Italic">Buchnera</Emphasis> endosymbiont in <Emphasis Type="Italic">Aphis craccivora</Emphasis> (Hemiptera: Aphididae)

Effect of temperature on the growth of the primary endosymbiont Buchnera aphidicola in the cowpea aphid Aphis craccivora was studied by measuring quantitatively the copy number of 16S rDNA of this endosymbiont. A 1.5 kb segment of eubacterial 16S rDNA amplified by PCR from total DNA of Aphis craccivora was confirmed by RFLP analysis and sequence BLAST as that of Buchnera aphidicola. No secondary endosymbiont was detected in the aphid population studied. The relative levels of Buchnera ratio, quantified by real-time PCR, were higher in old nymphs than in young ones at temperatures between 10–30˚C, and this age-dependent difference was more pronounced at lower temperatures. Throughout the entire reproductive stage of Aphis craccivora, the relative levels of Buchnera ratio were higher at 10–25˚C than at 30˚C and 35˚C. A close relationship was found between these levels and the net reproductive rate (R ₀ ) of aphid, which was suppressed not only at 35˚C but also at 10˚C. The decoupling of Aphis craccivora and Buchnera response at low temperatures suggests that the cowpea aphid was more sensitive to low temperatures, while Buchnera was more sensitive to high temperatures.     

Genetic variation in the effect of a facultative symbiont on host-plant use by pea aphids

Ecological specialisation on different host plants occurs frequently among phytophagous insects and is normally assumed to have a genetic basis. However, insects often carry microbial symbionts, which may play a role in the evolution of specialisation. The bacterium Regiella insecticola is a facultative symbiont of pea aphids (Acyrthosiphon pisum) where it is found most frequently in aphid clones feeding on Trifolium giving rise to the hypothesis that it may improve aphid performance on this plant. A study in which R. insecticola was eliminated from a single naturally infected aphid clone supported the hypothesis, but a second involving two aphid clones did not find the same effect. We created a series of new pea aphid–R. insecticola associations by injecting different strains of bacteria into five aphid clones uninfected by symbionts. For all aphid clones, the bacteria decreased the rate at which aphids accepted Vicia faba as a food plant and reduced performance on this plant. Their effect on aphids given Trifolium pratense was more complex: R. insecticola negatively affected acceptance by all aphid clones, had no effect on the performance of four aphid clones, but increased performance of a fifth, thus demonstrating genetic variation in the effect of R. insecticola on pea aphid host use. We discuss how these results may explain the distribution and frequency of this symbiont across different aphid populations. Julia Ferrari and Claire L. Scarborough contributed equally to the work.     

High variation in host adaptation among clones of the pea aphid,Acyrthosiphon pisum on peas,Pisum sativum

The performance of one clone of the pea aphid,Acyrthosiphon pisum (Harris), was assessed on 37 different cultivars and species ofPisum L. In addition, random samples of 36 pea aphid clones collected on alfalfa and clover were tested on a selection of fivePisum sativum L. cultivars. Aphid performance was evaluated in terms of the mean relative growth rate (MRGR) during the first five days of life or other life history variables. The MRGR of the first-mentioned pea aphid clone differed little between cultivars. No significant differences in MRGR were found between wild and cultivatedPisum species or between modern and oldP. sativum cultivars. There was considerable variation in host adaptation among the 36 pea aphid clones within each sampled field. The pea aphid clones showed no consistent pattern in performance on four of the five pea cultivars i.e. there was a significant pea aphid genotype —pea genotype interaction. On one of the cultivars all clones performed well. Pea aphid clones collected from red clover generally performed relatively poorly on pea cultivars, in contrast to the pea aphid clones collected on alfalfa. There was no difference in performance between the two pea aphid colour forms tested. Possible reasons for the high variation and the observed adaptation patterns are discussed. The fact that all clones were collected in two adjacent fields indicates thatA. pisum shows high local intraspecific variability in terms of host adaptation.     

Use of 16S rDNA Sequences as Signature Characters to Identify Xylella fastidiosa

The nucleotide sequences of 16S rDNAs (coding for the small subunit ribosomal RNAs) were used to identify Xylella fastidiosa, a nutritionally fastidious plant pathogenic bacterium. The near-complete 16S rDNAs from nine strains of Xyl. fastidiosa, including seven pathotypes and one strain of Xanthomonas campestris pv. campestris, were amplified through PCR with two conserved primers (forward primer 5′-AGA GTT TGA TCC TGG CTC AG-3′ and reverse primer 5′-AAG GAG GTG ATC CAG CC-3′) and sequenced. The 16S sequences were compared with all eukaryote and prokaryote DNA entries in GenBank database. A Xyl. fastidiosa 16S rDNA sequence, M26601, was determined to be the most similar to all the near-complete (1537 bp) and partial 5′ end sequences from Xyl. fastidiosa, but not those from the Xanthomonas strain. A 20-bp oligonucleotide (5′-TTG GTA GTA ATA CCA TGG GT-3′) was found to be highly characteristic of Xyl. fastidiosa. Since the 16S rDNA of Xyl. fastidiosa strains are highly homologous and characteristically different from other bacteria, including the most closely related Xanthomonas, 16S rDNA sequences can be used as signature characters to identify this bacterium. Received: 22 June 1999 / Accepted: 2 August 1999     

Rickettsia Symbiont in the Pea Aphid Acyrthosiphon pisum: Novel Cellular Tropism, Effect on Host Fitness, and Interaction with the Essential Symbiont Buchnera

In natural populations of the pea aphid Acyrthosiphon pisum, a facultative bacterial symbiont of the genus Rickettsia has been detected at considerable infection frequencies worldwide. We investigated the effects of the Rickettsia symbiont on the host aphid and also on the coexisting essential symbiont Buchnera. In situ hybridization revealed that the Rickettsia symbiont was specifically localized in two types of host cells specialized for endosymbiosis: secondary mycetocytes and sheath cells. Electron microscopy identified bacterial rods, about 2 μm long and 0.5 μm thick, in sheath cells of Rickettsia-infected aphids. Virus-like particles were sometimes observed in association with the bacterial cells. By an antibiotic treatment, we generated Rickettsia-infected and Rickettsia-eliminated aphid strains with an identical genetic background. Comparison of these strains revealed that Rickettsia infection negatively affected some components of the host fitness. Quantitative PCR analysis of the bacterial population dynamics identified a remarkable interaction between the coexisting symbionts: Buchnera population was significantly suppressed in the presence of Rickettsia, particularly at the young adult stage, when the aphid most actively reproduces. On the basis of these results, we discussed the possible mechanisms that enable the prevalence of Rickettsia infection in natural host populations in spite of the negative fitness effects observed in the laboratory.     

Characterization of a Facultative Endosymbiotic Bacterium of the Pea Aphid Acyrthosiphon pisum

The pea aphid U-type symbiont (PAUS) was investigated to characterize its microbiological properties. Fluorescence in situ hybridization (FISH) and electron microscopy revealed that PAUS was a rod-shaped bacterium found in three different locations in the body of the pea aphid Acyrthosiphon pisum: sheath cells, secondary mycetocytes, and hemolymph. Artificial transfer experiments revealed that PAUS could establish stable infection and vertical transmission when introduced into uninfected pea aphids. When 28 aphid species collected in Japan were subjected to a diagnostic PCR assay, four species of the subfamily Aphidinae (Aphis citricola, Aphis nerii, Macrosiphum avenae, and Uroleucon giganteus) and a species of the subfamily Pemphiginae (Colopha kansugei) were identified to be PAUS-positive. The sporadic incidences of PAUS infection without reflecting the aphid phylogeny can be best explained by occasional horizontal transfers of the symbiont across aphid lineages.     

Rickettsial Relative Associated with Papaya Bunchy Top Disease

The phylogeny of a previously unidentified, obligate laticifer-inhabiting bacterium associated with the papaya bunchy top disease was investigated. Portions of genes corresponding to those for 16S rRNA, the flavoprotein subunit of succinate dehydrogenase (SdhA), citrate synthase (GltA), and the 17-kDa rickettsial common antigen were isolated and sequenced from the non-cultivable bacterium from diseased plants. Comparative sequence analyses consistently indicated that the bacterium is a member of the α-subdivision of the Proteobacteria and of the genus Rickettsia. The rickettsia was detected by polymerase chain reaction in diseased, but not healthy, papaya tissues and in the leafhopper vector, Empoasca papayae, providing further evidence of the possible etiological role of the bacterium in the disease. Although Rickettsia have been found naturally in arthropods and can be pathogenic to humans and other vertebrates, this is the first evidence of its kind implicating a Rickettsia as a plant pathogen. Received: 21 July 1997 / Accepted: 28 July 1997     

The community composition of root-associated bacteria of the tomato plant

The objective of this study was to characterize the bacterial community composition in the bulk soil, rhizosphere soil and root tissue of the tomato plant (Lycopersicum esculentum Mill). 16S ribosomal DNA (rDNA) from the bacterial community was amplified using PCR, and sequence analysis of 16S rDNA clones was subsequently used for bacterial identification and phylogenetic classification. Phylogenetic analysis of clones (total of 68) from the bulk soil, rhizosphere and root tissues showed that about 50% of the bacteria belonged to the α-, β-, γ-, and δ-Proteobacteria or Cytophaga–Flavobacterium–Bacteroides (CFB) phyla, with only one high G+C clone identified. A number of diverse bacteria were identified within Proteobacteria, while 87% of the bacteria belonged to the genus Flavobacterium within the CFB phylum, which is a unique finding for tomato plants. Our results will be of interest to those wanting to identify bacteria that can promote plant growth or resistance to diseases.     

Identification of phytoplasmas from Neoaliturus haematoceps associated with sesame phyllody disease in southwestern Turkey

Phytoplasmas were detected based on nested PCR of the F2nR2 region of the 16S rDNA from Neoaliturus haematoceps (Mulsant and Rey) (Family: Cicadellidae). A total of 65 insect samples collected from sesame fields in Antalya, Turkey, during 2012–2014 were tested for phytoplasma detection. Phytoplasmas detected in fifteen samples showed an amplicon approximately 1250 bp in size using the universal primers of P1/P7 and R16F2n/R16R2. Identification of the phytoplasmas by sequence analysis revealed three different 16S rDNA phytoplasma groups: the peanut witches’‐broom, group II; clover proliferation, group VI; and pigeon pea witches’‐broom, group IX. The molecular characterization of subgroups was determined by sequence analysis and PCR‐RFLP using the restriction enzymes RsaI and TaqI. Restriction profiles of the subgroups were also confirmed using the iPhyclassifier program. BLAST and PCR‐RFLP analyses classified the subgroups as II‐D, VI‐A and IX‐C. This is the first report of molecular detection of three 16S rDNA subgroups of phytoplasmas, II‐D, VI‐A and IX‐C, from N. haematoceps in Turkey. This study also supports earlier studies of sesame phyllody phytoplasmas by N. haematoceps.     

Molecular diversity of microbial community in acid mine drainages of Yunfu sulfide mine

Two acid mine drainage (AMD) samples were studied by a PCR-based cloning approach, which were from Yunfu sulfide mine in Guangdong province, China. A total of 15 operational taxonomic units (OTUs) were obtained from the two AMD samples. The percentage of overlapped OTUs in two AMD samples was 42.1%. Phylogenetic analysis revealed that the bacterium in the two samples fell into four putative divisions, which were Nitrospira, α-Proteobacteria, β-Proteobacteria, and γ-Proteobacteria four families. Organisms of genuses Acidithiobacillus and Gallionella, which were in γ-Proteobacteria family and β-Proteobacteria family, respectively, were dominant in two samples. The proportions of clones affiliated with Gallionella in each sample were 47.2% (G2) and 16.9% (G1). The result suggested that organisms of Gallionella were a very important composition in microbial communities of the two AMD samples we studied. In addition, the PCR amplification of archaeal 16S rDNA genes form these two AMD samples have been performed with two sets of archaea-specific primers, but no PCR product found. Zhiguo He, Shengmu Xiao, Xuehui Xie, and Hui Zhong equally contributed to this work.     

Diversity of Endosymbionts in the Potato Psyllid, <Emphasis Type="Italic">Bactericera cockerelli</Emphasis> (Hemiptera: Triozidae), Vector of Zebra Chip Disease of Potato

Zebra chip disease is an emerging, serious disease of solanaceous crops and the causal agent is a bacterium “Candidatus Liberibacter solanacearum” (CLs), also known as “Candidatus Liberibacter psyllaurous”, which is transmitted by the potato psyllid, Bactericera cockerelli (Šulc). We performed bacterial tag-encoded FLX amplicon pyrosequencing (bTEFAP) of the 16S rDNA genes to determine the bacterial microbiota in adult insects from CLs-uninfected and CLs-infected strains of B. cockerelli and potato leaf samples. We obtained sequences from five bacterial species among the two psyllid strains, including “Candidatus Carsonella ruddii”, Wolbachia, CLs, and two transient bacteria, Acinetobacter and Methylibium. We did not detect any common bacteria between psyllids and potato leaf samples using pyrosequencing. We performed PCR analysis using species-specific 16S rDNA primers to confirm pyrosequencing results in individual psyllids including eggs, early-instars, late-instars, and adults of both sexes from both CLs-uninfected and CLs-infected psyllid strains. The primary endosymbiont, “Candidatus Carsonella ruddii” and Wolbachia were detected in all life-stages and sexes of both strains using PCR analyses. The percentage of CLs-infected individuals increased from early-instar (0%), late-instar (40%) until adulthood (60%) in the CLs-infected strain. We believe that CLs levels in early-instars are probably too low to be detected by standard PCR. Using PCR analyses, we confirmed the presence of Acinetobacter in CLs-uninfected and CLs-infected adults (75 and 25%, respectively) but not Methylibium. Further, we detected Acinetobacter in potato leaves using PCR indicating that the psyllids may have acquired this bacterium via feeding on the host plant.     

Molecular detection of the entomopathogenic bacterium Pseudomonas entomophila using PCR

Aims: To develop a specific, fast and simple molecular method useful to detect the entomopathogenic bacterium Pseudomonas entomophila. Methods and Results: The use of bioinformatics tools allowed the identification of unique genes present in P. entomophila genome. Using such genes, we designed primers aiming to detect specifically P. entomophila by PCR. Furthermore, a pair of primers specifically designed to amplify the 16S rRNA gene in Pseudomonas species was used. Primer specificity was checked using environmental pseudomonad and nonpseudomonad species. A 618 ‐bp fragment was amplified only in Pseudomonas using the 16S rDNA primers. Primers (PSEEN1497) designed to detect P. entomophila amplified a 570 ‐bp fragment only in P. entomophila. A duplex PCR was developed combining 16S rDNA and PSEEN1497 primers that allowed the detection of P. entomophila present in experimentally infected Drosophila melanogaster. Conclusions: We developed a molecular method useful to detect P. entomophila present in bacterial cultures or directly from infected insects. Significance of the Study: To the best of our knowledge, this is the first molecular method aiming to detect P. entomophila in environmental samples. The use of our method will facilitate studies related to ecology and insect host range of this entomopathogenic bacterium.     

Fitness effects of two facultative endosymbiotic bacteria on the pea aphid,Acyrthosiphon pisum,and the blue alfalfa aphid,A. kondoi

The effects of two bacterial endosymbionts, designated PASS and PAR, were evaluated on the pea aphid, Acyrthosiphon pisum (Harris) (Hemiptera:Aphididae), in which they occur facultatively, and on the blue alfalfa aphid, A. kondoi Shinji, in which these bacteria have not been found in natural populations. Subclones of pea aphids and blue alfalfa aphids, derived from parent aphid clones that did not contain PASS or PAR, were infected with one or both bacteria, generating PASS- and/or PAR-positive subclones with minimal genetic differences from the parent clones. Under laboratory conditions at 20 °C, PAR consistently reduced the fecundity (by between 19 and 60%) of subclones derived from three different parent pea aphid clones on bur clover, Medicago hispida Gaertn. PAR had intermediate effects on pea aphids reared on sweet pea, Lathyrus odoratus L., and had no significant effect on pea aphids on alfalfa, Medicago sativa L. The effect of PASS was either neutral or negative, depending on parent clone as well as host plant. Also at 20 °C, PASS reduced fecundity (70–77%) and longevity (40–48%), and increased the age of first reproduction (by up to 1.5 days) of blue alfalfa aphid reared on alfalfa and clover. PAR had a less dramatic effect (e.g., 30–39% reduction in fecundity) on these traits of blue alfalfa aphid. In contrast, PAR and PASS increased the fitness of pea aphid subclones of one parent clone reared for three generations at 25 °C on each of the three test plants. Without facultative bacteria, fecundity of the parent clone was reduced to a mean total of < 6 offspring per adult at this elevated temperature, but with PASS or PAR, mean total fecundity of its subclones was > 35. However, this ameliorative effect of facultative bacteria at 25 °C was not found for two other sets of parent clones and their derived subclones. Alate production in pea aphids was significantly increased in large populations of two PASS- and PAR-positive subclones relative to their parent clones. Attempts to transmit PASS or PAR horizontally, i.e., from aphid to aphid via feeding on host plants (bur clover), were unsuccessful.     

Phylogenetic relationships ofThiomicrospira species and their identification in deep-sea hydrothermal vent samples by denaturing gradient gel electrophoresis of 16S rDNA fragments

Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rDNA fragments was used to explore the genetic diversity of hydrothermal vent microbial communities, specifically to determine the importance of sulfur-oxidizing bacteria therein. DGGE analysis of two different hydrothermal vent samples revealed one PCR band for one sample and three PCR bands for the other sample, which probably correspond to the dominant bacterial populations in these communities. Three of the four 16S rDNA fragments were sequenced. By comparison with 16S rRNA sequences of the Ribosomal Database Project, two of the DGGE-separated fragments were assigned to the genusThiomicrospira. To identify these ‘phylotypes’ in more detail, a phylogenetic framework was created by determining the nearly complete 16S rRNA gene sequence (approx. 1500 nucleotides) from three describedThiomicrospira species, viz.,Tms. crunogena, Tms. pelophila, Tms. denitrificans, and from a new isolate,Thiomicrospira sp. strain MA2-6. AllThiomicrospira species exceptTms. denitrificans formed a monophyletic group within the gamma subdivision of the Proteobacteria.Tms. denitrificans was assigned as a member of the epsilon subdivision and was distantly affiliated withThiovulum, another sulfur-oxidizing bacterium. Sequences of two dominant 16S rDNA fragments obtained by DGGE analysis fell into the gamma subdivisionThiomicrospira. The sequence of one fragment was in all comparable positions identical to the 16S rRNA sequence ofTms. crunogena. Identifying a dominant molecular isolate asTms. crunogena indicates that this species is a dominant community member of hydrothermal vent sites. Another ‘phylotype’ represented a newThiomicrospira species, phylogenetically in an intermediate position betweenTms. crunogena andTms. pelophila. The third ‘phylotype’ was identified as aDesulfovibrio, indicating that sulfate-reducing bacteria, as sources of sulfide, may complement sulfur- and sulfide-oxidizing bacteria ecologically in these sulfide-producing hydrothermal vents.     

Effects of pea aphid secondary endosymbionts on aphid resistance and development of the aphid parasitoid Aphidius ervi: a correlative study

In order to reduce parasite‐induced mortality, hosts may be involved in mutualistic interactions in which the partner contributes to resistance against the parasite. The pea aphid, Acyrthosiphon pisum Harris (Hemiptera: Aphididae), harbours secondary bacterial endosymbionts, some of which have been reported to confer resistance against aphid parasitoids. Although this resistance often results in death of the developing parasitoid larvae, some parasitoid individuals succeed in developing into adults. Whether these individuals suffer from fitness reduction compared to parasitoids developing in pea aphid clones without symbionts has not been tested so far. Using 30 pea aphid clones that differed in their endosymbiont complement, we studied the effects of these endosymbionts on aphid resistance against the parasitoid Aphidius ervi Haliday (Hymenoptera: Braconidae: Aphidiinae), host–parasitoid physiological interactions, and fitness of emerging adult parasitoids. The number of symbiont species in an aphid clone was positively correlated with a number of resistance measurements but there were also clear symbiont‐specific effects on the host–parasitoid interaction. As in previous studies, pea aphid clones infected with Hamiltonella defensa Moran et al. showed resistance against the parasitoid. In addition, pea aphid clones infected with Regiella insecticola Moran et al. and co‐infections of H. defensa–Spiroplasma, R. insecticola–Spiroplasma, and R. insecticola–H. defensa showed reduced levels of parasitism and mummification. Parasitoids emerging from symbiont‐infected aphid clones often had a longer developmental time and reduced mass. The number of teratocytes was generally lower when parasitoids oviposited in aphid clones with a symbiont complement. Interestingly, unparasitized aphids infected with Serratia symbiotica Moran et al. and R. insecticola had a higher fecundity than unparasitized aphids of uninfected pea aphid clones. We conclude that in addition to conferring resistance, pea aphid symbionts also negatively affect parasitoids that successfully hatch from aphid mummies. Because of the link between aphid resistance and the number of teratocytes, the mechanism underlying resistance by symbiont infection may involve interference with teratocyte development.     

Phylotype diversity in a benthic cyanobacterial mat community on King George Island, maritime Antarctica

Cyanobacterial 16S ribosomal RNA gene diversity was examined in a benthic mat on Fildes Peninsula of King George Island (62o09′54.4′′S, 58o57′20.9′′W), maritime Antarctica. Environmental DNA was isolated from the mat, a clone library of PCR-amplified 16S rRNA gene fragments was prepared, and amplified ribosomal DNA restriction analysis (ARDRA) was done to assign clones to seven groups. Low cyanobacterial diversity in the mat was suggested in that 83% of the clones were represented by one ARDRA group. DNA sequences from this group had high similarity with 16S rRNA genes of Tychonema bourrellyi and T. bornetii isolates, whose geographic origins were southern Norway and Northern Ireland. Cyanobacterial morphotypes corresponding to Tychonema have not been reported in Antarctica, however, this morphotype was previously found at Ward Hunt Lake (83oN), and in western Europe (52oN). DNA sequences of three of the ARDRA groups had highest similarity with 16S rDNA sequences of the Tychonema group accounting for 9.4% of the clones. Sequences of the remaining three groups (7.6%) had highest similarity with 16S rRNA genes of uncultured cyanobacteria clones from benthic mats of Lake Fryxell and fresh meltwater on the McMurdo Ice Shelf.     

Endophytic Bacterial Diversity in Rice (Oryza sativa L.) Roots Estimated by 16S rDNA Sequence Analysis

The endophytic bacterial diversity in the roots of rice (Oryza sativa L.) growing in the agricultural experimental station in Hebei Province, China was analyzed by 16S rDNA cloning, amplified ribosomal DNA restriction analysis (ARDRA), and sequence homology comparison. To effectively exclude the interference of chloroplast DNA and mitochondrial DNA of rice, a pair of bacterial PCR primers (799f–1492r) was selected to specifically amplify bacterial 16S rDNA sequences directly from rice root tissues. Among 192 positive clones in the 16S rDNA library of endophytes, 52 OTUs (Operational Taxonomic Units) were identified based on the similarity of the ARDRA banding profiles. Sequence analysis revealed diverse phyla of bacteria in the 16S rDNA library, which consisted of alpha, beta, gamma, delta, and epsilon subclasses of the Proteobacteria, Cytophaga/Flexibacter/Bacteroides (CFB) phylum, low G+C gram-positive bacteria, Deinococcus-Thermus, Acidobacteria, and archaea. The dominant group was Betaproteobacteria (27.08% of the total clones), and the most dominant genus was Stenotrophomonas. More than 14.58% of the total clones showed high similarity to uncultured bacteria, suggesting that nonculturable bacteria were detected in rice endophytic bacterial community. To our knowledge, this is the first report that archaea has been identified as endophytes associated with rice by the culture-independent approach. The results suggest that the diversity of endophytic bacteria is abundant in rice roots.     

The use of visual cues in host evaluation by aphidiid wasps I. Comparison between threeAphidius parasitoids of the pea aphid

Host evaluation behaviour was examined in three species of aphid parasitoids,Aphidius ervi haliday,A. pisivorus Smith, andA. smithi Sharma & Subba Rao (Hymenoptera: Aphidiidae). Parasitoids were provided under laboratory conditions with three kinds of hosts representing two aphid species: (green) pea aphid,Acyrthosiphon pisum (Harris), and green and pink colour morphs of the alfalfa aphid,Macrosiphum creelii Davis. Females of all threeAphidius species distinguished between aphids on the basis of colour, movement, and host species. Patterns of host acceptance by parasitoids were species-specific. InA. ervi, host preference was the same in light and dark conditions: pea aphid>green alfalfa aphid≫pink alfalfa aphid. In contrast,A. pisivorus attacked and accepted pea aphid and green alfalfa aphid equally in the light and preferred both of these over pink alfalfa aphid; however, it made no distinction between pea aphid and pink alfalfa aphid in the dark. Females ofA. smithi attacked all three kinds of hosts (pea aphid>green alfalfa aphid≫pink alfalfa aphid) but apparently laid eggs only in pea aphid. The frequencies of attack and oviposition by all wasps were higher on ‘normal’ pea aphids than on those anaesthetized with CO₂. Host recognition is confirmed by chemical cues in the aphid cuticle that are detected during antennation, and host acceptance is dependent on an assessment of host quality during ovipositor probing.     

Diversity and geographic variation of endosymbiotic bacteria in natural populations of the pea aphid (Acyrthosiphon pisum) in China

Bacterial symbionts in aphids are known to benefit the insect host and associated with aphid’s ecological adaptation. The pea aphid (Acyrthosiphon pisum), an important legume pest worldwide, carries at least eight endosymbionts, providing a model system to study insect–bacteria interactions. However, species diversity and geographic variations of endosymbionts are unknown in Chinese populations; therefore, we characterized symbiont communities and diversity of 27 pea aphid samples from 13 geographic populations of China. Via amplicon high-throughput sequencing and diagnostic PCR, we found that bacterial communities of Chinese populations were dominated by Proteobacteria and Firmicutes. Among eight known endosymbionts, five (Buchnera, Serratia, Hamiltonella, Regiella, and Rickettsia) were detected by both methods, with a specific geographical distribution. The obligate symbiont, Buchnera, was present in all aphid samples, while the four facultative symbionts showed a significant geographic variation. Each population was randomly infected with distinct endosymbionts, ranging from three to five species. Serratia and Rickettsia showed relatively higher abundance in central regions of China, Regiella was predominant in eastern and western China, whereas Hamiltonella showed an extremely low abundance and was absent in four populations. Samples grouped by altitudes showed a significant diversity difference, whereas there was no significant difference between red and green body colors. Bacterial community structures of the Chinese pea aphid populations were mainly influenced by environmental factors, other than body colors. These data can guide the development of potential biocontrol techniques against this aphid.