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The secondary endosymbiotic bacterium of the pea aphid Acyrthosiphon pisum (Insecta: homoptera).

The secondary intracellular symbiotic bacterium (S-symbiont) of the pea aphid Acyrthosiphon pisum was investigated to determine its prevalence among strains, its phylogenetic position, its localization in the host insect, its ultrastructure, and the cytology of the endosymbiotic system. A total of 14 aphid strains were examined, and the S-symbiont was detected in 4 Japanese strains by diagnostic PCR. Two types of eubacterial 16S ribosomal DNA sequences were identified in disymbiotic strains; one of these types was obtained from the primary symbiont Buchnera sp., and the other was obtained from the S-symbiont. In situ hybridization and electron microscopy revealed that the S-symbiont was localized not only in the sheath cells but also in a novel type of cells, the secondary mycetocytes (S-mycetocytes), which have not been found previously in A. pisum. The size and shape of the S-symbiont cells were different when we compared the symbionts in the sheath cells and the symbionts in the S-mycetocytes, indicating that the S-symbiont is pleomorphic under different endosymbiotic conditions. Light microscopy, electron microscopy, and diagnostic PCR revealed unequivocally that the hemocoel is also a normal location for the S-symbiont. Occasional disordered localization of S-symbionts was also observed in adult aphids, suggesting that there has been imperfect host-symbiont coadaptation over the short history of coevolution of these organisms.

Animals↗

Interaction between the open reading frame III product and the coat protein is required for transmission of cauliflower mosaic virus by aphids.

Transmission of cauliflower mosaic virus (CaMV) by aphids requires two viral nonstructural proteins, the open reading frame (ORF) II and ORF III products (P2 and P3). An interaction between a C-terminal domain of P2 and an N-terminal domain of P3 is essential for transmission. Purified particles of CaMV are efficiently transmitted only if aphids, previously fed a P2-containing solution, are allowed to acquire a preincubated mixture of P3 and virions in a second feed, thus suggesting a direct interaction between P3 and coat protein. Herein we demonstrate that P3 directly interacts with purified viral particles and unassembled coat protein without the need for any other factor and that P3 mediates the association of P2 with purified virus particles. The interaction domain of P3 is located in its C-terminal half, downstream of the P3-P2 interaction domain but overlapping a region which binds nucleic acids. Mutagenesis of P3 which interferes with the interaction between P3 and virions is correlated with the loss of transmission by aphids. Taken together, our results demonstrate that P3 plays a crucial role in the formation of the CaMV transmissible complex by serving as a bridge between P2 and virus particles.

Animals↗

Resolving the aphid resistance locus Sd-1 on a BAC contig within a sub-telomeric region of Malus linkage group 7.

Aphids cause serious physical and economic damage to most major crops throughout the world, and there is a pressing requirement to isolate genes conferring aphid resistance. The Sd-1 locus in Malus spp. (apple) confers resistance against the rosy leaf-curling aphid (Dysaphis devecta Wlk.), and was recently positioned within a 1.3-cM region on linkage group 7, flanked by molecular markers. These markers were used as a basis for development of a BAC contig spanning the locus, together with adapter-mediated amplification of flanking sequences to obtain BAC insert-end sequences, and fingerprinting of BAC clones. Approximately 800 kb of the Sd-1 genomic region was covered by 19 overlapping BACs, with an average insert size of 75-150 kb. The physical-genetic distance ratio was estimated at 460 kb/cM, although the distribution of recombination events was irregular with respect to estimated physical distance. Recombinant analysis and development of new markers allowed Sd-1 to be positioned within an interval of approximately 180 kb located on either of two overlapping BACs. From one of these, an insert end sequence showed a significant degree of similarity to nucleotide binding site-leucine rich repeat (NBS-LRR) resistance genes. Fluorescent in situ hybridization (FISH) of BAC clones within the contig enabled positioning and orientation of the locus within a euchromatic region, very close to the telomere of linkage group 7.

Animals↗

Galling aphids: specialization, biological complexity, and variation.

Gall-inducing aphids are host specific in the gall stage. Most species alternate between trees (the primary host), where the gall is induced, and shrubs and grasses (secondary hosts). Parthenogenesis during most of their life cycle is interrupted by a single stage of sexual reproduction on the primary host. Apart from these general characteristics, galling aphids present some of the most complex and diverse life histories in the insect world. In this article I review the specialized characteristics of galling aphids, as well as their complex and diverse life histories, as reported in the past 20 years.

Adaptation, Physiological↗

Organization of the mitochondrial genomes of whiteflies, aphids, and psyllids (Hemiptera, Sternorrhyncha).

BACKGROUND: With some exceptions, mitochondria within the class Insecta have the same gene content, and generally, a similar gene order allowing the proposal of an ancestral gene order. The principal exceptions are several orders within the Hemipteroid assemblage including the order Thysanoptera, a sister group of the order Hemiptera. Within the Hemiptera, there are available a number of completely sequenced mitochondrial genomes that have a gene order similar to that of the proposed ancestor. None, however, are available from the suborder Sternorryncha that includes whiteflies, psyllids and aphids. RESULTS: We have determined the complete nucleotide sequence of the mitochondrial genomes of six species of whiteflies, one psyllid and one aphid. Two species of whiteflies, one psyllid and one aphid have mitochondrial genomes with a gene order very similar to that of the proposed insect ancestor. The remaining four species of whiteflies had variations in the gene order. In all cases, there was the excision of a DNA fragment encoding for cytochrome oxidase subunit III(COIII)-tRNAgly-NADH dehydrogenase subunit 3(ND3)-tRNAala-tRNAarg-tRNAasn from the ancestral position between genes for ATP synthase subunit 6 and NADH dehydrogenase subunit 5. Based on the position in which all or part of this fragment was inserted, the mitochondria could be subdivided into four different gene arrangement types. PCR amplification spanning from COIII to genes outside the inserted region and sequence determination of the resulting fragments, indicated that different whitefly species could be placed into one of these arrangement types. A phylogenetic analysis of 19 whitefly species based on genes for mitochondrial cytochrome b, NADH dehydrogenase subunit 1, and 16S ribosomal DNA as well as cospeciating endosymbiont 16S and 23S ribosomal DNA indicated a clustering of species that corresponded to the gene arrangement types. CONCLUSIONS: In whiteflies, the region of the mitochondrial genome consisting of genes encoding for COIII-tRNAgly-ND3-tRNAala-tRNAarg-tRNAasn can be transposed from its ancestral position to four different locations on the mitochondrial genome. Related species within clusters established by phylogenetic analysis of host and endosymbiont genes have the same mitochondrial gene arrangement indicating a transposition in the ancestor of these clusters.

Animals↗

Large-scale gene discovery in the pea aphid Acyrthosiphon pisum (Hemiptera).

Aphids are the leading pests in agricultural crops. A large-scale sequencing of 40,904 ESTs from the pea aphid Acyrthosiphon pisum was carried out to define a catalog of 12,082 unique transcripts. A strong AT bias was found, indicating a compositional shift between Drosophila melanogaster and A. pisum. An in silico profiling analysis characterized 135 transcripts specific to pea-aphid tissues (relating to bacteriocytes and parthenogenetic embryos). This project is the first to address the genetics of the Hemiptera and of a hemimetabolous insect.

Animals↗

Observation on the initial inoculum source and dissemination of Entomophthorales-caused epizootics in populations of cereal aphids.

A total number of 1092 migratory alates were trapped from air in wheat grown area of Yuanyang County, Henan Province from early April through May 2002 in order to confirm the source and dissemination of entomophthoralean inocula to cause epizootics of cereal aphids. Those included 415 Sitobion avenae, 642 Rhopalosiphum padi, 22 Metopolophium dirhodum, and 13 Schizaphis graminum. The trapped alates were daily collected and individually reared for 7 days on wheat plants in laboratory. Of those 341 alates died of fungal infection, taking 31.2% in the trapped alates. These included 224 S. avenae, 106 R. padi, 8 M. dirhodum, and 3 S. graminum. Deaths of all infected alates occurred during the first 5 days and 78.9% of the deaths occurred within the first 3 days. Individual examination under microscope proved that all deaths were attributed to entomophthoralean fungi. Of those Pandora neoaphidis accounted for 84.6%, Conidiobolus obscurus for 9.9%, and Entomophthora planchoniana for 5.5%. Four alate deaths died of cross infection of P. neoaphidis and C. conidiobolus. Based on the high infection rate of the migratory alates trapped from air and the field occurrence of epizootics in populations of cereal aphids during the trapping period, Entomophthorales-caused epizootics were likely disseminated by infected alates through their flight and colonization. This makes it reasonable to interpret worldwide distribution of aphid epizootics, particularly caused by P. neoaphidis that has no resting spores discovered.

Animals↗

Developmental origin and evolution of bacteriocytes in the aphid-Buchnera symbiosis.

Symbiotic relationships between bacteria and insect hosts are common. Although the bacterial endosymbionts have been subjected to intense investigation, little is known of the host cells in which they reside, the bacteriocytes. We have studied the development and evolution of aphid bacteriocytes, the host cells that contain the endosymbiotic bacteria Buchnera aphidicola. We show that bacteriocytes of Acyrthosiphon pisum express several gene products (or their paralogues): Distal-less, Ultrabithorax/Abdominal-A, and Engrailed. Using these markers, we find that a subpopulation of the bacteriocytes is specified prior to the transmission of maternal bacteria to the embryo. In addition, we discovered that a second population of cells is recruited to the bacteriocyte fate later in development. We experimentally demonstrate that bacteriocyte induction and proliferation occur independently of B. aphidicola. Major features of bacteriocyte development, including the two-step recruitment of bacteriocytes, have been conserved in aphids for 80-150 million years. Furthermore, we have investigated two cases of evolutionary loss of bacterial symbionts: in one case, where novel extracellular, eukaryotic symbionts replaced the bacteria, the bacteriocyte is maintained; in another case, where symbionts are absent, the bacteriocytes are initiated but not maintained. The bacteriocyte represents an evolutionarily novel cell fate, which is developmentally determined independently of the bacteria. Three of five transcription factors we examined show novel expression patterns in bacteriocytes, suggesting that bacteriocytes may have evolved to express many additional transcription factors. The evolutionary transition to a symbiosis in which bacteria and an aphid cell form a functional unit, similar to the origin of plastids, has apparently involved extensive molecular adaptations on the part of the host cell.

Animals↗

Glycoproteins from Russian wheat aphid infested wheat induce defence responses.

Elicitors are molecules which can induce the activation of plant defence responses. Elicitor activity of intercellular wash fluid from Russian wheat aphid, Diuraphis noxia (Mordvilko) infested resistant (cv Tugela DN), and susceptible (cv Tugela), wheat (Triticum aestivum L.), was investigated. Known Russian wheat aphid resistance related responses such as peroxidase and beta-1,3-glucanase activities were used as parameters of elicitor activity. The intercellular wash fluid from infested resistant plants contains high elicitor activity while that from infested susceptible plants contains no or very little elicitor activity. After applying C-18 reverse phase and concanavalin A Sepharose chromatography, elicitor active glycoproteins were isolated from the intercellular wash fluid of Russian wheat aphid infested resistant wheat. The elicitor-active glycoproteins separated into three polypeptides during sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isolated glycoproteins elicited peroxidase activity to higher levels in resistant than in susceptible cultivars. It was evident that the glycoproteins were probably a general elicitor of plant origin. Information gained from these studies is valuable for the development of plant activators to enhance the defence responses of plants.

Animals↗

Host selection, development, survival, and reproduction of turnip aphid (Homoptera: Aphididae) on green and red cabbage varieties.

Host selection, development, survival, and reproduction of the turnip aphid, Lipaphis erysimi (Kaltenbach), on selected green and red cabbage varieties, Brassicae oleracea L. variety capitata, were studied in the laboratory. Although numbers of alate L. erysimi on different varieties varied generally, more alate adults and nymphs were observed on green than on red varieties on day 3; whereas, these differences were not so significant on day 15. There were no significant differences in alate adults on all six varieties on day 15. Numbers of nymphs and apterous adults of L. erysimi varied among the varieties. When mean numbers of those aphids on green varieties were compared with those on red varieties, the green varieties had more nymphs and apterous adults than those on the red varieties. Developmental durations of L. erysimi nymphs also varied among all varieties, and the nymphs fed on green varieties developed longer than those fed on red varieties. Adult longevities on these cabbage varieties were not significantly different; whereas, the adult fecundities were. Survival rates of L. erysimi varied greatly among all varieties, but there are no significant differences between the green and red varieties. Factors that might affect aphid host selection, growth, longevity, fecundity, and survivorship were discussed.

Animals↗

Bionomics of Scymnus (Pullus) louisianae J. Chapin (Coleoptera: Coccinellidae) as a predator of the soybean aphid, Aphis glycines Matsumura (Homoptera: Aphididae).

A field population of Scymnus louisianae Chapin (Coccinellidae) was found attacking soybean aphids, Apis glycines Matsumura (Aphidae), a pest recently introduced into Kentucky. This coccinellid had not previously been found in Kentucky. A greenhouse population of S. louisianae was established and its predation on A. glycines studied under laboratory conditions. Total time to develop from egg to adult was about 20 d. About 70% of immatures survived to adulthood and they consumed approximately 100 aphid nymphs per beetle larva during the beetle's four larval instars. Adults lived for an average of 47 d (mated males) and 63 d (mated females) and, during their total adult lifetime, mated males consumed an average of 665 nymphs and mated females consumed 1261 nymphs. All developmental times and predation rates were comparable to those reported for other aphidophagous Scymnus spp. which, in conjunction with reports that Scymnus spp. are effective predators of cotton aphids, Aphis gossypii Glover, suggests that S. louisianae is a potentially important predator of A. glycines in the southern United States.

Animals↗

Fixed precision sequential sampling plans for the greenbug and bird cherry-oat aphid (Homoptera: Aphididae) in winter wheat.

The numbers of greenbugs, Schizaphis graminum (Rondani), and bird cherry-oat aphids, Rhopalosiphum padi L., per wheat tiller (stem) were estimated in 189 production winter wheat (Triticum aestivum L.) fields located throughout Oklahoma. Taylor's power law regressions were calculated from these data and used to construct fixed precision sequential sampling schemes for each species. An evaluation data set was constructed from 240 samples taken during three growing seasons from winter wheat fields at four locations in Oklahoma. Wheat cultivar and growth stage were recorded for each field on the day of sampling. Taylor's power law parameters for evaluation fields differed significantly for both species among growing seasons, locations, and plant growth stages. Median precision achieved using the fixed precision sequential sampling schemes for each species departed <20% from expected precision over the range population intensity in the evaluation data. For the 10% of samples with greatest deviation between observed and expected precision, observed precision was 13.8-81.8% greater than that expected precision depending on aphid species and population intensity. For the greenbug, the distribution of the percentage deviation between observed and expected precision was positively skewed, so that the sampling scheme tended to over-predict precision. For the bird cherry-oat aphid, the distribution was more symmetric. Even though precision observed using the sampling schemes frequently varied from expected precision, because of the inevitable consequence of sampling error and environmental variation, the sampling schemes yielded median observed precision levels close to expected precision levels over a broad range of population intensity.

Agriculture↗

Enzymatic chlorophyll degradation in wheat near-isogenic lines elicited by cereal aphid (Homoptera: Aphididae) feeding.

Chlorophyll degradation enzyme (i.e., chlorophyllase, Mg-dechelatase, and chlorophyll oxidase) activities of aphid-infested and uninfested 'Tugela' and Tugela near-isogenic wheat lines (i.e., Tugela-Dn1, Tugela-Dn2, and Tugela-Dn5) were assayed. Chlorophyllase activity was higher in bird cherry-oat aphid, Rhopalosiphum padi (L.) (Homoptera: Aphididae),-infested wheat lines compared with Russian wheat aphid, Diuraphis noxia (Mordvilko) (Homoptera: Aphididae)]-infested and uninfested plants. Mg-dechelatase activity was higher in D. noxia-infested wheat lines than in R. padi-infested and uninfested plants. Also, Mg-dechelatase activity was lower in Tugela wheat infested with D. noxia than in Tugela near-isogenic lines with Dn genes. Based on the in vitro assays of chlorophyll degradation enzyme (i.e., chlorophyllase and Mg-dechelatase) activities, we proposed that the chlorotic symptoms observed on D. noxia-infested Tugela wheat were most likely to be elicited by unbalanced chlorophyll biosynthesis and degradation.

Animals↗

Yield response and categories of resistance to Russian wheat aphid in four Dn4 hard red winter wheat cultivars.

A field experiment was conducted to determine whether resistance to Russian wheat aphid, Diuraphis noxia (Mordvilko), conferred by the Dn4 gene is affected by genetic background. This was done by comparing the yield responses to Russian wheat aphid-resistant wheat containing Dn4, derived through the backcross method, to those of the corresponding recurrent parents. Infested resistant cultivars had fewer Russian wheat aphids per tiller than infested susceptible cultivars at the Lamar and Fort Collins, CO sites but not at the Akron, CO site. At the Lamar site, resistant cultivars yielded more than the susceptible cultivars. 'Prairie Red' and 'Yumar' were more resistant than 'Prowers', especially at the higher infestation level. Resistance in these cultivars was categorized in a laboratory experiment to confirm this differential expression of resistance. Resistance in Prairie Red, 'Halt', 'Prowers 99', and Yumar was categorized at three plant growth stages. Antibiosis was expressed as reductions in maximum number of nymphs produced per 24 h and intrinsic rate of increase. The maximum number of nymphs produced per 24 h was reduced in Halt and 'Lamar'. Averaged over cultivars, the intrinsic rate of increase was less at jointing than at the seedling or tillering growth stages. Tolerance was expressed in the resistant cultivars as reduced chlorosis and leaf rolling. Growth reductions in infested Prowers 99 plants was less than the other cultivars. This study confirms that some cultivars containing Dn4 may express antibiosis and tolerance, whereas others may not show the same categories. Thus, expression is affected by genetic background.

Animals↗

Attributes of bean yellow mosaic potyvirus transmission from clover to snap beans by four species of aphids (Homoptera: Aphididae).

After characterization of the natural spread of necrosis-inducing Bean yellow mosaic potyvirus (family Potyviridae, genus Potyvirus, BYMV(N)), nonpersistently transmitted from clover, Trifolium repens L., to an adjacent field of snap bean, Phaseolus vulgaris L., in western Oregon, we established a study site enabling us to investigate the virus reservoir, to observe en masse transmission of BYMV(N) to bean plants, and to identify aphid species associated with virus spread. Colonies of Myzus persicae (Sulzer), Acyrthosiphon pisum (Harris), and Aphis fabae Scopoli associated with virus spread were established in an insectary and shown to vector this virus. Although Nearctaphis bakeri (Cowen) comprised 68% of aphid alatae taken from bean leaves during virus spread, we were unable to show that this species could vector the virus by using the same methods that were successful for the other species. Instead, we found that when two distinct N. bakeri colonies unexpectedly emerged from the roots of T. repens BYMV(N) source plants (WZwc #6 and #11) that were present in the laboratory (insectary), these aphids transmitted BYMVN at rates comparable with those of M. persicae and A. pisum. Transmission of BYMVN also occurred with two other N. bakeri colonies maintained for 4 mo on Trifolium pratense L. (NZwc Sch 3B and Sch 7C) BYMVN source plants. Each of these four BYMVN transmission successes also demonstrated an unprecedented once-only transmission of BYMV(N) by N. bakeri colonies. Our experience with western Oregon N. bakeri colonies was compared with descriptions of this native North American species after its 1960-1980s arrival in France, Germany, and Italy.

Animals↗

Characterization of antibiosis and antixenosis to the soybean aphid (Hemiptera: Aphididae) in several soybean genotypes.

The soybean aphid, Aphis glycines Matsumura (Hemiptera: Aphididae), is an introduced pest of soybean, Glycine max (L.) Merr., in North America, and it can reduce yields by 50%. Since 2000, when A. glycines was first detected in the United States, studies of this insect and possible control methods have been initiated. Plant resistance to this aphid species is one important component of integrated control. Reproduction of A. glycines was compared on 240 soybean entries in a pesticide-free greenhouse. Eleven entries had fewer nymphs produced, compared with the susceptible checks, and these entries were used in follow-up experiments to assess antibiosis and antixenosis. Antibiosis was estimated in true no-choice tests, in which adults were confined individually in double-sided sticky cages stuck to the upper side of leaves. Antixenosis was assessed in choice tests, in which all entries were planted in a single pot. Adult aphids were placed in the center of the pot, and 24 h later the number of adults on each plant was counted. Of the 11 entries evaluated, nine showed a moderate antibiotic effect to A. glycines, and the other two entries (K1639 and Pioneer 95B97) showed not only a strong antibiotic effect but also exhibited antixenosis as a category of resistance to A. glycines. The resistant soybean entries found in this work are potential sources for A. glycines control.

Animals↗

Quantitative assessment of the infection rate of the entomophthoraceous fungus, Zoophthora anhuiensis against the green peach aphid Myzus persicae.

A two-step method was developed to quantitatively assess the infection rate of the entomophthoraceous fungus, Zoophthora anhuiensis (Li) Humber, on the green peach aphid, Myzus persicae (Sulzer). Firstly, a standard time-dose-mortality relationship, established by modeling data from bioassay 1 at varying conidial dosages (0.4 - 10.4 conidia/mm2) of Z. anhuiensis F97028, was used to yield an estimate of expected mortality probability at a given dosage. Secondly, bioassay 2 was conducted by simultaneously exposing six < or = 4-day-old nymphal colonies to a shower of Z. anhuiensis conidia at each of four dosages (resulting from exposures of 0.3 - 8.0 min). Subsequently, the colonies were separately immersed in a 0.1% chlorothalonil solution for 0.5 min to disinfect all surviving conidia on the host integument from 1 - 12 h after exposure under temperature treatments of 15 and 20 degrees C, respectively. The infection rate during a specific period from the end of the exposure to the immersion was then estimated as the ratio of the observed mortality over the expected mortality probability at a particular dosage. The results showed that the infection of M. persicae from Z. anhuiensis was highly rapid with little difference between aphid colonies maintained at 15 and 20 degrees C before being immersed in the fungicidal solution after exposure. The first 6-hour period after exposure was most crucial to successful infection of the fungus with the infection rate greatly depending on conidial dosages. It took < or = 1 h to infect > 50% of the aphids at a dosage of > 1.5 conida/mm2 and > 90% at > 50 conidia/mm2.

Animals↗

Rapidly evolving aphid gall effector proteins exhibit saposin-like folds.

Many insects manipulate plants by injecting effector proteins. In one extreme example of this molecular "hijacking", Hormaphis cornu aphids inject bicycle proteins into Hamamelis virginiana (Witch Hazel), contributing to the development of novel organs called galls. Bicycle proteins share no amino acid sequence similarity with proteins of known function. Here, we report the crystal structures of two divergent bicycle proteins. Both proteins contain saposin-like folds: one with multiple disulfide bonds exhibits a helix swap; the other has no disulfide bonds and possesses two tandem domains. To explore the structural evolution of bicycle proteins, we predicted bicycle protein structures with Alphafold2 (AF2). While AF2 did not recover the two experimental structures using existing databases, it succeeded after we provided multiple sequence alignments (MSAs) containing protein sequences encoded in new genome sequences from closely related aphid species. Using this customized approach at scale, we generated 2400 high-confidence predictions for bicycle proteins from seven aphid species. This dataset revealed that bicycle proteins without cysteines are outliers in fold space and appear to have evolved from ancestral proteins with disulfide-bonded saposin-like folds. While all bicycle proteins contain predicted saposin-like folds, they display a vast diversity of structural and physicochemical properties. While this diversity thwarts prediction of conserved functions encoded in structure, it suggests that bicycle proteins have evolved to target diverse plant processes and/or to evade plant immune surveillance.

AlphaFold predictions↗