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PubMed2003

Improvement of Barley yellow dwarf virus-PAV detection in single aphids using a fluorescent real time RT-PCR.

One of the major factors determining the incidence of Barley yellow dwarf virus (BYDV) on autumn-sown cereals is the viruliferous state of immigrant winged aphids. This variable is assessed routinely using the enzyme-linked immunosorbant assay (ELISA). However, the threshold for virus detection by ELISA can lead to false negative results for aphids carrying less than 10(6) particles. Although molecular detection techniques enabling the detection of lower virus quantities in samples are available, the relatively laborious sample preparation and data analysis have restricted their use in routine applications. A gel-free real-time one-step reverse transcription polymerase chain reaction (RT-PCR) protocol is described for specific detection and quantitation of BYDV-PAV, the most widespread BYDV species in Western Europe. This new assay, based on TaqMan technology, detects and quantifies from 10(2) to 10(8) BYDV-PAV RNA copies. This test is 10 and 10(3) times more sensitive than the standard RT-PCR and ELISA assays published previously for BYDV-PAV detection and significantly improves virus detection in single aphids. Extraction of nucleic acids from aphids using either phenol/chloroform or chelatin resin-based protocols allow the use of pooled samples or of a small part (up to 1/1600th) of a single aphid extract for efficient BYDV-PAV detection.

Animals↗
PubMed1997

Comparison of ELISA and RT-PCR for the detection of beet yellows closterovirus in plants and aphids.

A reverse-transcription polymerase chain reaction (RT-PCR) was developed to detect beet yellows closterovirus (BYV) in plants and single aphids using primers spanning the conserved regions of the published sequences of the coat protein gene and open reading frames 7 and 8. Three total RNA extraction procedures were examined and all were found to produce RNA of sufficient quality for RT-PCR, although the RNA extraction kit supplied by Flowgen was found to be the most versatile for the extraction of BYV from individual aphids. When 60 aphids, which had fed on virus infected sugar beet were tested by RT-PCR, 55% of individuals were found to contain BYV. Two groups of 36 individual aphids were tested by TAS-ELISA using a specific BYV monoclonal antibody; 53% gave positive absorbance values when a substrate amplification system was used, but none was found to contain virus when the system was replaced with the substrate p-nitrophenyl phosphate. An immunocapture RT-PCR method was shown to detect BYV regardless of whether the RNA had been extracted from the trapped particles or the reverse transcription and PCR mixtures were added to the wells containing intact particles. The RT-PCR method is now used to determine the numbers of BYV carrying aphids migrating into sugar-beet crops.

Animals↗
PubMed2000

Some epidemiological approaches to the control of aphid-borne virus diseases in seed potato crops in northern Europe.

An account is given of progress during the last 30 years in knowledge of the epidemiology of diseases caused by aphid-borne viruses in seed potato production areas of northern Europe. During this period, Potato leafroll virus (PLRV) and strains of Potato virus Y (PVY(O), PVY(N)) were consistently the most prevalent and harmful. The main factors influencing spread involve the amount of initial virus inoculum in seed crops, agricultural practices in relation to seed potato production, the status of aphids as colonising or transient vectors in the crop, variation in their life cycles and behaviour, effects of environmental factors on aphid population dynamics, and the interaction between aphid phenology and mature plant resistance. Lessons have been drawn from comparisons of the causes of outbreaks of PLRV and PVY, and various comprehensive models have been designed to predict virus spread and optimise control. The accuracy and reliability of virus detection, and chemical control of aphids have been considerably improved, but use of the latter has been severely affected by the development in Myzus persicae of resistance to most types of insecticide. It is suggested that more attention should be given to breeding virus-resistant cultivars that would lead to decreased pesticide use and thus promote more integrated environment-friendly strategies for control.

Animals↗
PubMed1996

Comparison of ecdysteroid concentration in different morphs of aphids.

The amounts of ecdysteroids were determined in different morphs of holocyclic monoecious aphids (Acyrthosiphon pisum Harris, Dysaphis devecta Walk., Lachnus roboris L., Schizolachnus pineti F.) and holocyclic heteroecious aphids (Aphis sambuci L., Rhopalosiphum padi L.) by means enzyme immunoassay. Among the parthenogenetic morphs (fundatrices, virginoparae, and oviparae), the fundatrices have consistently higher amounts of ecdysteroids than those of other morphs of the same species. Alate and apterous virginoparae showed slight differences in their ecdysteroid titer both in heteroecious and monoecious aphids. The migrant morphs (i.e., alate fundatrices and gynoparae) have the lowest amounts of ecdysteroids within a species. With the exception of D. devecta, the oviparae of both heteroecious and monoecious aphids have the second greatest amounts of ecdysteroids among the morphs living on same host plant. Polyphenism, dispersal behavior, and fecundity in connection with weight, ovariole number, and ecdysteroid concentration of different morphs of aphids are discussed.

Animals↗
PubMed2003

Identification of a gene overexpressed in aphids reared under short photoperiod.

Most aphids develop a cyclic parthenogenesis life-cycle. After several generations of viviparously produced parthenogenetic females, follows a single annual generation of sexual individuals, usually in autumn, that mate and lay the sexual eggs. Shortening of photoperiod at the end of the summer (together with temperature) is a key factor inducing the sexual response. Currently no genes involved in the cascade of events that lead to the appearance of sexual forms have been reported. After a Differential Display RT-PCR survey performed on Acyrthosiphon pisum aphids, we identified a gene that is overexpressed in aphids reared under short photoperiod conditions that induce sexuality in this species. This cDNA (called ApSDI-1) shows similarities with a protein involved in amino acid transport in GABAergic neurons. Since several studies implicate GABAergic transmission in the generation and modulation of circadian rhythmicity, we propose that ApSDI-1 could be involved in the transduction of the photoperiodic message and therefore be a candidate to participate at some point in processes that trigger the sexual response in aphids. This is the first gene identified in aphids whose expression is governed by the photoperiod.

Amino Acid Sequence↗
PubMed2003

Toxicity to the pea aphid Acyrthosiphon pisum of anti-chymotrypsin isoforms and fragments of Bowman-Birk protease inhibitors from pea seeds.

Aphids feed on a protein-poor diet and are insensitive to several serine protease inhibitors. However, among the Bowman-Birk family of plant trypsin inhibitors (BBI), some members display significant toxicity to the pea aphid Acyrthosiphon pisum. A BBI isoform purified from pea seeds (PsTI-2) displays an IC50 of 41 microM and a LC50 of 48 microM at 7 days. Our data show that the chymotrypsin-directed active site from these bifunctional inhibitors is responsible for this activity, and that artificial cyclic peptides bearing the Bowman-Birk anti-chymotrypsin head induce much greater toxicity and growth inhibition than their anti-trypsin counterparts. The toxic syndrome included a rapid behavioural response of aphids on diets containing the toxic peptides, with induced restlessness after only 1 h of exposure to the chymotrypsin inhibitor. Nevertheless, chymotrypsin activity was not detected in aphid guts, using two chromogenic chymotrypsin substrates, and the physiological target of the chymotrypsin inhibitor remains unknown. These data show for the first time that plant chymotrypsin inhibitors, still widely unexplored, may act as paradoxical toxicants to aphids and serve as defensive metabolites for phloem-feeding insects.

Amino Acid Sequence↗
PubMed2006

The genetic outcomes of sex and recombination in long-term functionally parthenogenetic lineages of Australian Sitobion aphids.

The typical life cycle of an aphid is cyclical parthenogenesis which involves the alternation of sexual and asexual reproduction. However, aphid life cycles, even within a species, can encompass everything on a continuum from obligate sexuality, through facultative sexuality to obligate asexuality. Loss of the sexual cycle in aphids is frequently associated with the introduction of a new pest and can occur for a number of environmental and genetic reasons. Here we investigate loss of sexual function in Sitobion aphids in Australia. Specifically, we aimed to determine whether an absence of sexual reproduction in Australian Sitobion results from genetic loss of sexual function or environmental constraints in the introduced range. We addressed our aims by performing a series of breeding experiments. We found that some lineages have genetically lost sexual function while others retain sexual function and appear environmentally constrained to asexuality. Further, in our crosses, using autosomal and X-linked microsatellite markers, we identified processes deviating from normal Mendelian segregation. We observed strong deviations in X chromosome transmission through the sexual cycle. Additionally, when progeny genotypes were examined across multiple loci simultaneously we found that some multilocus genotypes are significantly over-represented in the sample and that levels of heterozygosity were much higher than expected at almost all loci. This study demonstrates that strong biases in the transmission of X chromosomes through the sexual cycle are likely to be widespread in aphids. The mechanisms underlying these patterns are not clear. We discuss several possible alternatives, including mutation accumulation during periods of functional asexuality and genetic imprinting.

Animals↗
PubMed2003

Relationships between salicylic acid content, phenylalanine ammonia-lyase (PAL) activity, and resistance of barley to aphid infestation.

It has been suggested that salicylic acid (SA) is a signal in acquired resistance to pathogens in several plants. Also, it has been suggested that infestation of plants causes an increase in the activity of phenylalanine ammonia-lyase (PAL), a key phenolic biosynthesis enzyme. The purpose of this work was to investigate whether the induction of SA and PAL activity is related to the susceptibility of barley to aphid infestation. The induction of free and conjugated SA in two barley cultivars that differ in susceptibility to aphids was analyzed. Analyses of several physiological parameters showed that cv. UNA-80 was more susceptible to the aphid Schizaphis graminum than cv. LM-109. Salicylic acid was not detected in noninfested plants. Levels of free and conjugated SA in cv. LM-109 and of conjugated SA in cv. UNA-80 increased with aphid infestation, whereas the levels of free SA in cv. UNA-80 remained high under all infestation degrees. Maximum values reached in both cultivars were not significantly different. With respect to PAL activity, cv. LM-109 showed a significantly higher specific activity than cv. UNA-80, the more susceptible cultivar. The relationship between the susceptibility of a plant to aphid and SA induction and PAL activity is discussed.

Animals↗
PubMed2000

Identifying key cereal aphid predators by molecular gut analysis.

We describe polymerase chain reaction (PCR) primers for gut analysis of aphid predators. The primers amplify aphid mitochondrial COII fragments ranging in size from 77 to 386 bp. Using these primers, we were able to distinguish six species of US Great Plains cereal aphids, including two congeners, Rhopalosiphum maidis (Fitch) and R. padi (L.), and to detect them in extracts of coccinellid and chrysopid predators. We devised a protocol for deriving half-lives of detectability for the DNA of a single aphid consumed by predators maintained under simulated field dietary and temperature conditions. Using this protocol and primers that amplify a 198-bp fragment, we determined statistically different half-lives of detectability for a single R. maidis of 3.95 h in Chrysoperla plorabunda (Fitch) and 8. 78 h in Hippodamia convergens Guerin. The detectability half-life for a 339-bp R. maidis fragment was statistically longer in C. plorabunda but not in H. convergens. The sensitivity of the assay for the 198-bp fragment is 10-7 aphid equivalents. For species-specific predator gut analysis, PCR is superior to monoclonal antibody technology, giving comparable detectability half-lives with lower expense, much shorter development times, and greater certainty of a successful outcome.

Animals↗
PubMed2003

The evolution of inquilinism, host-plant use and mitochondrial substitution rates in Tamalia gall aphids.

We used mitochondrial DNA data to infer phylogenies for 28 samples of gall-inducing Tamalia aphids from 12 host-plant species, and for 17 samples of Tamalia inquilinus, aphid 'inquilines' that obligately inhabit galls of the gall inducers and do not form their own galls. Our phylogenetic analyses indicate that the inquilines are monophyletic and closely related to their host aphids. Tamalia coweni aphids from different host plants were, with one exception, very closely related to one another. By contrast, the T. inquilinus aphids were strongly genetically differentiated among most of their host plants. Comparison of branch lengths between the T. coweni clade and the T. inquilinus clade indicates that the T. inquilinus lineage evolves 2.5-3 times faster for the cytochrome oxidase I gene. These results demonstrate that: (1) Tamalia inquilines originated from their gall-inducing hosts, (2) communal (multi-female) gall induction apparently facilitated the origin of inquilinism, (3) diversification of the inquilines has involved rapid speciation along host-plant lines, or the rapid evolution of host-plant races, and (4) the inquilines have undergone accelerated molecular evolution relative to their hosts, probably due to reduced effective population sizes. Our findings provide insight into the behavioural causes and evolutionary consequences of transitions from resource generation to resource exploitation.

Animals↗
PubMed2002

Intracellular distribution of viral gene products regulates a complex mechanism of cauliflower mosaic virus acquisition by its aphid vector.

Interactions between Cauliflower mosaic virus (CaMV) and its aphid vector are regulated by the viral protein P2, which binds to the aphid stylets, and protein P3, which bridges P2 and virions. By using baculovirus expression of P2 and P3, electron microscopy, surface plasmon resonance, affinity chromatography, and transmission assays, we demonstrate that P3 must be previously bound to virions in order that attachment to P2 will allow aphid transmission of CaMV. We also show that a P2:P3 complex exists in the absence of virions but is nonfunctional in transmission. Hence, unlike P2, P3 and virions cannot be sequentially acquired by the vector. Immunogold labeling revealed the predominance of spatially separated P2:P3 and P3:virion complexes in infected plant cells. This specific distribution indicates that the transmissible complex, P2:P3:virion, does not form primarily in infected plants but in aphids. A model, describing the regulating role of P3 in the formation of the transmissible CaMV complex in planta and during acquisition by aphids, is presented, and its consequences are discussed.

Animals↗
PubMed2002

Influence of temperature on pea aphid Acyrthosiphon pisum (Hemiptera: Aphididae) resistance to natural enemy attack.

The ability to resist or avoid natural enemy attack is a critically important insect life history trait, yet little is understood of how these traits may be affected by temperature. This study investigated how different genotypes of the pea aphid Acyrthosiphon pisum Harris, a pest of leguminous crops, varied in resistance to three different natural enemies (a fungal pathogen, two species of parasitoid wasp and a coccinellid beetle), and whether expression of resistance was influenced by temperature. Substantial clonal variation in resistance to the three natural enemies was found. Temperature influenced the number of aphids succumbing to the fungal pathogen Erynia neoaphidis Remaudière & Hennebert, with resistance increasing at higher temperatures (18 vs. 28 degrees C). A temperature difference of 5 degrees C (18 vs. 23 degrees C) did not affect the ability of A. pisum to resist attack by the parasitoids Aphidius ervi Haliday and A. eadyi Starý, González & Hall. Escape behaviour from foraging coccinellid beetles (Hippodamia convergens Guerin-Meneville) was not directly influenced by aphid clone or temperature (16 vs. 21 degrees C). However, there were significant interactions between clone and temperature (while most clones did not respond to temperature, one was less likely to escape at 16 degrees C), and between aphid clone and ladybird presence (some clones showed greater changes in escape behaviour in response to the presence of foraging coccinellids than others). Therefore, while larger temperature differences may alter interactions between Acyrthosiphon pisum and an entomopathogen, there is little evidence to suggest that smaller changes in temperature will alter pea aphid-natural enemy interactions.

Animals↗
PubMed2004

Host plant age and population development of a cereal aphid, Metopolophium dirhodum (Hemiptera: Aphididae).

The sudden decline following the peak in population abundance of aphids on crops of small grain cereals is attributed to the joint effect of natural enemies and plant senescence. To distinguish between these causes, a four year experiment was established in which the numbers of Metopolophium dirhodum (Walker) infesting spring wheat plots sown from April to June at c. 14 day intervals were determined. Aphid abundance in replicates sown at successive dates peaked within a period of 5-9 days (106-171 day degrees above a base temperature of 0 degrees C) although their sowing dates varied by 62-97 days (727-1106 day degrees). At the time of the aphid population peaks, plants in the different sowings differed in age (11-99 days), developmental stage (stage 15-65 on the Zadoks scale), leaf nitrogen content and shoot mass. Maximum abundance of M. dirhodum decreased with sowing date because the time available for its population increase was shorter on late than early sowings. The abundance of M. dirhodum on spring wheat was similar to its abundance on winter wheat. After reaching peak abundance, aphids declined in numbers within 3-7 days. The effect of host plant ageing on the M. dirhodumdecline thus appeared small. Natural enemies (largely mycoses), and timing of alata production may have contributed to the aphid decline.

Animals↗
PubMed2006

Oviposition responses to patch quality in the larch ladybird Aphidecta obliterata (Coleoptera: Coccinellidae): effects of aphid density, and con- and heterospecific tracks.

The effects and persistence of oviposition-deterring semiochemical cues from conspecific and heterospecific larval tracks on the oviposition rate of Aphidecta obliterata (Linnaeus) females were investigated. In addition, the effects of varying aphid prey density were considered and also whether any resulting response originated from differential nutritional status of females and/or due to aphid odour stimuli. The existence of oviposition responses to conspecific egg chemicals was also considered. Gravid A. obliterata females were deterred from oviposition by conspecific larval tracks and the effect was density dependent. Females actively avoided searching in these contaminated areas. Tracks induced a significant effect on oviposition for up to three days. Heterospecific tracks of the coccinellid Adalia bipunctata (Fabricius) or the chrysopid Chrysoperla carnea (Stephens) did not induce any oviposition response in A. obliterata females. Increasing aphid density induced increased oviposition rate in A. obliterata females. Nutritional status of females was an important factor in the relationship between aphid density and oviposition rate, but aphid associated cues (odours) were not. There was an inhibitory effect of extracts of conspecific egg-surface chemicals on oviposition by A. obliterata females. In the field, cannibalism, competition and limited food availability represent the major threats to egg and larval survival. Patch quality assessment mechanisms enable females to lay eggs at sites where offspring survival is maximized. Oviposition-deterring semiochemicals tend to promote more even distribution of predators over prey patches.

Analysis of Variance↗
PubMed1994

Cloning and characterization of the ribosomal protein genes in the spc operon of a prokaryotic endosymbiont of the pea aphid, Acyrthosiphon kondoi.

To correlate a prokaryotic endosymbiont in the pea aphid, Acyrthosiphon kondoi, with the endosymbionts in related aphid species as well as with free-living bacteria and subcellular organelles, and to study the mode of its gene expression within aphid cells, we have cloned and characterized the genes encoding ribosomal proteins S3, L16, L29, S17, L14, L24, L5, S14, S8, L6, L18, S5, L30, L15 and secretion protein Y (Sec Y) from the S10 and spc ribosomal protein gene operons of this endosymbiont. The organization of these genes is identical to that in Escherichia coli, and their nucleotide sequences are highly similar (87% identity) to the corresponding E. coli genes. They are much less similar to the corresponding chloroplast and mitochondrial genes. The guanine plus cytosine G+C content of the genes of the A. kondoi endosymbiont is much higher than those of the endosymbionts in related aphid species reported so far. It appears either that the A. kondoi endosymbiont is derived from an ancestral bacterium different from those in other aphids or that its G+C content increased in a relatively short time after the evolutionary divergence of its host.

Amino Acid Sequence↗
PubMed2001

Intraspecific variation in symbiont genomes: bottlenecks and the aphid-buchnera association.

Buchnera are maternally transmitted bacterial endosymbionts that synthesize amino acids that are limiting in the diet of their aphid hosts. Previous studies demonstrated accelerated sequence evolution in Buchnera compared to free-living bacteria, especially for nonsynonymous substitutions. Two mechanisms may explain this acceleration: relaxed purifying selection and increased fixation of slightly deleterious alleles under drift. Here, we test the divergent predictions of these hypotheses for intraspecific polymorphism using Buchnera associated with natural populations of the ragweed aphid, Uroleucon ambrosiae. Contrary to expectations under relaxed selection, U. ambrosiae from across the United States yielded strikingly low sequence diversity at three Buchnera loci (dnaN, trpBC, trpEG), revealing polymorphism three orders of magnitude lower than in enteric bacteria. An excess of nonsynonymous polymorphism and of rare alleles was also observed. Local sampling of additional dnaN sequences revealed similar patterns of polymorphism and no evidence of food plant-associated genetic structure. Aphid mitochondrial sequences further suggested that host bottlenecks and large-scale dispersal may contribute to genetic homogenization of aphids and symbionts. Together, our results support reduced N(e) as a primary cause of accelerated sequence evolution in Buchnera. However, our study cannot rule out the possibility that mechanisms other than bottlenecks also contribute to reduced N(e) at aphid and endosymbiont loci.

Alleles↗
PubMed2003

Light activation of Russian wheat aphid-elicited physiological responses in susceptible wheat.

The impact of light and its role in Russian wheat aphid, Diuraphis noxia (Mordvilko), damage symptom formation, and photosynthetic capacity in 'Arapahoe' wheat (Triticum aestivum L.) were examined. After 72 h under continuous dark or continuous light regimes, the number of aphids (nymphs), leaf rolling and chlorosis ratings, fresh leaf weight, and chlorophyll contents were recorded. Photosynthetic rates, chlorophyll a, kinetics and chlorophyll extractions also were determined. Aphid infestation caused significant reductions in plant height, fresh weight, gas exchange, and chlorophyll fluorescence only under continuous light. Under the 72 h continuous dark regime, aphid infestation did not cause either damage symptom formation or reduction in plant growth or metabolism (photosynthesis). Furthermore, significantly more D. noxia nymphs were produced under continuous light condition than continuous dark. Our results demonstrate that the development of D. noxia feeding damage symptoms (i.e., leaf rolling and chlorotic streaks) on susceptible wheat seedlings is a light-activated process, even though the elicitor of the plant damage symptoms is aphid feeding.

Animals↗
PubMed2004

Enumerative and binomial sequential sampling plans for soybean aphid (Homoptera: Aphididae) in soybean.

Since the discovery of the soybean aphid, Aphis glycines Matsumura, in midwestern U.S. soybean, Glycine max L., in 2000, the aphid has become a significant economic pest. Basic information about estimating population density within fields is unknown. Therefore, we developed two sampling plans to efficiently characterize A. glycines densities. Enumerative and binomial sequential plans were developed using 89 data sets collected from 10 commercial fields sampled during 2001-2003. Re-sampling software was used to validate the enumerative plan on whole plant counts, based on Taylor's power law parameters (a = 9.157 and b = 1.543). For research applications, the enumerative plan was modified to provide an actual precision level of 0.10 (SE/mean), which resulted in an average sample number of 310 individual plants. For integrated pest management (IPM) purposes, we developed an enumerative plan with an actual precision of 0.25, which resulted in an average sample number of 38 individual plants. For IPM applications, the binomial plan will likely be more practical. Binomial plans were developed using two tally thresholds at five action thresholds. Final analysis of the operating characteristic curve for each plan indicated that the tally threshold of > or = 40 aphids per plant, and an action threshold of 0.837 (84% of the plants infested) provided the most correct treat (4%) and no-treat (95%) decisions, with very low incorrect treat (0.5%) and no-treat (0.5%) decisions. A tally threshold of > or = 40 aphids per plant and action thresholds of 84% of plants infested is equivalent to a mean density of 250 aphids per plant, a recently recommended economic threshold. Using this threshold, the minimum required sample number for the binomial plan was 11 plants.

Agriculture↗