Parasitoids and biocontrol of Russian wheat aphid, Diuraphis noxia (Kurdj.) expanding in central Europe

Hymenopterous parasitoids found on the Russian wheat aphid (RWA), Diuraphis noxia (Kurdj.) since the aphid was detected in the Czech Republic in 1995 have been studied. Parasitoids were obtained from aphid samples taken in small grains and from field exposures of banker-units containing laboratory mass-reared RWA. The parasitoid species are listed. Their presence on RWA was verified and their host species alternation was also tested in the laboratory. Evidence on some species was obtained only through unnatural host propagation. Aphidius colemani Vier. (a Chilean strain) was purposely introduced, released and became established; its host range, which included both target and non-target species, is listed. Prediction versus determination of RWA parasitoid complex in central Europe are discussed and the field samples and the exposure trials are compared. The RWA presence in relation to biodiversity in the small-grains agroecosystem is analysed. An overall parasitoid spectrum of RWA in the Old World (and the world background) is reviewed in relation to the RWA native range and the distribution (expansion) range, respectively. A corresponding analysis of the central European situation, derived from the expansion route of RWA to the Mediterranean, is presented.     

In vitro selection for Russian wheat aphid (Diuraphis noxia) resistance in wheat (Triticum aestivum)

The Russian wheat aphid (Diuraphis noxia) is a pest on wheat (Triticum aestivum) in many regions of the world. The aphid injects a phytotoxin when it feeds. Identification of somaclonal variants with phytotoxin resistance may shorten development time for resistance. Wheat calli from the susceptible cultivar Stephens were exposed to an extract from the aphid. Five plants were regenerated from 100 treated calli. Resistance to the aphid was observed in both the R₂ and R₃ generations. One of the six R₃ populations had improved resistance for leaf curling and leaf folding, while another had improved response for chlorosis damage. These results indicate that the use of aphid extract on wheat callus offers an alternate method for development of resistance to the Russian wheat aphid.     

Antioxidative enzymes and the Russian wheat aphid (Diuraphis noxia) resistance response in wheat (Triticum aestivum)

A crucial function of antioxidative enzymes is to remove excess reactive oxygen species (ROS), which can be toxic to plant cells. The effect of Russian wheat aphid (RWA), Diuraphis noxia (Mordvilko), infestation on the activities of antioxidative enzymes was investigated in the resistant (cv. Tugela DN) and the near-isogenic susceptible (cv. Tugela) wheat (Triticum aestivum L.). RWA infestation significantly induced the activity of superoxide dismutase, glutathione reductase and ascorbate peroxidase to higher levels in the resistant than in susceptible plants. These findings suggest the involvement of antioxidative enzymes in the RWA-wheat resistance response, which was accompanied by an early oxidative burst. The results are consistent with the role of ROS in the resistance response and the control of their levels to minimise toxic effects.     

Supercooling capacity of Eurasian and North American populations of parasitoids of the Russian wheat aphid, >Diuraphis noxia

Supercooling points were estimated for seven populations of >Aphelinus albipodus, five populations of >Aphelinus asychis, and four populations of >Diaeretiella rapae to assess whether their supercooling points were sufficiently low to provide the potential for overwintering survival in colder temperate climatic areas. Test individuals from all 16 of the parasitoid populations were collected originally from mummies of the Russian wheat aphid, >Diuraphis noxia. Mummies containing parasitoid pupae were maintained for 1 wk under three different temperature conditions (treatments): at room temperature (24.8 ± 0.2 °C), 1 wk at 0 °C, and 1 wk –5 °C, and the supercooling points across treatments, and within and among species were compared. Statistical differences in supercooling points were found among populations of >A. albipodus for each treatment, and for >A. asychis when maintained for 1 wk at room temperature. No differences in supercooling points were found among populations of >D. rapae mummies maintained under the three temperature treatments. The lowest supercooling points obtained for the three parasitoid species maintained at room temperature were the >A. albipodus population from Montana (–31.68 °C), the >A. asychis population from Greece (–32.04 °C), and the >D. rapaepopulation from the Caucasus (–33.12 °C). Preconditioning the parasitoid mummies to cold had no effect on the supercooling points for >A. albipodus, and in some cases unexpectedly increased the supercooling points for >A. asychisand >D. rapae. In comparing the overall mean supercooling points of the three parasitoid species, no differences were found within species (among temperature treatments), nor among species (within temperature treatments). It was concluded that observed differences in supercooling points of only a few degrees Centigrade among parasitoid populations and species would not be expected to cause differences in their overwintering success, especially given the expected variability in temperatures within and among overwintering sites.     

Proteomic analysis of secreted saliva from Russian Wheat Aphid (Diuraphis noxia Kurd.) biotypes that differ in virulence to wheat

Diuraphis noxia, Russian Wheat Aphid (RWA), biotypes are classified by their differential virulence to wheat varieties containing resistance genes. RWA salivary proteins, unlike those of most aphid species, cause foliar damage and physiological alterations in plants. A comparative proteomic analysis of secreted saliva from four differentially virulent RWA biotypes identified thirty-four individual proteins. The five major proteins were glucose dehydrogenase, lipophorin, chitinase, CiV16.8g1-like, and lava lamp. Fourteen proteins quantitatively varied among biotypes; trehalase, β-N-acetylglucosaminidase (chitinase), two separate glucose dehydrogenases, calreticulin, aminopeptidase, acetylglucosaminyltransferase, hydroxymethylglutaryl-CoA lyase, acyltransferase, ficolin-3, lava lamp, retinaldehyde-binding protein, and two proteins of unknown function. Fifty-four percent of spectral counts were associated with glucose dehydrogenase, which is thought to detoxify plant defensive compounds. One-dimensional electrophoresis detected nine protein bands from 9 to 60 kDa that quantitatively differed. Two-dimensional electrophoresis identified six major gel zones with quantitative and qualitative variance in proteins. Our findings reveal that the salivary proteome of RWA, a phytotoxic aphid, differs considerably from those reported for nonphytotoxic aphids. The potential roles of proteins used in the general plant feeding processes of aphids and those that are potential phytotoxins related to aphid virulence are discussed.     

A proteomic evaluation of Pyrenophora tritici‐repentis,causal agent of tan spot of wheat,reveals major differences between virulent and avirulent isolates

Pyrenophora tritici‐repentis causes tan spot, an important foliar disease of wheat. The fungus produces multiple host‐specific toxins, including Ptr ToxB, a chlorosis‐inducing protein encoded by the ToxB gene. A homolog of ToxB is also found in avirulent isolates of the fungus. In order to improve understanding of the role of this homolog and evaluate the general pathogenic ability of P. tritici‐repentis, we compared the proteomes of avirulent race 4 and virulent race 5 isolates of the pathogen. Western blotting analysis revealed the presence of Ptr ToxB in spore germination and culture fluids of race 5 but not race 4. A comprehensive proteome‐level comparison by 2‐DE indicated 133 differentially abundant proteins in the secretome (29 proteins) and mycelium (104 proteins) of races 4 and 5, of which 63 were identified by MS/MS. A number of the proteins found to be up‐regulated in race 5 have been implicated in microbial virulence in other pathosystems, and included the secreted enzymes α‐mannosidase and exo‐β‐1,3‐glucanase, heat‐shock and BiP proteins, and various metabolic enzymes. These proteome‐level differences suggest a reduced general pathogenic ability in race 4 of P. tritici‐repentis, irrespective of toxin production. Such differences may reflect an adaptation to a saprophytic habit.     

Comparison of categories of resistance in wheat and barley genotypes against biotype 2 of the Russian wheat aphid, Diuraphis noxia (Kurdjumov)

The Russian wheat aphid Diuraphis noxia (Kurdjumov) (Homoptera: Aphididae) is a global pest of wheat and barley. This arthropod is difficult to manage with pesticides or biological control agents due to the aphid’s ability to seek shelter in rolled leaves and also to develop virulent biotypes. During the past 20 years, the use of aphid-resistant cereal cultivars has proven to be an economically and ecologically beneficial method of protecting crops from D. noxia damage. Our research reports the results of experiments to determine the categories of D. noxia biotype 2 resistance present in Cereal Introduction Triticeae (CItr) 2401, and a barley genotype (IBRWAGP4-7), compared to control resistant and susceptible wheat and barley genotypes. CItr2401 and IBRWAGP4-7 exhibit no antixenosis, but both genotypes demonstrated antibiosis to D. noxia in the form of reduced aphid populations. Reduced leaf dry weight change, a measure of plant tolerance of D. noxia feeding, was significantly less in CItr2401 and IBRWAGP4-7 plants than in plants of susceptible control varieties. However, tolerance was negated when a tolerance index was calculated to correct for differences in aphid populations. Barley IBRWAGP4-7 is a new source of D. noxia biotype 2 resistance. D. noxia foliar leaf damage and population growth were significantly less on IBRWAGP4-7 plants than on plants of the susceptible barley variety Morex. IBRWAGP4-7 plants were equal in resistance to plants of the resistant barley STARS 9301 and wheat genotype CItr2401. Handling editor: Heikki Hokkanen     

Comparison of Diuraphis noxia resistance in wheat isolines and plant introduction lines

A bioassay to examine the foraging behaviour of the aphidophagous hoverfly, Episyrphus balteatus was used in which various stimuli purported to influence flower choice were tested using model flowers. In choice experiments, E. balteatus exhibited enhanced behavioural responses to the colour yellow, as previously seen in Eristalis by several workers. Previous suggestions that the generalist forager E. balteatus has no preference for colour are re-examined in the light of this new evidence. The other advertisement cue tested, size, was also demonstrated to play an important role in determining foraging behaviour, with the smaller artificial flowers seemingly preferred. Of the rewards tested, E. balteatus showed preference for the greatest nectar concentrations, whilst increasing pollen did not affect behaviour. These bioassays provided an opportunity to isolate the individual components of decision-making by E. balteatus during foraging. The separation of sexes and ages in these experiments permitted analysis of inter-sexual and inter-generation differences in behaviour, a factor apparently not investigated in previous work.     

Characteristics of the population build-up of the Russian wheat aphid Diuraphis noxia and the effect on wheat yield in the eastern Orange Free State

Diuraphis noxia infestations were monitored in early winter and winter wheat during 1983–1985. The aphid population increase was logarithmic in all three years and on both cultivars 100 % infestation levels were recorded. The period of exponential increase in both percentage infestation and in the number of D. noxia per stem took place when the plants had reached the stem elongation stage. The mean number recorded at peak population levels was 150–160 aphids per stem. During the period of plant senescence numbers dropped rapidly. Large reductions in yield were recorded on all infested plants. Single-plant data showed that yield reductions in the early winter cv. Betta were related to the duration of infestation. Relative to sprayed controls, reduction in kernel mass ranged from 25–80% and in 1000-kernel mass from 15–45%. However, the yield reductions of the faster-growing cv. Flamink, appeared to be dependent on the number of aphids per plant during the later growth stages and not the duration of infestation.     

Comparative whole-genome analysis of virulent and avirulent strains of Porphyromonas gingivalis

We used Porphyromonas gingivalis gene microarrays to compare the total gene contents of the virulent strain W83 and the avirulent type strain, ATCC 33277. Signal ratios and scatter plots indicated that the chromosomes were very similar, with approximately 93% of the predicted genes in common, while at least 7% of them showed very low or no signals in ATCC 33277. Verification of the array results by PCR indicated that several of the disparate genes were either absent from or variant in ATCC 33277. Divergent features included already reported insertion sequences and ragB, as well as additional hypothetical and functionally assigned genes. Several of the latter were organized in a putative operon in W83 and encoded enzymes involved in capsular polysaccharide synthesis. Another cluster was associated with two paralogous regions of the chromosome with a low G+C content, at 41%, compared to that of the whole genome, at 48%. These regions also contained conserved and species-specific hypothetical genes, transposons, insertion sequences, and integrases and were located adjacent to tRNA genes; thus, they had several characteristics of pathogenicity islands. While this global comparative analysis showed the close relationship between W83 and ATCC 33277, the clustering of genes that are present in W83 but divergent in or absent from ATCC 33277 is suggestive of chromosomal islands that may have been acquired by lateral gene transfer.     

A multi‐genome analysis approach enables tracking of the invasion of a single Russian wheat aphid (Diuraphis noxia) clone throughout the New World

This study investigated the population genetics, demographic history and pathway of invasion of the Russian wheat aphid (RWA) from its native range in Central Asia, the Middle East and Europe to South Africa and the Americas. We screened microsatellite markers, mitochondrial DNA and endosymbiont genes in 504 RWA clones from nineteen populations worldwide. Following pathway analyses of microsatellite and endosymbiont data, we postulate that Turkey and Syria were the most likely sources of invasion to Kenya and South Africa, respectively. Furthermore, we found that one clone transferred between South Africa and the Americas was most likely responsible for the New World invasion. Finally, endosymbiont DNA was found to be a high‐resolution population genetic marker, extremely useful for studies of invasion over a relatively short evolutionary history time frame. This study has provided valuable insights into the factors that may have facilitated the recent global invasion by this damaging pest.     

Russian Wheat Aphid,Diuraphis noxia in the Czech Republic – cause of the significant population decrease

Diuraphis noxia is a newly established alien pest in the Czech Republic. There were two population explosions during the period 1993–2012. A successful overwintering of the anholocyclic populations of D. noxia was the cause of a population explosion in the oncoming growth season. Viviparous parthenogenetic females of the anholocyclic populations and diapausing eggs of holocyclic populations could overwinter at the same time. Analysis of the course of winter temperatures showed that a complete winter mortality of anholocyclic populations depended mainly on the duration of the freezing period, when temperature dropped below ?5°C, and on the intensity of the frost. We proposed the cumulative effective freeze (in minus degree days) as a key parameter for the aphid anholocyclic population's mortality during overwintering. This parameter equalled ?10.1 degree days for relatively warm winter period 2006–2007, whereas in the relatively cold winter period of 2009–2010, this parameter reached ?87.1 DD, which caused complete mortality of anholocyclic populations. A cumulative effective freeze could be used for short‐time prognosis of D. noxia population explosion.     

Genome-wide transcriptome analysis of porcine epidemic diarrhea virus virulent or avirulent strain-infected porcine small intestinal epithelial cells

Porcine epidemic diarrhea virus (PEDV) is the main cause of diarrhea, vomiting, and mortality in pigs, which results in devastating economic loss to the pig industry around the globe. In recent years, the advent of RNA-sequencing technologies has led to delineate host responses at late stages of PEDV infection; however, the comparative analysis of host responses to early-stage infection of virulent and avirulent PEDV strains is currently unknown. Here, using the BGI DNBSEQ RNA-sequencing, we performed global gene expression profiles of pig intestinal epithelial cells infected with virulent (GDS01) or avirulent (HX) PEDV strains for 3, 6, and 12 h. It was observed that over half of all significantly dysregulated genes in both infection groups exhibited a down-regulated expression pattern. Functional enrichment analyses indicated that the differentially expressed genes (DEGs) in the GDS01 group were predominantly related to autophagy and apoptosis, whereas the genes showing the differential expression in the HX group were strongly enriched in immune responses/inflammation. Among the DEGs, the functional association of TLR3 and IFIT2 genes with the HX and GDS01 strains replication was experimentally validated by TLR3 inhibition and IFIT2 overexpression systems in cultured cells. TLR3 expression was found to inhibit HX strain, but not GDS01 strain, replication by enhancing the IFIT2 expression in infected cells. In conclusion, our study highlights similarities and differences in gene expression patterns and cellular processes/pathways altered at the early-stage infection of PEDV virulent and avirulent strains. These findings may provide a foundation for establishing novel therapies to control PEDV infection.     

Comparative analysis of glycoprotein and glycolipid composition of virulent and avirulent strain membranes ofMycoplasma hyopneumoniae

The glycoproteins and glycolipids from membranes of virulent strain Z and avirulent strain M ofMycoplasma hyopneumoniae have been compared. The proteins and the glycoproteins were identified by SDS-polyacrylamide gel electrophoresis and concanavalin A-biotin labeling, respectively. The membrane preparation contained approximately 34 protein bands with molecular weights between 20 KD and 100 KD. The concanavalin A-biotin system reacted with a glycoprotein of a molecular weight of approximately 28,000 from avirulent strain M and did not react with the correspondent band from virulent strain Z. The membrane glycolipids of both strains consisted of monogalactosyldiacylglycerol (MGDG) and digalactosyldiacylglycerol (DGDG), and the percentages of 160, 180, and 181 fatty acids comprised more than 80% of the total fatty acids of membrane glycolipids. The 180 fatty acid of MGDG in avirulent strain M was twofold higher than that of virulent strain Z.     

Comparative transcriptome analysis of Listeria monocytogenes strains of the two major lineages reveals differences in virulence, cell wall, and stress response

Listeria monocytogenes is a food-borne, opportunistic, bacterial pathogen causing a wide spectrum of diseases, including meningitis, septicemia, abortion, and gastroenteritis, in humans and animals. Among the 13 L. monocytogenes serovars described, human listeriosis is mostly associated with strains of serovars 4b, 1/2b, and 1/2a. Within the species L. monocytogenes, three phylogenetic lineages are described. Serovar 1/2a belongs to phylogenetic lineage I, while serovars 4b and 1/2b group in phylogenetic lineage II. To explore the role of gene expression in the adaptation of L. monocytogenes strains of these two major lineages to different environments, as well as in virulence, we performed whole-genome expression profiling of six L. monocytogenes isolates of serovars 4b, 1/2b, and 1/2a of distinct origins, using a newly constructed Listeria multigenome DNA array. Comparison of the global gene expression profiles revealed differences among strains. The expression profiles of two strains having distinct 50% lethal doses, as assessed in the mouse model, were further analyzed. Gene ontology term enrichment analysis of the differentially expressed genes identified differences in protein-, nucleic acid-, carbon metabolism-, and virulence-related gene expression. Comparison of the expression profiles of the core genomes of all strains revealed differences between the two lineages with respect to cell wall synthesis, the stress-related sigma B regulon and virulence-related genes. These findings suggest different patterns of interaction with host cells and the environment, key factors for host colonization and survival in the environment.     

Comparative assessment of acid, alkali and salt tolerance in Listeria monocytogenes virulent and avirulent strains

Listeria monocytogenes is an opportunistic bacterial pathogen of man and animals that has the capacity to survive under extreme environmental conditions. While our knowledge on L. monocytogenes and its ability to sustain within wide pH and temperature ranges and salt concentrations has been largely built on the virulent strains of this species, relatively little is known about avirulent strains in this regard. In this study, we extend our analysis on avirulent L. monocytogenes strains. By subjecting three virulent (EGD, 874 and ATCC 19196) and three avirulent (ATCC 19114, HCC23 and HCC25) strains to various pH and salt concentrations, it was found that L. monocytogenes recovered well after treatment with 100 mM Tris at pH 12.0, and to a lesser extent at pH 3.0. Interestingly, avirulent L. monocytogenes strains showed a somewhat higher tolerance to alkali than virulent strains. This unique feature of avirulent L. monocytogenes strains may potentially be exploited for the development of a rapid technique for differentiation between avirulent and virulent strains. Furthermore, all L. monocytogenes strains tested were resistant to saturated NaCl (about 7 M, or 40% w/v) for a long period of time (20 h and possibly longer). Together, these results highlight that acid, alkali, and/or salt treatments commonly used in food product processing may not be sufficient to eliminate L. monocytogenes, and therefore stringent quality control measures at the beginning and end of the food manufacturing process is essential to ensure that such food products are free of listerial contamination.     

Comparative DNA sequence analysis of mapped wheat ESTs reveals the complexity of genome relationships between rice and wheat

The use of DNA sequence-based comparative genomics for evolutionary studies and for transferring information from model species to related large-genome species has revolutionized molecular genetics and breeding strategies for improving those crops. Comparative sequence analysis methods can be used to cross-reference genes between species maps, enhance the resolution of comparative maps, study patterns of gene evolution, identify conserved regions of the genomes, and facilitate interspecies gene cloning. In this study, 5,780 Triticeae ESTs that have been physically mapped using wheat (Triticum aestivum L.) deletion lines and segregating populations were compared using NCBI BLASTN to the first draft of the public rice (Oryza sativa L.) genome sequence data from 3,280 ordered BAC/PAC clones. A rice genome view of the homoeologous wheat genome locations based on sequence analysis shows general similarity to the previously published comparative maps based on Southern analysis of RFLP. For most rice chromosomes there is a preponderance of wheat genes from one or two wheat chromosomes. The physical locations of non-conserved regions were not consistent across rice chromosomes. Some wheat ESTs with multiple wheat genome locations are associated with the non-conserved regions of similarity between rice and wheat. The inverse view, showing the relationship between the wheat deletion map and rice genomic sequence, revealed the breakdown of gene content and order at the resolution conferred by the physical chromosome deletions in the wheat genome. An average of 35% of the putative single copy genes that were mapped to the most conserved bins matched rice chromosomes other than the one that was most similar. This suggests that there has been an abundance of rearrangements, insertions, deletions, and duplications eroding the wheat-rice genome relationship that may complicate the use of rice as a model for cross-species transfer of information in non-conserved regions.