Consumption of Damson-hop Aphids (Phorodon humuli) by Larvae of Coccinella transversoguttata and Hippodamia convergens (Coleoptera: Coccinellidae)

Based on biomass, larvae of Hippodamia convergens Guerin consumed an average of 318 adult damson-hop aphids (Phorodon humuli (Schrank)), during their development at 20 C. Female larvae of Coccinella transversoguttata Falderman ate 413 adult P. humuli and males ate 357. This difference in the consumption of prey occurred only in the fourth stadium and was reflected in a corresponding size dimorphism between female and male larvae at pupation.     

Effects of plant spacing and interplanting with oilseed rape on colonisation of dwarf hops by the damson-hop aphid, Phorodon humuli

The pattern of colonisation of dwarf hops (Humulus lupulus) by damson-hop aphid (Phorodon humuli (Schrank)) migrating from Prunus spp. was investigated at six plant spacings and where some of the hops were replaced by oilseed rape (Brassica napus L.), a non-host of the aphid. The number of migrant aphids that accumulated on hop stems (bines) increased with increasing bine size and density. The numbers of aphids that colonised hops interplanted with oilseed rape reflected the density of the hop plants only and not the overall plant density. As the physical size of the wind shadow within which flying aphids can manoeuvre and land was unimportant unless provided by a host plant, the finding supports the theory that flying aphids respond to olfactory stimuli associated with their hosts. Variation in bine height (as a measure of plant size) explained 29–93% of the variance in aphid counts during the 3 years' study and bine density 1–14%. Standardising the data as the numbers of aphids per metre of bine and taking a square-root transformation of these standardised counts improved the precision of the analyses and, by stabilising variances, facilitated comparisons between years as growth became more vigorous as the plants matured. Each year, the rate of increase in numbers of aphids settling on plots of hops declined curvilinearly with increasing bine density. Maximum colonisation by P. humuli occurred at a bine density of five per metre row, a density similar to that used commercially by growers of dwarf hops.     

Electrical penetration graphs of the damson-hop aphid, Phorodon humuli on resistant and susceptible hops (Humulus lupulus)

Electrical penetration graphs (EPG's) were used to locate resistance to Phorodon humuli (Schrank) (Homoptera, Aphididae) in hops (Humulus lupulus, Cannabinaceae). Aphids on those hops showing resistance had a much reduced E2 pattern (uptake of phloem). In addition, many aphids on the resistant plants spent time non-probing within two minutes of withdrawing from the phloem. This was not observed with aphids on susceptible hops. The results suggest that resistance is located in the phloem. The involvement of a mechanical factor such as the blocking of aphid stylets, the presence of antinutritional factors, or simply an inadequate supply of nutrients, are discussed as possible resistance mechanisms.     

Increased Parasitization of Aphids on Trap Plants Alongside Vials Releasing Synthetic Aphid Sex Pheromone and Effective Range of the Pheromone

Aphid-infested cereal trap plants were used to detect the effects of aphid sex pheromone components on aphid parasitoid activity in arable field margins. The presence of aphid sex pheromones significantly increased parasitization levels by aphid parasitoids on the plants. By placing plants alongside and at varying distances away from a pheromone-releasing vial, the technique was used to measure the distance over which a point source of aphid sex pheromone could increase parasitization levels. Pheromone-releasing vials significantly increased parasitization by the generalist parasitoid Praon volucre on plants adjacent to vials and on plants placed 20 cm away. When the distance between pheromone-releasing vials and aphid-infested plants was increased to 1 m, parasitization by P. volucre was increased only on the plant adjacent to the vial, whereas parasitization by the specialist parasitoid Aphidius rhopalosiphi was also increased on plants placed 1 m away. This indicates a possible difference between the parasitoids in their foraging behaviour in response to semiochemical cues during host selection. When the experiment was repeated with some trap plants placed 3 m away from the pheromone-releasing vial, parasitization was again concentrated on plants directly alongside the vial, but only P. volucre appeared to be active in the field at the time of this experiment, so the effect on A. rhopalosiphi could not be assessed. The results are encouraging for the prospects of using aphid sex pheromones to manipulate parasitoids in order to improve aphid population control.     

Transmission of PPV-M to Prunus persica by Brachycaudus schwartzi and Phorodon humuli (Hem., Aphididae)

Abstract:  Aphids are reported to be vectors of the most serious viral pathogen of the drupaceous species plum pox virus (PPV), but there is little direct experimental evidence of this. PPV (serotype M) is widespread in peach orchards even where there are severe control measures. Laboratory bioassays were conducted to study, under controlled conditions, the ability of Brachycaudus schwartzi (Börner) and Phorodon humuli (Schrank) to transmit PPV (serotype M). The results have shown that all the peach trees tested had evident symptoms of sharka and were positive to the RT-PCR analysis, confirming the ability of these two aphid species to transmit the virus.     

The Common Black Ant, Lasius niger (Hymenoptera: Formicidae), as a Vector of the Entomopathogen Lecanicillium longisporum to Rosy Apple Aphid, Dysaphis plantaginea (Homoptera: Aphididae)

The potential for common black ant Lasius niger workers to vector conidia of the entomopathogenic fungus Lecanicillium longisporum to colonies of the rosy apple aphid Dysaphis plantaginea was assessed in laboratory and field experiments. Scanning electron microscope studies showed that L. niger workers which were artificially contaminated with L. longisporum conidia, carried conidia primarily on their tarsae but also on antennae and mandibles. Neither L. niger workers nor larvae were susceptible to infection with L. longisporum. Workers which were artificially contaminated with conidia of L. longisporum initiated infection in colonies of healthy D. plantaginea on apple. Mortality due to L. longisporum was recorded as 68.3, 30.8 and 3.7% of aphids under laboratory, semi-field and field conditions, respectively.     

Pathogenicity of the Hyphomycete Fungi Verticillium lecanii and Metarhizium anisopliae to the Western Flower Thrips,Frankliniella occidentalis

The pathogenicity of several isolates of the hyphomycete fungi Verticillium lecanii and Metarhizium anisopliae to Frankliniella occidentalis was investigated. Treatment of adult thrips with M. anisopliae resulted in at least 94% mortality at 7 days post-inoculation. In contrast, V. lecanii isolates only gave mortalities of between 20 and 70%. Detailed studies were made on the most virulent isolate of M. anisopliae (275) to determine its efficacy at different doses and temperatures. At 23 C the LC was ca. 3 105 conidia ml-1 after 5 days and the LTs were 50 50 3 and 4.5 days at 10 7 and 106 conidia ml-1 respectively. Temperature influenced fungal virulence to adult thrips; the LT at 18 and 20 C was ca. 4 days and at 23 or 26 C it was ca. 3 days. 50 Larvae were less susceptible to infection than adult thrips (27% versus 100% mortality), presumably due to the inoculum being shed with the exuvium during ecdysis. Conidia of M. anisopliae isolate 275 germinated rapidly on the surfaces of larvae, pupae and adults, with most germlings producing appressoria within 24 h post-inoculation. Fungal elements were present in significant amounts in the body 3 days after treatment.     

Pathogenicity of the Fungus Verticillium lecanii to Aphids and Powdery Mildew

Aspects of the host specificity and pathogenicity of the hyphomycete, Verticillium lecanii , were investigated under laboratory conditions. DiVerences were observed in the pathogenicities of three strains of V. lecanii (DAOM 198499, DAOM 216596 and Vertalec) to the potato aphid Macrosiphum euphorbiae and Sphaerotheca fuliginea , the causal agent of cucumber powdery mildew. The estimated median lethal concentration required to achieve 50% mortality (LC ), 50 median lethal time leading to 50% mortality (LT ) and aphid net reproductive rate ( R ) 50 0 indicated that Vertalec and strain 198499 were more virulent to aphids than strain 216596. The estimated median colonization time for 50% of fungal colonies (CT ) showed that strain 50 198499 was the best antagonist of cucumber powdery mildew. Further comparison suggested that the mean pathogenicity of V. lecanii strain 198499 to cucumber powdery mildew was almost equivalent to that of Sporothrix flocculosa , a biological control agent of greenhouse fungal pathogens. These observations provide convincing experimental evidence that V. lecanii is biologically active against both arthropods and fungi. The potential of using V. lecanii strain 198499 in biological control is discussed.     

Pheromone trapping models for pest control: Effects of mating patterns and immigration

Several models are presented which examine pest population behaviour with the release of female sex pheromones for the attraction and annihilation of males. These models include male polygamy and female monogamy, various mating frequencies, delayed mating of females, immigration of one or all individual types, and differential survivorship of males and females. In all the models there are two steady states, a stable s.s. at the origin and an unstable s.s. in the positive domain for a given value of pheromone release rate. In all the models, control relies on the reduced ability of males to fertilize virgin females following trapping and male annihilation. As such, control is very sensitive to mating frequency, being very difficult when males mate frequently. Control is also very difficult with the immigration of even a moderate number of fertilized females. Control is much easier when mating is delayed, especially if survivorship is low, or with density dependent population regulation.     

Selection of Verticillium lecanii Isolates with High Potential for Biocontrol of Cucumber Powdery Mildew by Means of Components Analysis at Different Humidity Regimes

To identify characteristics for the selection of Verticillium lecanii isolates with high potential for biocontrol of Sphaerotheca fuliginea under glasshouse conditions, an exploratory study was performed on the effect of water limitation on the development of 14 isolates. The conidial germination, growth and sporulation of isolates of V. lecanii were studied in a tritrophic system on cucumber leaves and in a ditrophic system in Petri dishes. Their mycoparasitic ability was studied in S. fuliginea and Cladosporium cladosporioides . All characteristics were clearly affected by humidity. Four isolates showed good biocontrol potential. The performance of isolates on agar had less predictive value than on powdery mildew. The germination of isolates of V. lecanii was lower and the mycelial growth faster on agar than on mildewed leaves under corresponding humidity conditions. The results suggest that conditions in the phyllosphere differed from the set humidity in the surrounding air. A correlation was found between the lysis of C. cladosporioides growing in dual culture on agar with isolates of V. lecanii and the parasitism of powdery mildew on detached, rooted leaves. C. cladosporioides might offer a suitable substrate for testing isolates of V. lecanii for mycoparasitic potential under various environmental conditions. Conidial germination, growth and sporulation had limited predictive value.     

Compatibility of Phloxine B, an Insecticidal Photoactive Dye, with Selected Biocontrol Fungi

The incorporation of the photoactive red food dye, phloxine B, into integrated pest management strategies has been suggested for the control of various insect pests. Inclusion of such a chemical pest control agent may interfere with a microbial pest control agent. Thus, representative fungi with potential for biocontrol were tested for their responses to phloxine B (0.01%) at different temperatures. Growth rates at selected temperatures were inhibited by dye in the presence of light for Coniothyrium minitans and Verticillium lecanii. Trichoderma virens growth was inhibited in light and dark. At optimal growth temperatures, Beauveria bassiana growth was inhibited only by the photoactivated dye species, while growth of Stilbella erythrocephala was not affected by the dye even in the presence of light.     

Identification of Functionally Important Residues of the Silkmoth Pheromone Biosynthesis-activating Neuropeptide Receptor,an Insect Ortholog of the Vertebrate Neuromedin U Receptor

The biosynthesis of sex pheromone components in many lepidopteran insects is regulated by the interaction between pheromone biosynthesis-activating neuropeptide (PBAN) and the PBAN receptor (PBANR), a class A G-protein-coupled receptor. To identify functionally important amino acid residues in the silkmoth PBANR, a series of 27 alanine substitutions was generated using a PBANR chimera C-terminally fused with enhanced GFP. The PBANR mutants were expressed in Sf9 insect cells, and their ability to bind and be activated by a core PBAN fragment (C10PBAN^R2K) was monitored. Among the 27 mutants, 23 localized to the cell surface of transfected Sf9 cells, whereas the other four remained intracellular. Reduced binding relative to wild type was observed with 17 mutants, and decreased Ca²⁺ mobilization responses were observed with 12 mutants. Ala substitution of Glu-95, Glu-120, Asn-124, Val-195, Phe-276, Trp-280, Phe-283, Arg-287, Tyr-307, Thr-311, and Phe-319 affected both binding and Ca²⁺ mobilization. The most pronounced effects were observed with the E120A mutation. A molecular model of PBANR indicated that the functionally important PBANR residues map to the 2nd, 3rd, 6th, and 7th transmembrane helices, implying that the same general region of class A G-protein-coupled receptors recognizes both peptidic and nonpeptidic ligands. Docking simulations suggest similar ligand-receptor recognition interactions for PBAN-PBANR and the orthologous vertebrate pair, neuromedin U (NMU) and NMU receptor (NMUR). The simulations highlight the importance of two glutamate residues, Glu-95 and Glu-120, in silkmoth PBANR and Glu-117 and Glu-142 in human NMUR1, in the recognition of the most functionally critical region of the ligands, the C-terminal residue and amide.     

Structure of the Mouse Sex Peptide Pheromone ESP1 Reveals a Molecular Basis for Specific Binding to the Class C G-protein-coupled Vomeronasal Receptor

Exocrine gland-secreting peptide 1 (ESP1) is a sex pheromone that is released in male mouse tear fluids and enhances female sexual receptive behavior. ESP1 is selectively recognized by a specific class C G-protein-coupled receptor (GPCR), V2Rp5, among the hundreds of receptors expressed in vomeronasal sensory neurons (VSNs). The specific sensing mechanism of the mammalian peptide pheromone by the class C GPCR remains to be elucidated. Here we identified the minimal functional region needed to retain VSN-stimulating activity in ESP1 and determined its three-dimensional structure, which adopts a helical fold stabilized by an intramolecular disulfide bridge with extensive charged patches. We then identified the amino acids involved in the activation of VSNs by a structure-based mutational analysis, revealing that the highly charged surface is crucial for the ESP1 activity. We also demonstrated that ESP1 specifically bound to an extracellular region of V2Rp5 by an in vitro pulldown assay. Based on homology modeling of V2Rp5 using the structure of the metabotropic glutamate receptor, we constructed a docking model of the ESP1-V2Rp5 complex in which the binding interface exhibited good electrostatic complementarity. These experimental results, supported by the molecular docking simulations, reveal that charge-charge interactions determine the specificity of ESP1 binding to V2Rp5 in the large extracellular region characteristic of class C GPCRs. The present study provides insights into the structural basis for the narrowly tuned sensing of mammalian peptide pheromones by class C GPCRs.     

Moderate drought influences the effect of arbuscular mycorrhizal fungi as biocontrol agents against Verticillium-induced wilt in pepper

Previous studies have shown that the arbuscular mycorrhizal fungus (AMF) Glomus deserticola (Trappe, Bloss and Menge) can diminish the negative effect of Verticillium dahliae Kleb. on pepper yield. On the other hand, it is known that AMF can be more beneficial for plant growth and physiology under dry conditions than when soil moisture is plentiful. Therefore, our objective was to assess if a moderate water deficit imposed on pepper plants before their inoculation with V. dahliae could improve the effectiveness of G. deserticola as biocontrol agent. In the present experiment, the delay in disease development in Verticillium-inoculated plants associated with AMF did not occur under well watered conditions. In addition, the establishment of mycorrhizal symbiosis and the development of structures by AMF were delayed when both symbiotic and pathogenic fungi infected the same root. Therefore, it is suggested that the equilibrium between pepper plant, G. deserticola and V. dahliae is so complex that small changes in competition between symbiotic and pathogenic fungi for host resources can modify the efficiency of AMF as a biocontrol agent. On the other hand, water deficit enhanced the deleterious effect of V. dahliae on fruit set and yield only when pepper plants were not associated with G. deserticola, which reinforces the idea that AMF may be more important for host plants subjected to stressful conditions. However, comparing well watered non-mycorrhizal and predroughted mycorrhizal plants, we found that moderate water deficit imposed before inoculation with V. dahliae did not improve the effectiveness of G. deserticola as a biocontrol agent.     

Colonization of the rhizosphere of crop plants by plant-beneficial pseudomonads

Abstract: Efficient colonization of the plant root is thought to be crucial for the plant-beneficial effect of particular Pseudomonas strains. Since root colonization is often the limiting step for successful plant growth stimulation, this process needs improvement. It is therefore important to acquire more information as regards (i) the conditions in the rhizosphere, and (ii) the bacterial traits that are involved in colonization. This review discusses some recent studies that focus on these two issues.     

Mechanisms of communication disruption by pheromone in the control of Lepidoptera: a review

ABSTRACT. Five possible mechanisms involved in the disruption of phero-mone-mediated behaviour in the control of lepidopterous pests are discussed. The various techniques used in the field, laboratory investigations of such mechanisms, and possible limitations in the application of such studies, are reviewed.     

Biosurfactants are involved in the biological control of Verticillium microsclerotia by Pseudomonas spp

AIMS: To examine the effect of previously described bacterial antagonists on the viability of Verticillium microsclerotia in vitro and to elucidate the possible modes of action of bacterial strains in the suppression of Verticillium microsclerotia viability. METHODS AND RESULTS: A microplate assay was developed to test the suppressive effect of well-defined Pseudomonas spp. on the viability of Verticillium microsclerotia in vitro. Experiments using phenazine- and biosurfactant-deficient mutants indicated that biosurfactants and phenazine-1-carboxylic acid play a role in the suppression of microsclerotia viability by Pseudomonas spp. In addition, microsclerotia colonization tests revealed that Pseudomonas spp. are able to colonize the surface of the microsclerotia, but not the inner matrix. Growth response curves showed that the population levels of Pseudomonas spp. increased when they were in the vicinity of Verticillium microsclerotia, indicating that Pseudomonas spp. may utilize nutrients from the microsclerotia for their growth. CONCLUSIONS: Pseudomonas spp. seem to be good candidates for Verticilllium microsclerotia biocontrol. Biosurfactant production is one of the main mechanisms involved in their mode of action. SIGNIFICANCE AND IMPACT OF THE STUDY: This line of work may contribute to a better understanding of biological control agents and their working mechanisms.