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Establishment and spread of invasive species can be facilitated by lack of natural enemies in the invaded area. Host-range evolution of natural enemies augments their ability to reduce the impact of the invader and could enhance their value for biological control. We assessed the potential of the Drosophila parasitoid, Leptopilina heterotoma (Hymenoptera: Figitidae), to exploit the invasive pest Drosophila suzukii by focusing on three performance indices: (i) attack rate; (ii) host killing, consisting of killing rate and lethal attack rate (killing efficiency); and (iii) successful offspring development (reproductive success). We found significant intraspecific variation in attack rate and killing rate and lethal attack rate among seven European populations, but offspring generally failed to successfully develop from the D. suzukii host. We crossed these European lines to create a genetically variable source population and performed a half-sib analysis to quantify genetic variation. Using a Bayesian animal model, we found that attack rate and killing rate had a heritability of h(2)=0.2, lethal attack rate h(2)=0.4, and offspring development h(2)=0. We then artificially selected wasps with the highest killing rate of D. suzukii for seven generations to test whether host-killing could be improved. There was a small and inconsistent response to selection in the three selection lines. Realized heritability (hr(2)) after four generations of selection was 0.17 but near zero after seven generations of selection. The genetic response might have been masked by an increased D. suzukii fitness resulting from adaptation to laboratory conditions. Our study reveals that native, European, L. heterotoma can attack the invasive pest, D. suzukii and significantly reduce fly survival and that different steps of the parasitization process need to be considered in the evolution of host-range. It highlights how evolutionary principles can be applied to optimize performance of native species for biological control.

Aim: The present study aimed to investigate the antifungal effects of fenhexamid, cyprodinil+fludioxanil, Bacillus subtilis QST 713, Trichoderma secondary metabolites (EGE-K-38 and EGE-K-71) on 6 isolates of necrotrophic fungi (Botrytis cinerea) under in- vitro conditions. Methodology: A dual culture technique was used to evaluate the effects of volatile compounds produced by Trichoderma in culture media against Botrytis cinerea isolates. For non-volatile and volatile metabolites, measurements for growth inhibition were made with the mixture of filter sterilized liquid medium of Trichoderma and potato dextrose agar. The fungicidal effects of these non-volatile metabolites and fungicides were first evaluated according to the effective concentration (EC50) data of growing mycelium inhibition of disease agents. Results: According to the results obtained, the drug/agent with the lowest fungicidal activity was determined as fenhexamide, which seems to have the lowest EC50 value of 0.05 ug m l-1. Both non-volatile metabolites and volatile metabolites of Trichoderma strains showed strong inhibition against B. cinerea isolates under experimental conditions. The highest growth inhibition percentage was determined with volatile metabolites of Trihoderma atroviride EGE-K-71 strain as 71.8%. Interpretation: The study suggest that filtrates that contain secondary metabolites and volatile compounds of Trichoderma strains produced in both liquid culture and dual culture conditions can be used as an effective fungal control agents against pre and postharvest contamination of grapes with Botrytis cinerea.

Insects have been on the menu of humans for centuries, but only recently we have begun to mass produce them for human food and animal feed. This introduction first paints a synopsis of mass cultured insects and their application. The new insect production industry raises many interesting fundamental and applied questions about insect biology and fitness. The second part of the introduction to this special issue addresses the 13 articles dealing with the improvement of mass-rearing efforts for a range of insects. The various studies focus on the effects of diet and microorganisms on relevant life-history traits and economic value of the insects. They reflect the current rapid developments in the insect production industry.

Area-wide integrated pest management strategies against tephritid fruit flies include the release of fruit fly parasitic wasps in the target area. Mass rearing of parasitic wasps is essential for the efficient application of biological control strategies. Enhancement of fruit fly host fitness through manipulation of their gut-associated symbionts might also enhance the fitness of the produced parasitic wasps and improve the parasitoid rearing system. In the current study, we added three gut bacterial isolates originating from Ceratitis capitata (Wiedemann) and four originating from Bactrocera oleae (Rossi) (both Diptera: Tephritidae) to the larval diet of C. capitata and used the bacteria-fed larvae as hosts for the development of the parasitic wasp Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae). We evaluated the effect of the bacteria on wasp life-history traits and assessed their potential use for the improvement of D. longicaudata rearing. Enterobactersp. AA26 increased fecundity and parasitism rate and accelerated parasitoid emergence. Providencia sp. AA31 led to faster emergence of both male and female parasitoids, whereas Providencia sp. 22 increased the production of female progeny. Bacillus sp. 139 increased parasitoid fecundity, parasitism rate, and production of female progeny. Serratia sp. 49 accelerated parasitoid emergence for both males and females and increased production of female progeny. Klebsiella oxytoca delayed parasitoid emergence and Enterobacter sp. 23 decreased parasitoid fecundity and parasitism rate. Our findings demonstrate a wide range of effects of fruit fly gut symbionts on parasitoid production and reveal a great potential of bacteria use towards enhancement of parasitic wasp rearing.

Botryosphaeria dothidea causes apple ring rot, which is among the most prevalent postharvest diseases of apples and causes significant economic loss during storage. In this study, we investigated the biocontrol activity and possible mechanism of Bacillus velezensis strain P2-1 isolated from apple branches against B. dothidea in postharvest apple fruit. The results showed strain P2-1, one of the 80 different endophytic bacterial strains from apple branches, exhibited strong inhibitory effects against B. dothidea growth and resulted in hyphal deformity. B. velezensis P2-1 treatment significantly reduced the ring rot caused by B. dothidea. Additionally, the supernatant of strain P2-1 exhibited antifungal activity against B. dothidea. Re-isolation assay indicated the capability of strain P2-1 to colonize and survive in apple fruit. PCR and qRT-PCR assays revealed that strain P2-1 harbored the gene clusters required for biosynthesis of antifungal lipopeptides and polyketides. Strain P2-1 treatment significantly enhanced the expression levels of pathogenesis-related genes (MdPR1 and MdPR5) but did not significantly affect apple fruit qualities (measured in fruit firmness, titratable acid, ascorbic acid, and soluble sugar). Thus, our results suggest that B. velezensis strain P2-1 is a biocontrol agent against B. dothidea-induced apple postharvest decay. It acts partially by inhibiting mycelial growth of B. dothidea, secreting antifungal substances, and inducing apple defense responses.

Disease suppressive soils are important for managing soil-borne diseases that cannot be controlled with chemicals. One such disease is the potato common scab caused by Streptomyces species. Suppressiveness against common scab can develop spontaneously in fields where potato is grown for years without interruption, and this has been attributed to non-pathogenic Streptomyces strains. Streptomyces spp. have been used as inoculants in biological control, but their long-term effects have gained less attention. In our previous studies, a nonpathogenic Streptomyces strain (Str272) isolated from a potato common scab lesion suppressed common scab in field trials lasting over 5 years. In this study, bacterial communities in the tuberosphere i.e. in the soil adjacent to potato tubers, were analysed by next generation sequencing (NGS). The aim was to compare bacterial communities in untreated control plots to those in which seed tubers were treated with Str272 in one or several growing seasons. Str272 applications increased soil bacterial diversity and affected the bacterial composition in the potato tuberosphere. The most pronounced differences were observed between the untreated control and the treatments in which the antagonist had been applied in three or four consecutive years. The differences remained similar until the following growing season. Bacterial composition after repeated antagonist applications was associated with lower common scab severity. The antagonist applications had no or only slight effect on the number or abundance of OTUs belonging to Actinobacteria or Streptomyces, and no differences in quantities of pathogenic Streptomyces populations were detected by qPCR. This indicates that suppression of common scab by Str272 may not be based on direct effect on the common scab pathogens but is more likely to be associated with the alterations of the soil bacterial community. The most abundant bacteria phyla in the potato tuberosphere were Actinobacteria, Proteobacteria and Acidobacteria. However, the OTUs responding greatest to the antagonist treatments belonged to Bacterioidetes and Gemmatimonadetes. Results indicate that repeated applications of Str272 can change the bacterial community in the potato tuberosphere and lead to development of soil that is suppressive against potato common scab for several growing seasons after the last application.

Biochar has been reported to play a positive role in disease suppression against airborne pathogens in plants. The mechanisms behind this positive trait are not well-understood. In this study, we hypothesized that the attraction of plant growth-promoting rhizobacteria (PGPR) or fungi (PGPF) underlies the mechanism of biochar in plant protection. The attraction of PGPR and PGPF may either activate the innate immune system of plants or help the plants with nutrient uptake. We studied the effect of biochar in peat substrate (PS) on the susceptibility of strawberry, both on leaves and fruits, against the airborne fungal pathogen Botrytis cinerea. Biochar had a positive impact on the resistance of strawberry fruits but not the plant leaves. On leaves, the infection was more severe compared with plants without biochar in the PS. The different effects on fruits and plant leaves may indicate a trade-off between plant parts. Future studies should focus on monitoring gene expression and metabolites of strawberry fruits to investigate this potential trade-off effect. A change in the microbial community in the rhizosphere was also observed, with increased fungal diversity and higher abundances of amplicon sequence variants classified into Granulicella, Mucilaginibacter, and Byssochlamys surrounding the plant root, where the latter two were reported as biocontrol agents. The change in the microbial community was not correlated with a change in nutrient uptake by the plant in either the leaves or the fruits. A decrease in the defense gene expression in the leaves was observed. In conclusion, the decreased infection of B. cinerea in strawberry fruits mediated by the addition of biochar in the PS is most likely regulated by the changes in the microbial community.

Harmonia axyridis is an invasive alien ladybird in North America and Europe. Studies show that multiple natural enemies are using Ha. axyridis as a new host. However, thus far, no research has been undertaken to study the effects of simultaneous infections of multiple natural enemies on Ha. axyridis. We hypothesized that high thallus densities of the ectoparasitic fungus Hesperomyces virescens on a ladybird weaken the host’s defenses, thereby making it more susceptible to infection by other natural enemies. We examined mortality of the North American-native Olla v-nigrum and Ha. axyridis co-infected with He. virescens and an entomopathogenic fungus—either Beauveria bassiana or Metarhizium anisopliae. Laboratory assays revealed that He. virescens-infected O. v-nigrum individuals are more susceptible to entomopathogenic fungi, but Ha. axyridis does not suffer the same effects. This is in line with the enemy release hypothesis, which predicts that invasive alien species in new geographic areas experience reduced regulatory effects from natural enemies compared to native species. Considering our results, we can ask how He. virescens affects survival when confronted by other pathogens that previously had little impact on Ha. axyridis.

Sphecophaga vesparum often parasitizes nests of vespid wasps such as Vespula vulgaris and Vespula germanica. Inside the colonies, the ectoparasitic larvae feed on the immature forms of the wasps. There are two adult forms of S. vesparum. The large, winged adults emerge from either rigid yellow cocoons or the orange cocoons used for overwintering. The small, brachypterous females emerge from soft, white cocoons. The species is facultative deuterotokous, producing mostly parthenogenic females and infrequently producing males. Here, we describe the production of chemical compounds related to the different developmental forms of the parasitoid S. vesparum (larvae, pupae and adults). We also compare the chemical profiles of the parasitoid wasp adults to those of their two main host species, Vespula vulgaris and Vespula germanica. The results show differences in hydrocarbon composition of larvae, pupae and adults of S. vesparum. Our results also suggest a partial mimicry of each of the two host species, mostly relating to linear alkanes present in both parasitoids and the host vespid wasp species. This matching is likely due to the recycling of the prey&rsquo, s hydrocarbons, as has been found in other species of parasitoids.

New genetic control methods for mosquitoes may reduce vector species without direct effects on other species or the physical environment common with insecticides or drainage. Effects on predators and competitors could, however, be a concern as Anopheles gambiae s.l. is preyed upon in all life stages. We overview the literature and assess the strength of the ecological interactions identified. Most predators identified consume many other insect species and there is no evidence that any species preys exclusively on any anopheline mosquito. There is one predatory species with a specialisation on blood‐fed mosquitoes including An. gambiae s.l.. Evarcha culicivora is a jumping spider, known as the vampire spider, found around Lake Victoria. There is no evidence that these salticids require Anopheles mosquitoes and will readily consume blood‐fed Culex. Interspecific competition studies focus on other mosquitoes of larval habitats. Many of these take place in artificial cosms and give contrasting results to semi‐field studies. This may limit their extrapolation regarding the potential impact of reduced An. gambiae numbers. Previous mosquito control interventions are informative and identify competitive release and niche opportunism; so while the identity and relative abundance of the species present may change, the biomass available to predators may not.

Plant root secreted compounds alter the gene expression of associated microorganisms by acting as signal molecules that either stimulate or repel the interaction with beneficial or harmful species, respectively. However, it is still unclear whether two distinct groups of beneficial bacteria, non-plant-associated (soil) strains and plant-associated (endophytic) strains, respond uniformly or variably to the exposure with root exudates. Therefore, Bacillus mycoides, a potential biocontrol agent and plant growth-promoting bacterium, was isolated from the endosphere of potatoes and from soil of the same geographical region. Confocal fluorescence microscopy of plants inoculated with GFP-tagged B. mycoides strains showed that the endosphere isolate EC18 had a stronger plant colonization ability and competed more successfully for the colonization sites than the soil isolate SB8. To dissect these phenotypic differences, the genomes of the two strains were sequenced and the transcriptome response to potato root exudates was compared. The global transcriptome profiles evidenced that the endophytic isolate responded more pronounced than the soil-derived isolate and a higher number of significant differentially expressed genes were detected. Both isolates responded with the alteration of expression of an overlapping set of genes, which had previously been reported to be involved in plant-microbe interactions; including organic substance metabolism, oxidative reduction, and transmembrane transport. Notably, several genes were specifically upregulated in the endosphere isolate EC18, while being oppositely downregulated in the soil isolate SB8. These genes mainly encoded membrane proteins, transcriptional regulators or were involved in amino acid metabolism and biosynthesis. By contrast, several genes upregulated in the soil isolate SB8 and downregulated in the endosphere isolate EC18 were related to sugar transport, which might coincide with the different nutrient availability in the two environments. Altogether, the presented transcriptome profiles provide highly improved insights into the life strategies of plant-associated endophytes and soil isolates of B. mycoides.

The entomopathogenic fungus Beauveria bassiana is able to colonize and establish a symbiotic relationship with plants as an endophyte, naturally as well as through artificial inoculation. This study aims at implementing a reliable and efficient method to establish endophytic B. bassiana in maize plants. The level of the endophytic establishment was evaluated and the survival and fecundity of the English grain aphid (Sitobion avenae) was investigated. Eight-days-old maize plants were sprayed with a fungal spore suspension, while control plants were treated only with a Tween solution. Two weeks after inoculation, seven clip cages per plant, each containing an adult aphid, were attached to different leaves and left for seven days. All inoculated plants contained endophytic B. bassiana as confirmed by plating surface disinfected leaf pieces on selective plates. None of the control plants showed growth of B. bassiana. The highest colonization levels were 61% in older inoculated leaves and 19% in younger non-inoculated leaves. The survival of S. avenae on inoculated plants was reduced up to 49% as compared to control plants. Dead aphids did not show any signs of mycosis. The number of nymphs produced by a single aphid was reduced up to 70% and 74% on inoculated and non-inoculated leaves, respectively. Presence of endophytic B. bassiana in non-inoculated younger leaves implies the movement of the endophyte inside plants. The negative effect on aphid fecundity appears to be systemic as implied by the almost equal reduction of mean numbers of nymphs produced on older and younger leaves on the inoculated plants, irrespective of the level of colonization by endophytic B. bassiana.

Globally, Anastatus species (Hymenoptera: Eupelmidae) are associated with the invasive agricultural pest Halyomorpha halys (Stal) (Hemiptera: Pentatomidae). In Europe, the polyphagous Anastatus bifasciatus (Geoffroy) is the most prevalent native egg parasitoid on H. halys eggs and is currently being tested as a candidate for augmentative biological control. Anastatus bifasciatus frequently displays behavior without oviposition, and induces additional host mortality through oviposition damage and host feeding that is not measured with offspring emergence. This exacerbates accurate assessment of parasitism and host impact, which is crucial for efficacy evaluation as well as for pre- and post-release risk assessment. To address this, a general Anastatus primer set amplifying a 318-bp fragment within the barcoding region of the cytochrome oxidase I (COI) gene was developed. When challenged with DNA of three Anastatus species -A. bifasciatus, Anastatus japonicus Ashmead, and Anastatus sp.-, five scelionid parasitoid species that might be encountered in the same host environments and 11 pentatomid host species, only Anastatus DNA was successfully amplified. When applied to eggs of the target host, H. halys, and an exemplary non-target host, Dendrolimus pini L. (Lepidoptera: Lasiocampidae), subjected to host feeding, no Anastatus amplicons were produced. Eggs of the two host species containing A. bifasciatus parasitoid stages, from 1-h-old eggs to pupae, and emerged eggs yielded Anastatus fragments. Confirmation of parasitoid presence with dissections and subsequent PCRs with the developed primer pair resulted in 95% success for 1-h-old parasitoid eggs. For both host species, field-exposed sentinel emerged eggs stored dry for 6 months, 100% of the specimens produced Anastatus amplicons. This DNA-based screening method can be used in combination with conventional methods to better interpret host-parasitoid and parasitoid-parasitoid interactions. It will help address ecological questions related to an environmentally friendly approach for the control of H. halys in invaded areas.

In hymenopterans, males are normally haploid (1n) and females diploid (2n), but individuals with divergent ploidy levels are frequently found. In species with ‘complementary sex determination’ (CSD), increasing numbers of diploid males that are often infertile or unviable arise from inbreeding, presenting a major impediment to biocontrol breeding. Non‐CSD species, which are common in some parasitoid wasp taxa, do not produce polyploids through inbreeding. Nevertheless, polyploidy also occurs in non‐CSD Hymenoptera. As a first survey on the impacts of inbreeding and polyploidy of non‐CSD species, we investigate life‐history traits of a long‐term laboratory line of the parasitoid Nasonia vitripennis (Walker) (Hymenoptera: Pteromalidae) (‘Whiting polyploid line’) in which polyploids of both sexes (diploid males, triploid females) are viable and fertile. Diploid males produce diploid sperm and virgin triploid females produce haploid and diploid eggs. We found that diploid males did not differ from haploid males with respect to body size, progeny size, mate competition, or lifespan. When diploid males were mated to many females (without accounting for mating order), the females produced a relatively high proportion of male offspring, possibly indicating that these males produce less sperm and/or have reduced sperm functionality. In triploid females, parasitization rate and fecundity were reduced and body size was slightly increased, but there was no effect on lifespan. After one generation of outbreeding, lifespan as well as parasitization rate were increased, and a body size difference was no longer apparent. This suggests that outbreeding has an effect on traits observed in an inbred polyploidy background. Overall, these results indicate some phenotypic detriments of non‐CSD polyploids that must be taken into account in breeding.

Artificial sugar solutions can be used to disrupt the mutualism between aphids and ants. This provides natural enemies with increased access to aphid populations, thus enhancing biocontrol of hemipteran pests. However, research suggests that artificial solutions should be tailored to specific species in order to maximize the desired result. In this study, we test the effect of an alkaloid, caffeine, and a non-protein amino acid, γ-aminobutyric acid (GABA), on preference for sucrose solutions in two ant species, Lasius niger and Oecophylla smaragdina. These secondary metabolites have been shown to have attractive capabilities in other species of insects, who favor nectars from plants containing these compounds. In our first experiment, both species significantly preferred solutions containing GABA over sucrose-only controls, albeit L. niger favored higher concentrations of the amino acid. Caffeine did not significantly increase preference for sugar solutions in either of the tested species, and ants significantly dispreferred the highest concentration of caffeine offered. When the two metabolites were tested simultaneously against sucrose only, L. niger fed equally from all solutions, while O. smaragdina dispreferred caffeine. Thus, while GABA seems to be a promising ant attractant, caffeine is not an adequate choice, at least at the concentrations tested in this experiment.

Agricultural practices often involve tank-mixing and co-application of insecticides with fungicides to control crop pests. However, natural methods relying on biological control agents such as hymenopteran parasitoids have been shown to be highly effective in suppressing crop pest populations. The current body of insecticide risk assessment data accounting for fungicide co-application is very small, the present study being the first to examine this in a parasitoid wasp. Through low-dose exposure to dry residues of the neonicotinoid insecticide thiacloprid, we examined its mortal and knockdown effect on Aphelinus abdominalis when co-applied with increasing doses of the fungicide tebuconazole. Both of these acute effects of thiacloprid were synergised (toxicity increased to a greater-than-additive effect) by tebuconazole, resulting in significant mortality from low-dose co-applications of tebuconazole, and significant knockdown even without co-applied tebuconazole, the effect increasing as tebuconazole concentration increased. We show the highly toxic effect that a low dose of thiacloprid imposes on A. abdominalis populations, and a synergistic toxicity when co-applied with low doses of tebuconazole. Our work suggests a need for updating pesticide risk assessment methods, accounting for pesticide mixtures, in order to make these risk assessments more field relevant.

Fusarium graminearum can cause Giberella Ear Rot (GER) and seedling blight in maize, resulting in major yield losses. Besides GER, the infected grains are consequently contaminated with multiple mycotoxins of F. graminearum. Zearalenone and trichothecenes, such as deoxynivalenol and its acetylated forms, are among the major mycotoxins associated with F. graminearum infection in maize. In the current work, we explored the effect of the endophytic fungal genera of Epicoccum and Sordaria, to control F. graminearum infection in comparative trials with Piriformospora spp., an elusive endophytic genus. Furthermore, we investigated the effect of these endophytes on zearalenone, deoxynivalenol, and 15-acetyldeoxynivalenol levels using in vitro and in planta assays. As plants are endowed with several detoxification mechanisms comprising e.g., glucosylation of trichothecenes, the effect of the isolated fungal endophytes on the deoxynivalenol-3-glucoside level was also assessed. In general, results showed a considerable variability in the antifungal activity, both among species and among isolates within one species. Additionally, the effect on mycotoxin levels was variable, and not necessarily related to the antifungal activity except for zearalenone levels which were consistently reduced by the endophytes. These results highlight the great potential of certain endophytic fungal strains as new biocontrol agents in agricultural science.

Plant microbiome plays an important role in maintaining the host fitness. Despite a significant progress in our understanding of the plant microbiome achieved in the recent years, very little is known about the effect of plant pathogens on composition of microbial communities associated with trees. In this study, we analysed the mycobiome of different anatomic parts of asymptomatic and symptomatic Norway spruce trees naturally infected by Heterobasidion spp. We also investigated the primary impact of the disease on the fungal communities, which are associated with Norway spruce trees. Our results demonstrate that the structure of fungal communities residing in the wood differed significantly among symptomatic and asymptomatic Heterobasidion-infected trees. However, no significant differences were found in the other anatomic regions of the trees. The results also show that not only each of individual tree tissues (wood, bark, needles and roots) harbours a unique fungal community, but also that symptomatic trees were more susceptible to co-infection by other wood-degrading fungi compared to the asymptomatic ones.

Peat based growing media are not ecologically sustainable and often fail to support biological control. Miscanthus straw was (1) tested to partially replace peat; and (2) pre-colonized with a Trichoderma strain to increase the biological control capacity of the growing media. In two strawberry pot trials (denoted as experiment I & II), extruded and non-extruded miscanthus straw, with or without pre-colonization with T. harzianum T22, was used to partially (20% v/v) replace peat. We tested the performance of each mixture by monitoring strawberry plant development, nutrient content in the leaves and growing media, sensitivity of the fruit to the fungal pathogen Botrytis cinerea, rhizosphere community and strawberry defense responses. N immobilization by miscanthus straw reduced strawberry growth and yield in experiment II but not in I. The pre-colonization of the straw with Trichoderma increased the post-harvest disease suppressiveness against B. cinerea and changed the rhizosphere fungal microbiome in both experiments. In addition, defense-related genes were induced in experiment II. The use of miscanthus straw in growing media will reduce the demand for peat and close resource loops. Successful pre-colonization of this straw with biological control fungi will optimize crop cultivation, requiring fewer pesticide applications, which will benefit the environment and human health.

There is considerable evidence that both plant diversity and plant identity can influence the level of predation and predator abundance aboveground. However, how the level of predation in the soil and the abundance of predatory soil fauna are related to plant diversity and identity remains largely unknown. In a biodiversity field experiment, we examined the effects of plant diversity and identity on the infectivity of entomopathogenic nematodes (EPNs, Heterorhabditis and Steinernema spp.), which prey on soil arthropods, and abundance of carnivorous non-EPNs, which are predators of other nematode groups. To obtain a comprehensive view of the potential prey/food availability, we also quantified the abundance of soil insects and nonpredatory nematodes and the root biomass in the experimental plots. We used structural equation modeling (SEM) to investigate possible pathways by which plant diversity and identity may affect EPN infectivity and the abundance of carnivorous non-EPNs. Heterorhabditis spp. infectivity and the abundance of carnivorous non-EPNs were not directly related to plant diversity or the proportion of legumes, grasses and forbs in the plant community. However, Steinernema spp. infectivity was higher in monocultures of Festuca rubra and Trifolium pratense than in monocultures of the other six plant species. SEM revealed that legumes positively affected Steinernema infectivity, whereas plant diversity indirectly affected the infectivity of Heterorhabditis EPNs via effects on the abundance of soil insects. The abundance of prey (soil insects and root-feeding, bacterivorous, and fungivorous nematodes) increased with higher plant diversity. The abundance of prey nematodes was also positively affected by legumes. These plant community effects could not be explained by changes in root biomass. Our results show that plant diversity and identity effects on belowground biota (particularly soil nematode community) can differ between organisms that belong to the same feeding guild and that generalizations about plant diversity effects on soil organisms should be made with great caution.

[EN] The production of new generation of wood preservatives (without addition of a co-biocide) in combination with an exchange of wood poles on identical sites with high fungal inoculum, has resulted in an increase of premature failures of wood utility poles in the last decades. Wood destroying basidiomycetes inhabiting sites where poles have been installed, have developed resistance against wood preservatives. The objective of the in vitro studies was to identify a Trichoderma spp. with a highly antagonistic potential against wood destroying basidiomycetes that is capable of colonizing Cu-rich environments. For this purpose, the activity of five Trichoderma spp. on Cu-rich medium was evaluated according to its growth and sporulation rates. The influence of the selected Trichoderma spp. on wood colonization and degradation by five wood destroying basidiomycetes was quantitatively analyzed by means of dry weight loss of wood specimens. Furthermore, the preventative effect of the selected Trichoderma spp. in combination with four Cu-based preservatives was also examined by mass loss and histological changes in the wood specimens. Trichoderma harzianum (T-720) was considered the biocontrol agent with higher antagonistic potential to colonize Cu-rich environments (up to 0.1% CuSO4 amended medium). T. harzianum demonstrated significant preventative effect on wood specimens against four wood destroying basidiomycetes. The combined effect of T. harzianum and Cu-based wood preservatives demonstrated that after 9 months incubation with two wood destroying basidiomycetes, wood specimens treated with 3.8 kg m-3 copper-chromium had weight losses between 55±65%, whereas containers previously treated with T. harzianum had significantly lower weight losses (0±25%). Histological studies on one of the wood destroying basidiomycetes revealed typical decomposition of wood cells by brown-rot fungi in Cu-impregnated samples, that were notably absent in wood specimens previously exposed to T. harzianum. It is concluded that carefully selected Trichoderma isolates can be used for integrated wood protection against a range of wood destroying basidiomycetes and may have potential for integrated wood protection in the field., The authors are pleased to acknowledge the financial support by the Swiss CTI (Project No. 17001.1 PFLS-LS).

Particular groups of plant-beneficial fluorescent pseudomonads are not only root colonizers that provide plant disease suppression, but in addition are able to infect and kill insect larvae. The mechanisms by which the bacteria manage to infest this alternative host, to overcome its immune system, and to ultimately kill the insect are still largely unknown. However, the investigation of the few virulence factors discovered so far, points to a highly multifactorial nature of insecticidal activity. Antimicrobial compounds produced by fluorescent pseudomonads are effective weapons against a vast diversity of organisms such as fungi, oomycetes, nematodes, and protozoa. Here, we investigated whether these compounds also contribute to insecticidal activity. We tested mutants of the highly insecticidal strains Pseudomonas protegens CHA0, Pseudomonas chlororaphis PCL1391, and Pseudomonas sp. CMR12a, defective for individual or multiple antimicrobial compounds, for injectable and oral activity against lepidopteran insect larvae. Moreover, we studied expression of biosynthesis genes for these antimicrobial compounds for the first time in insects. Our survey revealed that hydrogen cyanide and different types of cyclic lipopeptides contribute to insecticidal activity. Hydrogen cyanide was essential to full virulence of CHA0 and PCL1391 directly injected into the hemolymph. The cyclic lipopeptide orfamide produced by CHA0 and CMR12a was mainly important in oral infections. Mutants of CMR12a and PCL1391 impaired in the production of the cyclic lipopeptides sessilin and clp1391, respectively, showed reduced virulence in injection and feeding experiments. Although virulence of mutants lacking one or several of the other antimicrobial compounds, i.e., 2,4-diacetylphloroglucinol, phenazines, pyrrolnitrin, or pyoluteorin, was not reduced, these metabolites might still play a role in an insect background since all investigated biosynthetic genes for antimicrobial compounds of strain CHA0 were expressed at some point during insect infection. In summary, our study identified new factors contributing to insecticidal activity and extends the diverse functions of antimicrobial compounds produced by fluorescent pseudomonads from the plant environment to the insect host., Frontiers in Microbiology, 8, ISSN:1664-302X

Chitin and its derivatives are natural biopolymers that are often used as compounds for the control of soilborne plant pathogens. In spite of recent advances in agricultural practices involving chitin amendments, the microbial communities in chitin-amended soils remain poorly known. The objectives of this study were (1) to investigate the bacterial diversity and abundance in an agricultural soil supplemented with chitin that turned disease-suppressive and (2) to assess the emergence of chitinolytic bacteria under conditions of raised soil pH. Amplicon pyrosequencing of soil-extracted DNA based on the 168 rRNA genes was used to characterize the structures of bacterial communities in soil, chitin-amended or not, with native versus raised pH (5.7 vs 8.7), in microcosms and the field. As a result of chitin addition, changes in the relative abundances of Actinobacteria, Proteobacteria and Bacteroidetes were observed in the field soil. A large and significant increase of the relative abundance of Oxalobacteraceae (Betaproteobacteria, Burkholderiales) was found. Within the Oxalobacteraceae, the genera Duganella and Massilia showed large increases. Moreover, responses of the Alpha- and Gammaproteobacteria appeared shortly after the alteration of the soil pH in the microcosms. A significant decrease in the abundance of Actinobacteria was observed in the chitin-amended field soil and in the microcosm at high pH. Overall, the bacterial abundance in soil tended to decrease with the addition of chitin. Two groups, Actinobacteria and Oxalobacteraceae, were found to be most responsive to the amendment. These results enhance the understanding of responses to chitin and possible interactions within bacterial communities in soil that can be correlated to soil disease suppressiveness. (C) 2014 Elsevier Ltd. All rights reserved.

There is an urgent need to test and develop sustainable methods for management of soil pathogens, such as the root-lesion nematode Pratylenchus penetrans and the soil fungus Verticillium dahliae. Ultimately this should be investigated with a multidisciplinary approach, with long-term measurements of biological and chemical parameters and their final impact on crop yield under field conditions. The present study focusses on eight soil health treatments (compost, chitin, marigold, grass–clover, biofumigation, anearobic soil disinfestation, a physical control method and a combination of marigold, compost and chitin) and two control treatments (a chemical control with 300 L/ha Metam sodium and un untreated control). These 10 treatments were studied for their effects on soil chemical quality, soil pathogens and their impact on the yield of potato, lily and carrots during 6 years. The present study did demonstrate that in comparison to chemical control, additions of chitin, anaerobic soil disinfestation and marigold are already excellent alternatives for the control of plant-parasitic nematodes and V. dahliae. We also demonstrated that grass–clover, biofumigation, Cultivit and compost are not effective alternatives for chemical control yet and further development is needed. All treatments caused a yield increase in comparison with the control. The biggest increases of more than 60% were found for the treatments with chitin. Furthermore it was demonstrated that these yield increases were probably less influenced by changes in chemical soil properties, but the consequence of changes in the soil biota, in this case especially the effective control of P. penetrans and V. dahliae. Furthermore it has been demonstrated that most of these soil health treatments could already be implemented in an arable crop rotation and probably adapted for many areas of the world where other alternatives, such as solarisation or soil flooding, are not feasible or too risky.

The bronze bug, Thaumastocoris peregrinus Carpintero et Dellape (Heteroptera Thaumastocoridae), is one of the most important emerging pests of Eucalyptus LHeritier plantations worldwide. In the development of strategies to control this pest, establishing effective rearing protocols is fundamental to future research programs. We assessed life-history parameters of the bronze bug in a newly designed mass rearing strategy. Separated units were set up to contain different developmental stages. Egg production by females reared on commonly found Eucalyptus grandis Hill ex Maiden and Eucalyptus tereticornis Smith was evaluated in order to determine which plant species to use in rearing. Females laid more eggs on E. tereticornis than on E. grandis, so the former species was chosen for the rearing. A cohort of 207 eggs was followed in Petri dishes until the last individual died or reached the adult stage. We followed egg production by 15 adult couples from the original cohort. Preparation of 150-200 dishes with hatching eggs per week allows for an average production of 7,500 eggs per week. Under our rearing conditions, eggs started hatching on day six, and the first adults were obtained 23 days after oviposition. Almost half of the eggs did not hatch, and the highest nymphal mortality was recorded in the second instar, while the lowest mortality occurred in the last instar. We discuss the relevance of this mass rearing strategy, both within the context of basic behavioural studies of T. peregrinus, and as a tool for the mass rearing of the biological control agent, Cleruchoides noackae Lin et Huber.

A long-term experiment on the effect of chitin addition to soil on the suppression of soilborne pathogens was set up and monitored for 8 years in an experimental field, Vredepeel, The Netherlands. Chitinous matter obtained from shrimps was added to soil top layers on two different occasions, and the suppressiveness of soil toward Verticillium dahliae , as well as plant-pathogenic nematodes, was assessed, in addition to analyses of the abundances and community structures of members of the soil microbiota. The data revealed that chitin amendment had raised the suppressiveness of soil, in particular toward Verticillium dahliae , 9 months after the (second) treatment, extending to 2 years following treatment. Moreover, major effects of the added chitin on the soil microbial communities were detected. First, shifts in both the abundances and structures of the chitin-treated soil microbial communities, both of total soil bacteria and fungi, were found. In addition, the abundances and structures of soil actinobacteria and the Oxalobacteraceae were affected by chitin. At the functional gene level, the abundance of specific (family-18 glycoside hydrolase) chitinase genes carried by the soil bacteria also revealed upshifts as a result of the added chitin. The effects of chitin noted for the Oxalobacteraceae were specifically related to significant upshifts in the abundances of the species Duganella violaceinigra and Massilia plicata . These effects of chitin persisted over the time of the experiment.

Significant stimulation of germination of tomatoes (Lycopersicon esculentum Mill. cv Odessa plum) with poor initial germination capacity was achieved by soaking their seeds during 6 h in suspensions of nine out of ten Lactobacillus plantarum strains tested. The increase in percentage of germination ranged from 15.1 to 7.6 %. Lengths of shoots, main and lateral roots and development of root hairs increased after inoculation of seeds with studied L. plantarum strains. The treatments of seeds with L. plantarum expanded shoot lengths by 16.4–18.2 %. Stimulation of main root lengths in the tested tomato seedlings varied from 10.5 to 31.4 %, and the stimulation of lateral roots from 28.6 to 32.2 %. Lactobacilli from different strains exert different effects on the seedlings: for instance, inoculation with certain strains resulted in the stimulation of the main root growth and the inoculation with another strain stimulated shoot growth or growth of the lateral roots. It is the first report about plant-stimulating activities of L. plantarum from different ecological niches (plant surfaces, surfaces of edible mushrooms, dairy products) in the absence of nutritional compounds.

BACKGROUND: Most insect-resistant transgenic crops employ toxins to control pests. A novel approach is to enhance the effectiveness of natural enemies by genetic engineering of the biosynthesis of volatile organic compounds (VOCs). Before the commercialisation of such transgenic plants can be pursued, detailed fundamental studies of their effects on herbivores and their natural enemies are necessary. The linalool/nerolidol synthase gene FaNES1 was constitutively expressed from strawberry in three Arabidopsis thaliana accessions, and the behaviour of the aphid Brevicoryne brassicae L., the parasitoid Diaeretiella rapae McIntosh and the predator Episyrphus balteatus de Geer was studied. RESULTS: Transgenic FaNES1-expressing plants emitted (E)-nerolidol and larger amounts of (E)-DMNT and linalool. Brevicoryne brassicae was repelled by the transgenic lines of two of the accessions, whereas its performance was not affected. Diaeretiella rapae preferred aphid-infested transgenic plants over aphid-infested wild-type plants for two of the accessions. In contrast, female E. balteatus predators did not differentiate between aphid-infested transgenic or wild-type plants. CONCLUSION: The results indicate that the genetic engineering of plants to modify their emission of VOCs holds considerable promise for facilitating biological control of herbivores. Validation for crop plants is a necessary next step to assess the usefulness of modified volatile emission in integrated pest management.

A long-term experiment on the effect of chitin addition to soil on the suppression of soil-borne pathogens was set up and monitored for eight years in an experimental field, Vredepeel, the Netherlands. Chitinous matter obtained from shrimps was added to soil top layers at two different occasions and the suppressiveness of soil towards Verticillium dahliae as well as plant-pathogenic nematodes was assessed, in addition to analyses of the abundances and community structures of members of the soil microbiota. The data revealed that chitin amendment had raised the suppressiveness of soil, in particular towards Verticillium dahliae, 9 months after the (second) treatment, extending to two years following treatment. Moreover, major effects of the added chitin on the soil microbial communities were detected. First, shifts in both the abundances and structures of the chitin-treated soil microbial communities were found, both of total soil bacteria and fungi. In addition, the abundances and structures of soil actinobacteria and Oxalobacteriaceae were affected by chitin. At the functional gene level, the abundance of specific (family-18 glycoside hydrolase) chitinase genes carried by the soil bacteriota also revealed upshifts as a result of the added chitin. The effects of chitin noted for the Oxalobacteriaceae were specifically related to significant upshifts in the abundances of the species Duganella violaceinigra and Massilia plicata. These effects of chitin persisted over the time of the experiment.

Chitin amendment is a promising soil management strategy that may enhance the suppressiveness of soil toward plant pathogens. However, we understand very little of the effects of added chitin, including the putative successions that take place in the degradative process. We performed an experiment in moderately acid soil in which the level of chitin, next to the pH, was altered. Examination of chitinase activities revealed fast responses to the added crude chitin, with peaks of enzymatic activity occurring on day 7. PCR-denaturing gradient gel electrophoresis (DGGE)-based analyses of 16S rRNA and chiA genes showed structural changes of the phylogenetically and functionally based bacterial communities following chitin addition and pH alteration. Pyrosequencing analysis indicated (i) that the diversity of chiA gene types in soil is enormous and (i) that different chiA gene types are selected by the addition of chitin at different prevailing soil pH values. Interestingly, a major role of Gram-negative bacteria versus a minor one of Actinobacteria in the immediate response to the added chitin (based on 16S rRNA gene abundance and chiA gene types) was indicated. The results of this study enhance our understanding of the response of the soil bacterial communities to chitin and are of use for both the understanding of soil suppressiveness and the possible mining of soil for novel enzymes.

Endophytic Pseudomonas aeruginosa strain BP35 was originally isolated from black pepper grown in the rain forest in Kerala, India. Strain PaBP35 was shown to provide significant protection to black pepper against infections by Phytophthora capsici and Radopholus similis. For registration and implementation in disease management programmes, several traits of PaBP35 were investigated including its endophytic behaviour, biocontrol activity, phylogeny and toxicity to mammals. The results showed that PaBP35 efficiently colonized black pepper shoots and displayed a typical spatiotemporal pattern in its endophytic movement with concomitant suppression of Phytophthora rot. Confocal laser scanning microscopy revealed high populations of PaBP35::gfp2 inside tomato plantlets, supporting its endophytic behaviour in other plant species. Polyphasic approaches to genotype PaBP35, including BOX-PCR, recN sequence analysis, multilocus sequence typing and comparative genome hybridization analysis, revealed its uniqueness among P.¿aeruginosa strains representing clinical habitats. However, like other P.¿aeruginosa strains, PaBP35 exhibited resistance to antibiotics, grew at 25–41°C and produced rhamnolipids and phenazines. PaBP35 displayed strong type II secretion effectors-mediated cytotoxicity on mammalian A549 cells. Coupled with pathogenicity in a murine airway infection model, we conclude that this plant endophytic strain is as virulent as clinical P.¿aeruginosa strains. Safety issues related to the selection of plant endophytic bacteria for crop protection are discussed.

International audience; Semi-natural structures of agricultural landscapes are known to provide important habitat functions for natural enemies of crop herbivores. Although several functions such as resource and shelter are related to plant species, the role of the floristic composition is not well understood. Using a correlative approach, we analysed interactions between plant functional groups of spontaneous field margin vegetation and the regulation of crop herbivores. We further compared the influence of the local plant community with that of landscape structure at different scales (50-500 m). We used Brassica oleracea crop plants as phytometers planted into field margins of 48 fields and observed correlations of explanatory variables (vegetation, landscape) and response variables (herbivore and natural enemy abundance, crop damage) for two consecutive years (24 fields x year(-1)). We observed significant positive and negative correlations between plant functional groups and variables related to crop herbivore regulation. In general, plant cover based-models provided a better fit than diversity-based models. Entomophilous flowering plants providing nectar showed a positive correlation to ladybird abundance and aphid parasitism and a negative relationship with major crop plant damage caused by leaf-chewing invertebrates. Cover and diversity of wild Brassicaceae species were positively correlated with flea beetle damage and abundance. Most correlations with plant functional groups remained significant after fitting variables of landscape structure. The density of semi natural habitats showed predominantly positive correlations to crop herbivore regulation, whereas the density of crops was negatively correlated. The number of significant relationships decreased from natural enemy to herbivore abundance and to plant damage. Our results demonstrate the relative importance of field margin vegetation for crop herbivore regulation. The differential responses to plant functional groups support the idea that designing specific plant mixtures may improve biological control. Additional effects of landscape variables suggest that a combination of local and landscape-scale management is the most promising strategy to optimise this ecosystem service. (C) 2016 Elsevier B.V. All rights reserved.

Chitin is a promising soil amendment for improving soil quality, plant growth and plant resilience. The objectives of this study were twofold. First, to study the effect of chitin mixed in potting soil on lettuce growth and on the survival of two zoonotic bacterial pathogens, Escherichia coli O157:H7 and Salmonella enterica on the lettuce leaves. Second, to assess the related changes in the microbial lettuce rhizosphere, using phospholipid fatty acid (PLFA) analysis and amplicon sequencing of a bacterial 16S rRNA gene fragment and the fungal ITS2. As a result of chitin addition, lettuce fresh yield weight was significantly increased. S. enterica survival in the lettuce phyllosphere was significantly reduced. The E. coli O157:H7 survival was also lowered, but not significantly. Moreover, significant changes were observed in the bacterial and fungal community of the lettuce rhizosphere. PLFA analysis showed a significant increase in fungal and bacterial biomass. Amplicon sequencing showed no increase in fungal and bacterial biodiversity, but relative abundances of the bacterial phyla Acidobacteria, Verrucomicrobia, Actinobacteria, Bacteroidetes, and Proteobacteria and the fungal phyla Ascomycota, Basidiomycota, and Zygomycota were significantly changed. More specifically, a more than tenfold increase was observed for operational taxonomic units (OTUs) belonging to the bacterial genera Cellvibrio, Pedobacter, Dyadobacter, and Streptomyces and to the fungal genera Lecanicillium and Mortierella. These genera include several species previously reported to be involved in biocontrol, plant growth promotion, the nitrogen cycle and chitin degradation. These results enhance the understanding of the response of the rhizosphere microbiome to chitin amendment. Moreover, this is the first study to investigate the use of soil amendments to control the survival of S. enterica on plant leaves.

Multiparasitism (females of multiple species parasitizing the same host) is a ubiquitous phenomenon in parasitoids, yet the role of within-host competition has been mostly ignored in multiparasitoid-host models. Here we study the effect of varying the degree of competition at different life stages: competition over oviposition sites (between-adult competition) and larval competition over resources within the host (within-host competition). We adapt a Nicholson-Bailey model to allow for varying levels of between-adult competition (varying the overlap in species distributions) and within-host competition (varying the number of offspring that can successfully emerge from a host). Surprisingly, while stronger between-adult competition reduces coexistence, stronger within-host competition promotes it. Asymmetric between-adult competition (a fecundity difference between the two species) reduces coexistence when compared to symmetric competition; this can be counteracted by asymmetric within-host competition (within-host competitive advantage of the lower-fecundity species), but only when within-host competition is strong and the correlation between the parasitoids' distributions is intermediate. We discuss our results in the context of the interaction between two parasitoid species, Nasonia vitripennis and Nasonia giraulti, which have strongly correlated distributions and high levels of multiparasitism in the field. We conclude that either low or asymmetric within-host competition is unlikely to explain their coexistence.

International audience; Classical biological control-the introduction of exotic species to permanently control pests-offers an applied framework to test ecological and evolutionary hypotheses derived from invasion biology. One such hypothesis is that intraspecific hybridization can facilitate invasions because hybrids express higher phenotypic mean and/or variance than their parents. We tested this hypothesis using the parasitoid wasp Psyttalia lounsburyi, a candidate biocontrol agent for the olive fly Bactrocera oleae. Under laboratory conditions, we found marked differentiations between two populations of wasps, from South Africa and Kenya, in terms of life history strategies. South African females were better reproducers than Kenyan females, but the opposite was observed for males. Reaction norms showed different effects of developmental temperature on fecundity depending on the genotype. However, neither heterosis nor hybrid breakdown were observed. Hence, in this system, sex-specific effects of hybridization and genotype-by-environment interactions jeopardize any straightforward prediction on the fitness of hybrids. Therefore, our paper contributes to tone down the hybrid advantage hypothesis in invasion biology.

Possibilities for biocontrol of biovar 3 Dickeya sp. in potato were investigated, using bacteria from rotting potato tissue isolated by dilution plating on nonselective agar media. In a plate assay, 649 isolates were screened for antibiosis against Dickeya sp. IPO2222 and for the production of siderophores. Forty-one isolates (6Æ4%) produced antibiotics and 112 isolates (17Æ3%) produced siderophores. A selection of 41 antibiotic-producing isolates and 41 siderophore-producing isolates were tested in a potato slice assay for control of the Dickeya sp. Isolates able to reduce rotting of potato tuber tissue by at least 50% of the control were selected. Isolates were characterized by 16S rDNA analysis as Bacillus, Pseudomonas, Rhodococcus, Serratia, Obesumbacterium and Lysinibacillus genera. Twenty-three isolates belonging to different species and genera, 13 producing antibiotics and 10 producing siderophores, were further characterized by testing acyl-homoserine lactone (AHL) production, quorum quenching, motility, biosurfactant production, growth at low (4Æ0) and high (10Æ0) pH, growth at 10_C under aerobic and anaerobic conditions and auxin production. In replicated greenhouse experiments, four selected antagonists based on the in vitro tests were tested in planta using wounded or intact minitubers of cv. Kondor subsequently inoculated by vacuum infiltration with an antagonist and a GFP (green fluorescent protein)-tagged biovar 3 Dickeya sp. strain. A potato endophyte A30, characterized as S. plymuthica, protected potato plants by reducing blackleg development by 100% and colonization of stems by Dickeya sp. by 97%. The potential use of S. plymuthica A30 for the biocontrol of Dickeya sp. is discussed

Interactions between Serratia plymuthica A30 and a blackleg-causing biovar 3 Dickeya sp. were examined. In a potato slice assay, S. plymuthica A30 inhibited tissue maceration caused by Dickeya sp. IPO2222 when co-inoculated at a density at least 10 times greater than that of the pathogen. In glasshouse experiments, population dynamics of the antagonist and of the pathogen in planta were studied by dilution plating and confocal laser scanning microscopy (CLSM) using fluorescent protein-tagged strains. Pathogen-free minitubers were vacuum-infiltrated with DsRed-tagged Dickeya sp. IPO2222 and superficially treated during planting with a water suspension containing GFP-tagged S. plymuthica A30. A30 reduced the blackleg incidence from 55% to 0%. Both the pathogen and the antagonist colonized the seed potato tubers internally within 1 day post-inoculation (dpi). Between 1 and 7 dpi, the population of A30 in tubers increased from 101 to c. 103 CFU g)1 and subsequently remained stable until the end of the experiment (28 dpi). Populations of A30 in stems and roots increased from c. 102 to c. 104 CFU g)1 between 7 and 28 dpi. Dilution plating and CLSM studies showed that A30 decreased the density of Dickeya sp. populations in plants. Dilution plating combined with microscopy allowed the enumeration of strain A30 and its visualization in the vascular tissues of stem and roots and in the pith of roots, as well as its adherence to and colonization of the root surface. The implications of these finding for the use of S. plymuthica A30 as a biocontrol agent are discussed

Phoretic insects utilize other animals to disperse to new environments. We recently discovered how egg parasitoids use an exciting phoretic strategy to reach egg-laying sites of their butterfly hosts. In the laboratory, female Trichogramma wasps detect and mount mated female cabbage white butterflies that emit an anti-aphrodisiac pheromone. Hardly any information exists about the natural occurrence of phoresy in wasps of this genus. Therefore, we monitored the presence of phoretic Trichogramma wasps on lepidopteran hosts in the field. Only female wasps were found at low prevalence on six lepidopteran species. Wasps were mostly found on female hosts and mainly on abundant solitary host species. This is the first report of phoretic Trichogramma wasps on butterflies in nature. We suggest that phoresy is only one of several strategies used by these polyphagous egg parasitoids. The evolution of phoresy is discussed in relation to the nutritional ecology of egg parasitoids.

Abstract The use of organic soil amendments, such as green manures, animal manures, composts or slurries, certainly has many advantageous aspects for soil quality and is suggested as a promising tool for the management of plant-parasitic nematodes. However, during a recent literature survey we also found numerous studies reporting an increase of plant-parasitic nematodes after the use of organic amendments. Therefore, we critically re-evaluated the usefulness of organic amendments for nematode management and suggest possible mechanisms for a stimulation of plant-parasitic nematodes, as well as mechanisms that might be causing a reduction of plant-parasitic nematodes. In addition, we also elucidate a possible mechanism that might be responsible for the observed overall positive effects of organic amendments on crop yields. It is likely that a significant part of this is, inter alia, due to the proliferation of non-pathogenic, free-living nematodes and their overall positive effects on soil microbial populations, organic matter decomposition, nutrient availability, plant morphology and ecosystem stability.

The parasitoid wasp Apanteles taragamae is a promising candidate for the biological control of the legume pod borer Maruca vitrata, which recently has been introduced into Benin. The effects of volatiles from cowpea and peabush flowers and Maruca vitrata larvae on host selection behavior of the parasitoid Apanteles taragamae were investigated under laboratory conditions by using a Y-tube olfactometer. Naïve and oviposition-experienced female wasps were given a choice between several odor sources that included (1) uninfested, (2) Maruca vitrata-infested, and (3) mechanically damaged cowpea flowers, as well as (4) stem portions of peabush plants carrying leaves and flowers, (5) healthy M. vitrata larvae, and moribund (6), and live (7) virus-infected M. vitrata larvae. Responses of naïve and oviposition-experienced female wasps did not differ for any of the odor source combinations. Wasps were significantly attracted to floral volatiles produced by cowpea flowers that had been infested with M. vitrata larvae and from which the larvae had been removed. Apanteles taragamae females also were attracted to Maruca vitrata-infested flowers after removal of both the larvae and their feces. Female wasps discriminated between volatiles from previously infested flowers and mechanically damaged flowers. Uninfested cowpea flowers attracted only oviposition-experienced wasps that had received a rewarding experience (i.e. the parasitization of two M. vitrata larvae feeding on cowpea flowers) before the olfactometer test. Wasps also were attracted to uninfested leaves and flowers of peabush. Moreover, they were also attracted to healthy and live virus-infected M. vitrata larvae, but not when the latter were moribund. Our data show that, similarly to what has been extensively been reported for foliar volatiles, flowers of plants also emit parasitoid-attracting volatiles in response to being infested with an herbivore.

Naturally occurring insect viruses can modify the behaviour of infected insects and thereby modulate virus transmission. Modifications of the virus genome could alter these behavioural effects. We studied the distance moved and the position of virus-killed cadavers of fourth instars of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) infected with a wild-type genotype of H. armigera nucleopolyhedrovirus (HaSNPV) or with one of two recombinant genotypes of this virus on cotton plants. The behavioural effects of virus infection were examined both in larvae infected with a single virus genotype, and in larvae challenged with mixtures of the wild-type and one of the recombinant viruses. An egt-negative virus variant caused more rapid death and lower virus yield in fourth instars, but egt-deletion did not produce consistent behavioural effects over three experiments, two under controlled glasshouse conditions and one in field cages. A recombinant virus containing the AaIT-(Androctonus australis Hector) insect-selective toxin gene, which expresses a neurotoxin derived from a scorpion, caused faster death and cadavers were found lower down the plant than insects infected with unmodified virus. Larvae that died from mixed infections of the AaIT-expressing recombinant and the wild-type virus died at positions significantly lower, compared to infection with the pure wild-type viral strain. The results indicate that transmission of egt-negative variants of HaSNPV are likely to be affected by lower virus yield, but not by behavioural effects of egt gene deletion. By contrast, the AaIT recombinant will produce lower virus yields as well as modified behaviour, which together can contribute to reduced virus transmission under field conditions. In addition, larvae infected with both the wild-type virus and the toxin recombinant behaved as larvae infected with the toxin recombinant only, which might be a positive factor for the risk assessment of such toxin recombinants in the environment.

One of the key issues in compost research is to assess when the compost has reached a mature stage. The maturity status of the compost determines the quality of the final soil amendment product. The nematode community occurring in a Controlled Microbial Composting (CMC) process was analyzed with the objective of assessing whether the species composition could be used as a bio-indicator of the compost maturity status. The results obtained here describe the major shifts in species composition that occur during the composting process. Compared to terrestrial ecosystems, nematode succession in compost differs mainly in the absence of K-strategists and numerical importance of diplogastrids. At the beginning of the composting process (thermophilic phase), immediately after the heat peak, the nematode population is primarily built by bacterial feeding enrichment opportunists (cp-1) (Rhabditidae, Panagrolaimidae, Diplogastridae) followed by the bacterial-feeding general opportunists (cp-2) (Cephalobidae) and the fungal-feeding general opportunists (Aphelenchoididae). Thereafter, during the cooling and maturation stage, the bacterial-feeding-predator opportunistic nematodes (Mononchoides sp.) became dominant. Finally, at the most mature stage, the fungal-feeding Anguinidae (mainly Ditylenchus filimus) were most present. Both, the Maturity Index (MI) and the fungivorous/bacterivorous ratio (f/b ratio), increase as the compost becomes more mature (ranging, respectively, from 1 to 1.86 and from 0 to 11.90). Based on these results, both indices are suggested as potential suitable tools to assess compost maturity.

Invasive species represent one of the most important threats to biodiversity worldwide, with consequences for ecosystem functioning and the delivery of important ecological services to society. Several hypotheses have been generated to explain the success of exotic plants in their new ranges, with escape from their old natural enemies, such as pathogens and herbivores (the ‘enemy release hypothesis’) and novel defensive chemistry (the ‘novel weapons hypothesis’) receiving considerable attention. Thus far, virtually all studies of exotic plants and insects have been conducted in a strictly bi-trophic framework involving plants and herbivores. On the other hand, it has been argued that a better understanding of the forces regulating community structure and function should include natural enemies of the herbivores. Furthermore, indirect interactions between organisms in the plant roots (below-ground) and shoots (above-ground) are known to strongly effect the behaviour and performance of consumers in the opposite ‘compartment’. Here, we discuss a range of physiological, evolutionary and ecological aspects of plant–herbivore-natural enemy interactions involving exotic plants. Further, interactions between soil and above-ground organisms are explored with respect to studies with exotic plants. We argue that it is important to link population and community ecology to individual-level variation in the physiology and behaviour of insects across several trophic levels in studies with invasive plants. Future research with invaders should also aim to integrate physically separated compartments (e.g. plant roots and shoots). This will facilitate a more complete understanding of the factors underlying the success (or failure) of exotic plants to spread and become dominant in their new ranges. Moreover, these data will also help to unravel the relative importance of top-down and bottom-up processes in regulating communities in which exotic plants have become established. Lastly, we discuss consequences for conservation.

Direct observation of searching patterns and residence times of Trichogramma brassicae Bezdenko on maize plants were made at 18 degrees C and 25 degrees C. Temperature had a strong effect on the residence times: parasitoids spent an average of 44.9 minutes on the plant at 18 degrees C and 20.8 minutes at 25 degrees C. Observations on single plants showed that parasitoids mainly walked from one leaf to another. The leaf part closest to the stem was the most often visited and longest searched leaf area. At 18 degrees C, many parasitoids went to the lower leaf side and stopped searching. Leaf level, leaf side or size of the leaf where the parasitoid landed had no effect on residence time. Although about 20% of total searching time was spent in following the leaf edge and mid rib, Ostrinia (Hubner) egg masses touching the mid rib were not found more often than those away from the mid rib. Host finding was also not influenced by the position of egg masses relative to the stem, or by the leaf height. Naturally laid egg masses were found twice as often as artificially placed egg masses. The scales that O. nubilalis adults left in small patches on the plant seemed to be as effective a cue as host cues artificially confined to a small area around the host egg mass. When maize plants were covered with fine netting and a number of O. nubilalis kept close to the plant for one night in such a way that the moths were unable to touch the plants, host finding and residence times of T. brassicae were significantly higher on these plants than on untreated plants. This indicates that volatile cues left by O. nubilalis had an arresting effect and were used in host finding by T. brassicae.

Political efforts are made in the European Union (EU) to reduce pesticide use and to increase the implementation of integrated pest management (IPM). Within the EU project ENDURE, research priorities on pesticide reduction are defined. Using maize, one of the most important crops in Europe, as a case study, we identified the most serious weeds, arthropod pests, and fungal diseases as well as classes and amounts of pesticides applied. Data for 11 European maize growing regions were collected from databases, publications and expert estimates. Silage maize dominates in northern Europe and grain production in central and southern Europe. Crop rotations range from continuous growing of maize over several years to well-planned rotation systems. Weeds, arthropod pests and fungal diseases cause economic losses in most regions, even though differences exist between northern countries and central and southern Europe. Several weed and arthropod species cause increasing problems, illustrating that the goal of reducing chemical pesticide applications is challenging. Pesticides could potentially be reduced by the choice of varieties including genetically modified hybrids, cultural control including crop rotation, biological control, optimized application techniques for chemicals, and the development of more specific treatments. However, restrictions in the availability of alternative pest control measures, farm organization, and the training and knowledge of farmers need to be overcome before the adoption of environmentally friendly pest control strategies can reduce chemical pesticides in an economically competitive way. The complex of several problems that need to be tackled simultaneously and the link between different control measures demonstrates the need for IPM approaches, where pest control is seen in the context of the cropping system and on a regional scale. Multicriteria assessments and decision support systems combined with pest monitoring programs may help to develop region-specific and sustainable strategies that are harmonized within a EU framework

Ralstonia solanacearum race 3 biovar 2 (phylotype II) is the causal agent of bacterial wilt of tomato, the most destructive bacterial disease of this crop in Ethiopia for which no effective control measures are available. In this study, the effects of amending topsoil with three different levels (1, 5 and 10%) of cocopeat, farmyard manure (FYM) compost and green compost, and two levels of bacterial inoculations were tested on infection of tomato by R. solanacearum compared to non-inoculated treatments. Non-amended topsoil with and without R. solanacearum were included as control treatments. Survival of and infection by R. solanacearum and yield-associated agronomic responses were used for evaluation along with physico-chemical and biological characteristics of amended soils. Amendments resulted in changes in physico-chemical properties (such as electrical conductivity, organic matter content, total carbon, dissolved organic carbon, NH(4)(+), NO(3)(-)) and microbial activity of the amended soil and the effects were found to be higher at the higher rate of application. Effects on disease suppression and survival of the pathogen in the soil differed depending on amendment type and application rate. Higher disease severity was recorded in soil amended with 10% green compost compared to the control treatment. Complete suppression of R. solanacearum was observed in pots amended with 5 and 10% farm yard manure (FYM), 1% green compost and 10% cocopeat. Absence of disease at the highest rate of FYM was supported by a lower number of culturable R. solanacearum bacteria recovered from rhizosphere soil two months post-inoculation in soil amended with 10% FYM. Soil amended with 10% FYM gave higher root and above-ground dry weight. Moreover, FYM added to topsoil at 5 and 10% gave significantly higher above-ground fresh weight. This study indicated that amending topsoil with different types and rates of amendment can suppress bacterial wilt severity and pathogen survival in the soil. Amendments also enhanced tomato yield, the higher rates of amendments being the most effective except green compost at 10% which gave a 27% lower yield compared with the higher rate of FYM. Among the amendments tested, FYM at 5 or 10% would be an interesting option to manage R. solanacearum in the major tomato-growing regions of Ethiopia. However, the mechanisms of disease suppression at higher rates of FYM need to be investigated.

Although integrated pest management (IPM) strategies have been developed worldwide, further improvement of IPM effectiveness is required. The use of transgenic technology to create insect-resistant plants can offer a solution to the limited availability of highly insect-resistant cultivars. Commercially available insect-resistant transgenic crops show clear benefits for agriculture and there are many exciting new developments such as transgenic plants that enhance biological control. Effective evaluation tools are needed to ascertain that transgenic plants do not result in undesired non-target effects. If these conditions are met, there will be ample opportunities for transgenic plants to become key components of environmentally benign and durable pest management systems. Here we discuss the potential and challenges for incorporating transgenic plants in IPM.

Filamentous fungi in the genus Colletotrichum are destructive pathogens that cause disease and crop losses in plants worldwide. Taxonomy and nomenclature in the group is confusing, even to scientists working in the field, and inaccurate diagnosis of species is not uncommon. In this review, we provide a overview of the 66 Colletotrichum names that are in common use, and the 19 recently used names which are regarded as doubtful. This paper represents the first comprehensive overview of the genus in 17 years, and is the first summary treatment of Colletotrichum to incorporate data generated through DNA analysis and phylogenetic systematics. Species are listed alphabetically and annotated with their taxonomic entry, teleomorph, hosts and disease, brief summaries of taxonomic and phylogenetic research, and outstanding issues for the genus that are neccesary to stabilize species names. Sequence data and type culture collection resources are also summarized. The paper serves to provide a new starting point for usage of current names in Colletotrichum and indicates future work needed

The effects of genotype, plant growth and experimental factors (soil and year) on potato-associated bacterial communities were studied. Cultivars Achirana Inta, Desiree, Merkur and transgenic Desiree line DL12 (containing T4 lysozyme gene) were assessed in two field experiments. Cross-comparisons between both experiments were made using Desiree plants. Culture-dependent and -independent approaches were used to demonstrate effects on total bacterial, actinobacterial and Pseudomonas communities in bulk and rhizosphere soils and endospheres. PCR-denaturing gradient gel electrophoresis fingerprints prepared with group-specific primers were analyzed using multivariate analyses and revealed that bacterial communities in Achirana Inta plants differed most from those of Desiree and Merkur. No significant effects were found between Desiree and DL12 lines. Plant growth stage strongly affected different plant-associated communities in both experiments. To investigate the effect of plant-associated communities on plant health, 800 isolates from rhizospheres and endospheres at the flowering stage were tested for suppression of Ralstonia solanacearum biovar 2 and/or Rhizoctonia solani AG3. A group of isolates closely resembling Lysobacter sp. dominated in young plants. Its prevalence was affected by plant growth stage and experiment rather than by plant genotype. It was concluded that plant growth stage overwhelmed any effect of plant genotype on the bacterial communities associated with potato.