Nicolai V. Meyling

Detection and avoidance of an entomopathogenic fungus by a generalist insect predator

Abstract.  1. Increasing evidence suggests that insects can assess their environment based on cues related to mortality risks to themselves or their offspring. Limited knowledge is available on such abilities in relation to entomopathogenic fungi, which can cause significant mortality in insect populations. In laboratory bioassays, the ability of the generalist predator Anthocoris nemorum L. (Heteroptera: Anthocoridae) to detect the presence of its natural enemy, the fungal pathogen Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) was investigated.

Community composition, host range and genetic structure of the fungal entomopathogen Beauveria in adjoining agricultural and seminatural habitats

Although intensively investigated for biological control of insect pests, little is known about the ecology of the fungal entomopathogenic genus Beauveria in natural or agricultural habitats. In this study, we used molecular phylogenetic and genotypic information to infer species diversity, reproductive potential and genetic structure of Beauveria occurring within a single arable field and bordering hedgerow in Denmark. Isolates were sampled from cultivated field and hedgerow soils, from insects harbouring latent fungal infections, and from the phylloplanes of three plant species common in the hedgerow flora. A nuclear phylogeny of this local Beauveria assemblage resolved seven phylogenetic species, including (i) five phylogenetic species within Beauveria bassiana sensu stricto; (ii) Clade C, a taxonomically uncharacterized species that is morphologically indistinguishable but phylogenetically distant from B. bassiana s.s.; and (iii) Beauveria brongniartii. All seven species were present throughout the hedgerow habitat, including as infections in insects. Significantly, only B. bassiana s.s. phylogenetic species Eu_1 was isolated from tilled soils. Mating type polymerase chain reaction assays demonstrated that all five B. bassiana s.s. phylogenetic species possess bipolar outcrossing mating systems. Of these, only the Eu_1 population contained two mating types; however, a 31:2 skew in MAT1:MAT2 mating types suggests a low frequency of sexual reproduction in this population. The four remaining B. bassiana s.s. phylogenetic species were fixed for single mating types and these populations are evidently clonal. Multilocus microsatellite genotyping revealed polymorphism in all five phylogenetic species of B. bassiana s.s.; however, all show evidence of clonal genetic structure.

Evolutionary Interaction Networks of Insect Pathogenic Fungi

Lineages of insect pathogenic fungi are concentrated in three major clades: Hypocreales (several genera), Entomophthoromycota (orders Entomophthorales and Neozygitales), and Onygenales (genus Ascosphaera). Our review focuses on aspects of the evolutionary biology of these fungi that have remained underemphasized in previous reviews. To ensure integration with the better-known domains of insect pathology research, we followed a conceptual framework formulated by Tinbergen, asking complementary questions on mechanism, ontogeny, phylogeny, and adaptation. We aim to provide an introduction to the merits of evolutionary approaches for readers with a background in invertebrate pathology research and to make the insect pathogenic fungi more accessible as model systems for evolutionary biologists. We identify a number of questions in which fundamental research can offer novel insights into the evolutionary forces that have shaped host specialization and life-history traits such as spore number and size, somatic growth rate, toxin production, and interactions with host immune systems.

Principles from community and metapopulation ecology: application to fungal entomopathogens

Fungal entomopathogens are often studied within the context of their use for biological control, yet these natural enemies are also excellent subjects for studies of ecological interactions. Here, we present selected principles from community ecology and discuss these in relation to fungal entomopathogens. We discuss the relevance of apparent competition, food web construction, intraguild predation and density-mediated and trait-mediated indirect effects. Although current knowledge of community interactions involving fungal entomopathogens are limited, fungal entomopathogens can be important, interactive members of communities and the activities of fungal entomopathogens should be evaluated in the context of ecological principles. We also discuss aspects of metapopulation ecology and the application of these principles to fungal entomopathogens. Knowledge of ecological interactions is crucial if we are to understand and predict the effects of fungal entomopathogens on host populations and understand the interactions among fungal entomopathogens and other organisms in the communities in which they occur.

Factors Affecting Vegetable Growers’ Exposure to Fungal Bioaerosols and Airborne Dust

We have quantified vegetable growers’ exposure to fungal bioaerosol components including (1→3)-β-d-glucan (β-glucan), total fungal spores, and culturable fungal units. Furthermore, we have evaluated factors that might affect vegetable growers’ exposure to fungal bioaerosols and airborne dust. Investigated environments included greenhouses producing cucumbers and tomatoes, open fields producing cabbage, broccoli, and celery, and packing facilities. Measurements were performed at different times during the growth season and during execution of different work tasks. Bioaerosols were collected with personal and stationary filter samplers. Selected fungal species (Beauveria spp., Trichoderma spp., Penicillium olsonii, and Penicillium brevicompactum) were identified using different polymerase chain reaction-based methods and sequencing. We found that the factors (i) work task, (ii) crop, including growth stage of handled plant material, and (iii) open field versus greenhouse significantly affected the workers’ exposure to bioaerosols. Packing of vegetables and working in open fields caused significantly lower exposure to bioaerosols, e.g. mesophilic fungi and dust, than harvesting in greenhouses and clearing of senescent greenhouse plants. Also removing strings in cucumber greenhouses caused a lower exposure to bioaerosols than harvest of cucumbers while removal of old plants caused the highest exposure. In general, the exposure was higher in greenhouses than in open fields. The exposures to β-glucan during harvest and clearing of senescent greenhouse plants were very high (median values ranging between 50 and 1500 ng m−3) compared to exposures reported from other occupational environments. In conclusion, vegetable growers’ exposure to bioaerosols was related to the environment, in which they worked, the investigated work tasks, and the vegetable crop.

Two Anthocoris bugs as predators of glasshouse aphids – voracity and prey preference

Voracity and prey preference were evaluated for adult females of the predatory bugs Anthocoris nemorum (L.) and Anthocoris nemoralis (Fabricius) (Heteroptera: Anthocoridae) preying upon five species of aphids (Homoptera: Aphididae), of which Myzus persicae Sulzer, Aulacorthum solani (Kaltenbach), Macrosiphum euphorbiae (Thomas), and Aphis gossypii Glover are common pests in Danish glasshouse crops. Aphis fabae Scopoli was included to determine the influence of food quality on the preference of the predators, since A. fabae has proved to be of poor nutritional value to Anthocoris spp. The experiments were carried out over 24 h in climate cabinets at 20 °C, 60–70% r.h., L18:D6. The aphids were offered in equal amounts in combinations of two species in instars of comparable size. Myzus persicae served as a reference species in all combinations. Both predators accepted all five species of aphids as prey. The numbers of aphids killed per 24 h period varied between 3.7 and 18.0 for A. nemorum and between 3.6 and 12.7 for A. nemoralis. Field collected A. nemorum females, presumably in a state of reproductive diapause, killed in three of four prey combinations significantly more aphids than did ovipositing A. nemoralis females which originated from a commercial rearing. When A. nemorum females had terminated their reproductive diapause and commenced oviposition, voracity increased approximately threefold. When prey preferences were evaluated as a total number of killed prey, no difference in preference was found between the two Anthocoris species. Both predatory bugs preferred M. persicae to the other species, the most accepted alternative prey were A. gossypii, A. fabae, A. solani, and M. euphorbiae, in descending order. However, evaluating preference by number of aphids consumed, A. nemoralis showed a more pronounced preference for M. persicae, especially when combined with A. fabae. In nearly every case, A. nemoralis rejected A. fabae as a food item after killing the aphid. Thus, A. nemoralis exhibited a more specific food choice than A. nemorum. By killing and consuming different aphid species found in glasshouse crops – particularly M. persicae– both A. nemorum and A. nemoralis showed preliminary qualities as agents for the biological control of aphids.

Mortality risk from entomopathogenic fungi affects oviposition behavior in the parasitoid wasp Trybliographa rapae

Biological control of pests in agroecosystems could be enhanced by combining multiple natural enemies. However, this approach might also compromise the control efficacy through intraguild predation (IGP) among the natural enemies. Parasitoids may be able to avoid the risk of unidirectional IGP posed by entomopathogenic fungi through selective oviposition behavior during host foraging. Trybliographa rapae is a larval parasitoid of the cabbage root fly, Delia radicum. Here we evaluated the susceptibility of D. radicum and T. rapae to two species of generalist entomopathogenic fungi, Metarhizium brunneum isolate KVL 04-57 and Beauveria bassiana isolate KVL 03-90. Furthermore, T. rapae oviposition behavior was assessed in the presence of these entomopathogenic fungi either as infected hosts or as infective propagules in the environment. Both fungi were pathogenic to D. radicum larvae and T. rapae adults, but with variable virulence. When host patches were inoculated with M. brunneum conidia in a no-choice situation, more eggs were laid by T. rapae in hosts of those patches compared to control and B. bassiana treated patches. Females that later succumbed to mycosis from either fungus laid significantly more eggs than non-mycosed females, indicating that resources were allocated to increased oviposition due to perceived decreased life expectancy. When presented with a choice between healthy and fungal infected hosts, T. rapae females laid more eggs in healthy larvae than in M. brunneum infected larvae. This was less pronounced for B. bassiana. Based on our results we propose that T. rapae can perceive and react towards IGP risk posed by M. brunneum but not B. bassiana to the foraging female herself and her offspring. Thus, M. brunneum has the potential to be used for biological control against D. radicum with a limited risk to T. rapae populations.

Beauveria bassiana as an endophyte: a critical review on associated methodology and biocontrol potential

In the last decade there has been increased focus on the potential of endophytic Beauveria bassiana for the biocontrol of insect herbivores. Generally, detection of endophytes is acknowledged to be problematic and recovery method-dependent. Herein, we critically analyse the methodology reported for the detection of B. bassiana as endophytes following experimental inoculation. In light of the methodology, we further review the effects of endophytic B. bassiana on insect herbivores. Our review indicated the need for stringent protocols for surface sterilisation including thorough experimental controls. For molecular detection protocols by PCR, residual DNA from surface inocula must also be considered. The biocontrol potential of B. bassiana endophytes appears promising although both negative and neutral effects on insect herbivores were reported and there remains ambiguity with respect to the location and mode of action of the fungus in planta. We recommend that future studies adopt multiple techniques, including culture dependent and independent techniques for endophyte detection and elucidate the mechanisms involved against insect herbivores.

Dual effects of Metarhizium spp. and Clonostachys rosea against an insect and a seed-borne pathogen in wheat

BACKGROUND Crops are often prone to both insect herbivory and disease, which necessitate multiple control measures. Ideally, an efficacious biological control agent must adequately control the target organism and not be inhibited by other biological control agents when applied simultaneously. Wheat seeds infected with the plant pathogen Fusarium culmorum were treated with Metarhizium brunneum or M. flavoviride and Clonostachys rosea individually and in combination, with the expectation to control both root-feeding insects and the pathogen. Emerging roots were evaluated for disease and then placed with Tenebrio molitor larvae, which were monitored for infection. RESULTS Plant disease symptoms were nearly absent for seeds treated with C. rosea, both individually and in combination with Metarhizium spp. Furthermore, roots grown from seeds treated with Metarhizium spp. caused significant levels of fungal infection in larvae when used individually or combined with C. rosea. However, cotreated seeds showed reduced virulence towards T. molitor when compared with treatments using Metarhizium spp. only. CONCLUSIONS This study clearly shows that seed treatments with both the entomopathogenic fungus M. brunneum and the mycoparasitic fungus C. rosea can protect plant roots from insects and disease. The dual-treatment approach to biological control presented here is consistent with the ideals of IPM strategies.

Intraguild predation by Anthocoris nemorum (Heteroptera: Anthocoridae) on the aphid parasitoid Aphidius colemani (Hhymenoptera: Braconidae)

The magnitude of intraguild predation by adult females of the predator Anthocoris nemorum on immature larvae of the aphid parasitoid Aphidius colemani inside mummies of peach-potato aphids Myzus persicae was investigated under laboratory conditions in a preference experiment. Each predator consumed a mean (95% confidence limits) of 2.8 (2.1; 3.8) immature parasitoids within mummies and 3.6 (2.7; 4.6) unparasitised aphid nymphs. Thereby A. nemorum engaged in intraguild predation with A. colemani and did not exhibit prey preference between mummies and unparasitised aphids.