Epidermis-Specific Metabolic Engineering of Sesquiterpene Formation in Tomato Affects the Performance of Potato Aphid Macrosiphum euphorbiae

Tomato produces a number of terpenes in their glandular trichomes that contribute to host plant resistance against pests. While glandular trichomes of cultivated tomato Solanum lycopersicum primarily accumulate a blend of monoterpenes, those of the wild tomato species Solanum habrochaites produce various sesquiterpenes. Recently, we have identified two groups of sesquiterpenes in S. habrochaites accessions that negatively affect the performance and choice behavior of the potato aphid (Macrosiphum euphorbiae). Aphids are piercing-sucking herbivores that use their mouthpart to penetrate and probe plant tissues in order to ultimately access vascular tissue and ingest phloem sap. Because secondary metabolites produced in glandular trichomes can affect the initial steps of the aphid feeding behavior, introducing the formation of defensive terpenes into additional plant tissues via metabolic engineering has the potential to reduce tissue penetration by aphids and in consequence virus transmission. Here, we have developed two multicistronic expression constructs based on the two sesquiterpene traits with activity toward M. euphorbiae previously identified in S. habrochaites. Both constructs are composed of sequences encoding a prenyl transferase and a respective S. habrochaites terpene synthase, as well as enhanced green fluorescent protein as a visible marker. All three coding sequences were linked by short nucleotide sequences encoding the foot-and-mouth disease virus 2A self-processing oligopeptide which allows their co-expression under the control of one promoter. Transient expression of both constructs under the epidermis-specific Arabidopsis CER5-promoter in tomato leaves demonstrated that formation of the two sets of defensive sesquiterpenes, β-caryophyllene/α-humulene and (−)-endo-α-bergamotene/(+)-α-santalene/(+)-endo-β-bergamotene, can be introduced into new tissues in tomato. The epidermis-specific transgene expression and terpene formation were verified by fluorescence microscopy and tissue fractionation with subsequent analysis of terpene profiles, respectively. In addition, the longevity and fecundity of M. euphorbiae feeding on these engineered tomato leaves were significantly reduced, demonstrating the efficacy of this novel aphid control strategy.

Selection of Endophytic Beauveria bassiana as a Dual Biocontrol Agent of Tomato Pathogens and Pests

Endophytic fungi (EF) can enhance both plant growth and defense barriers against pests and pathogens, contributing to the reduction of chemical pesticides and fertilizers use in agriculture. Beauveria bassiana is an entomopathogenic fungus showing endophytism in several crops, often associated with a good capacity to limit the development of pests and disease agents. However, the diversity of the protective efficacy and plant response to different strains can be remarkable and needs to be carefully assessed for the successful and predictable use of these beneficial microorganisms. This study aims to select B. bassiana strains able to colonize tomato plants as endophytes as well as to control two important disease agents, Botrytis cinerea and Alternaria alternata, and the pest aphid, Macrosiphum euphorbiae. Nine wild-type isolates and one commercial strain were screened for endophytism, then further characterized for plant-growth promotion plus inhibition of disease development and pest infestation. Four isolates proved to have a good control activity against the biotic stressors tested, but only Bb716 was also able to promote plant growth. This work provides a simple workflow for the selection of beneficial EF, paving the way towards more effective use of B. bassiana in Integrate Pest Management (IPM) of tomato.

Biopesticide Evaluation from Lab to Greenhouse Scale of Essential Oils Used against Macrosiphum euphorbiae

Aphids are recognized as a major threat to economically important crops. Their control is predominantly based on synthetic insecticides that are detrimental to human health and the environment. Botanical pesticides based on essential oils (EOs) are a promising alternative. In this study, the entomotoxicity of green anise and fennel EO fumigation was tested on the potato aphid Macrosiphum euphorbiae. Three different settings of increasing scale were considered (leaflet, whole plant and greenhouse) to appraise the consistency of EO impact from controlled laboratory to greenhouse production conditions. LC50 values for green anise and fennel were 6.6 μl L−1air and 12.2 μl L−1air, respectively, based on dose-response curves in leaflet experiments but fennel EO induced phytotoxicity. EO efficiency was confirmed at the whole-plant scale. In the greenhouse experiment, fennel EO exhibited greater efficiency than at the laboratory scale equaling green anise EO efficiency but both EOs showed delayed phytotoxicity, illustrating the importance of long-term monitoring. The present study revealed the ability of both EOs to control M. euphorbiae populations under greenhouse conditions and hinted at the importance of assessing EO efficiency in realistic agronomic conditions (e.g., under the fluctuating environmental conditions usually occurring in greenhouses).

Field Evaluation of Cypermethrin, Imidacloprid, Teflubenzuron and Emamectin Benzoate against Pests of Quinoa (Chenopodium quinoa Willd.) and Their Side Effects on Non-Target Species

During the last few years, quinoa, a traditional Andean crop, has been cultivated at low elevations where pest pressure is high and farmers resort to intensive use of insecticides. This field study investigated the impact of four insecticides (cypermethrin, imidacloprid, teflubenzuron and emamectin benzoate) on insect pests of quinoa and their side effects on the arthropod community at the coastal level of Peru, by analysing the species composition, species diversity and population density. The arthropod community was examined with pitfall traps (for ground dwelling species), plant samplings (for pests and their natural enemies that inhabit the crop), and yellow pan traps (to catch flying insects). The results demonstrated that Macrosiphum euphorbiae, Frankliniella occidentalis and Spoladea recurvalis were efficiently controlled by cypermethrin and imidacloprid; the latter compound also showed long-term effects on Nysius simulans. Teflubenzuron and emamectin benzoate proved to be efficient to control S. recurvalis. Imidacloprid had the strongest adverse effects on the arthropod community in terms of species diversity, species composition and natural enemy density as compared to the other insecticides. Findings of this study may assist farmers intending to grow quinoa at the coastal level in selecting the most appropriate insecticides under an integrated pest management approach.

Post-Dropping Behavior of Potato Aphids (Macrosiphum euphorbiae)

Dropping behavior is an effective antipredator defense utilized by many insects including aphids, which drop from plants to lower plant parts or underlying substrates to avoid attack from predatory invertebrates. While research commonly focusses on triggers of dropping, less attention is given to what happens to prey individuals following escape drops. In this study, the duration of tonic immobility, recovery rates, and cases of “instant recovery” (re-clinging to lower plant parts) exhibited by potato aphids (Macrosiphum euphorbiae) that dropped from potted seedlings in response to introduced ladybird (Adalia bipunctata) adults, lacewing (Chrysoperla carnea) larvae, and a standardized tactile stimulus were investigated in relation to a range of environmental factors. Air temperature had a negative correlation with the duration of post-dropping tonic immobility; as temperature increased, time spent motionless decreased. Aphids also showed a pattern of increased recovery rate at higher temperatures. Aphids may be selected to move off the substrate quicker to avoid risks of overheating/desiccation at higher temperatures; and/or higher body temperature facilitates locomotion. Stimulus type also influenced recovery rate back to the original seedling, with aphids generally recovering after the standardized stimulus quicker than after dropping triggered by a real predator. Considering cases of instant recovery onto lower-reaches of the host seedling, seedling height influenced the likelihood of re-clinging, with aphids that managed to instantly recover dropping from, on average, taller seedlings than aphids that dropped to the substrate. Plant architecture could mitigate the costs of dropping for aphids, but further studies quantifying understory foliage cover are needed.

Temperature Differentially Influences the Capacity of Trichoderma Species to Induce Plant Defense Responses in Tomato Against Insect Pests

Species of the ecological opportunistic, avirulent fungus, Trichoderma are widely used in agriculture for their ability to protect crops from the attack of pathogenic fungi and for plant growth promotion activity. Recently, it has been shown that they may also have complementary properties that enhance plant defense barriers against insects. However, the use of these fungi is somewhat undermined by their variable level of biocontrol activity, which is influenced by environmental conditions. Understanding the source of this variability is essential for its profitable and wide use in plant protection. Here, we focus on the impact of temperature on Trichoderma afroharzianum T22, Trichoderma atroviride P1, and the defense response induced in tomato by insects. The in vitro development of these two strains was differentially influenced by temperature, and the observed pattern was consistent with temperature-dependent levels of resistance induced by them in tomato plants against the aphid, Macrosiphum euphorbiae, and the noctuid moth, Spodoptera littoralis. Tomato plants treated with T. afroharzianum T22 exhibited enhanced resistance toward both insect pests at 25°C, while T. atroviride P1 proved to be more effective at 20°C. The comparison of plant transcriptomic profiles generated by the two Trichoderma species allowed the identification of specific defense genes involved in the observed response, and a selected group was used to assess, by real-time quantitative reverse transcription PCR (qRT-PCR), the differential gene expression in Trichoderma-treated tomato plants subjected to the two temperature regimens that significantly affected fungal biological performance. These results will help pave the way toward a rational selection of the most suitable Trichoderma isolates for field applications, in order to best face the challenges imposed by local environmental conditions and by extreme climatic shifts due to global warming.

Assessment of potential invasion for six phytophagous quarantine pests in Taiwan

Pest risk assessment is typically performed by expert taxonomists using a pest’s biological data. However, the biological data or expert taxonomists may be difficult to find. Here, we used species distribution modelling to predict potential invasion in which phytophagous quarantine pests survive in Taiwan; the pests (unrecorded yet in Taiwan) included were three notorious quarantine whiteflies (Crenidorsum aroidephagus, Aleurothrixus trachoides, and Paraleyrodes minei) and three aphids (Nasonovia ribisnigri, Macrosiphum euphorbiae, and Viteus vitifoliae). In brief, maximum entropy modelling (MaxEnt) was used to predict the suitability of the pests’ habitats under certain climatic conditions, and then receiver operating characteristic curve analysis was performed (to verify the prediction result). We then analysed environmental variables affecting the habitat suitability and matched them with Taiwan’s crop cultivation areas for the assessment of potential invasion. We observed that the habitat suitability of the cultivation areas of host plants was low for C. aroidephagus, A. trachoides, and N. ribisnigri but was high for the remaining three species. Moreover, precipitation of coldest quarter negatively affected habitat suitability for C. aroidephagus, P. minei, N. ribisnigri, and M. euphorbiae. Seasonal temperature changes also negatively affected the habitat suitability for A. trachoides. This is the first study to demonstrate the use of species distribution modelling as the preliminary step for the pest risk assessment of these emerging pests with limited biological data before their invasion.

Soil microbial diversity impacts plant microbiota more than herbivory

Interactions between plants and microbiomes play a key role in ecosystem functioning and are of broad interest due to their influence on nutrient cycling and plant protection. However, we do not yet have a complete understanding of how plant microbiomes are assembled. Here, we tested and quantified the effect of different factors driving the diversity and composition of plant-associated microbial communities. We manipulated soil microbial diversity (high or low diversity), plant species (Solanum tuberosum or Solanum vernei), and herbivory (presence or absence of a phloem-feeding insect Macrosiphum euphorbiae), and found that soil microbial diversity influenced the herbivore-associated microbiome composition, but also plant species and herbivory influenced the soil microbiome composition. We quantified the relative strength of these effects and demonstrated that the initial soil microbiome diversity explained the most variation in plant- and herbivore-associated microbial communities. Our findings strongly suggest that soil microbial community diversity is a driver of the composition of multiple associated microbiomes (plant and insect), and this has implications for the importance of management of soil microbiomes in multiple systems.