Specialist root herbivore modulates plant transcriptome and downregulates defensive secondary metabolites in a brassicaceous plant.

Plants face attackers aboveground and belowground. Insect root herbivores can lead to severe crop losses, yet the underlying transcriptomic responses have rarely been studied. We studied the dynamics of the transcriptomic response of Brussels sprouts (Brassica oleracea var. gemmifera) primary roots to feeding damage by cabbage root fly larvae (Delia radicum), alone or in combination with aboveground herbivory by cabbage aphids (Brevicoryne brassicae) or diamondback moth caterpillars (Plutella xylostella). This was supplemented with analyses of phytohormones and the main classes of secondary metabolites; aromatic, indole and aliphatic glucosinolates. Root herbivory leads to major transcriptomic rearrangement that is modulated by aboveground feeding caterpillars, but not aphids, through priming soon after root feeding starts. The root herbivore downregulates aliphatic glucosinolates. Knocking out aliphatic glucosinolate biosynthesis with CRISPR-Cas9 results in enhanced performance of the specialist root herbivore, indicating that the herbivore downregulates an effective defence. This study advances our understanding of how plants cope with root herbivory and highlights several novel aspects of insect-plant interactions for future research. Further, our findings may help breeders develop a sustainable solution to a devastating root pest.

Leaf-chewing herbivores affect preference and performance of a specialist root herbivore.

Plants interact with a diversity of phytophagous insects above- and belowground. By inducing plant defence, one insect herbivore species can antagonize or facilitate other herbivore species feeding on the same plant, even when they are separated in space and time. Through systemic plant-mediated interactions, leaf-chewing herbivores may affect the preference and performance of root-feeding herbivores. We studied how six different leaf-chewing herbivore species of Brassica oleracea plants affected oviposition preference and larval performance of the root-feeding specialist Delia radicum. We expected that female D. radicum flies would oviposit where larval performance was highest, in accordance with the preference-performance hypothesis. We also assessed how the different leaf-chewing herbivore species affected defence-related gene expression in leaves and primary roots of B. oleracea, both before and after infestation with the root herbivore. Our results show that leaf-chewing herbivores can negatively affect the performance of root-feeding D. radicum larvae, although the effects were relatively weak. Surprisingly, we found that adult D. radicum females show a strong preference to oviposit on plants infested with a leaf-chewing herbivore. Defence-related genes in primary roots of B. oleracea plants were affected by the leaf-chewing herbivores, but these changes were largely overridden upon local induction by D. radicum. Infestation by leaf herbivores makes plants more attractive for oviposition by D. radicum females, while decreasing larval performance. Therefore, our findings challenge the preference-performance hypothesis in situations where other herbivore species are present.

Shoot and root insect herbivory change the plant rhizosphere microbiome and affects cabbage-insect interactions through plant-soil feedback.

Plant-soil feedback (PSF) may influence plant-insect interactions. Although plant defense differs between shoot and root tissues, few studies have examined root-feeding insect herbivores in a PSF context. We examined here how plant growth and resistance against root-feeding Delia radicum larvae was influenced by PSF. We conditioned soil with cabbage plants that were infested with herbivores that affect D. radicum through plant-mediated effects: leaf-feeding Plutella xylostella caterpillars and Brevicoryne brassicae aphids, root-feeding D. radicum larvae, and/or added rhizobacterium Pseudomonas simiae WCS417r. We analyzed the rhizosphere microbial community, and in a second set of conspecific plants exposed to conditioned soil, we assessed growth, expression of defense-related genes, and D. radicum performance. The rhizosphere microbiome differed mainly between shoot and root herbivory treatments. Addition of Pseudomonas simiae did not influence rhizosphere microbiome composition. Plant shoot biomass, gene expression, and plant resistance against D. radicum larvae was affected by PSF in a treatment-specific manner. Soil conditioning overall reduced plant shoot biomass, Pseudomonas simiae-amended soil causing the largest growth reduction. In conclusion, shoot and root insect herbivores alter the rhizosphere microbiome differently, with consequences for growth and resistance of plants subsequently exposed to conditioned soil.

Challenging the Preference-Performance Hypothesis in an above-belowground insect.

The relationship between female oviposition preference and offspring performance has been a question of special interest in the study of host plant selection by phytophagous insects. The Preference-Performance Hypothesis (PPH) is one of the main hypotheses proposed to explain this relationship, stating that females should preferentially lay eggs on plants providing the best larval development. The PPH has been extensively tested on aboveground insects but its application to species with belowground larvae is still mostly unknown. In this study, the PPH was quantitatively tested in an above-belowground insect, the cabbage root fly Delia radicum. Female oviposition preference and larval performance were estimated on three brassicaceous species (Brassica oleracea, Brassica rapa, and Sinapis alba) as well as between four cultivars of B. rapa and four cultivars of S. alba. Larval performance was estimated through their survival and through three life-history traits (LHT) of emerging adults. The PPH was supported at the intraspecific scale but only in B. rapa and for some, but not all, of the life-history traits. No support for the PPH was found in S. alba as well as at the interspecific scale. This study pleads for the integration of insects with both above- and belowground life stages in the preference-performance debate. Moreover, it raises the importance of measuring several variables to estimate larval performance and to test the PPH quantitatively, both at the plant intraspecific and interspecific scales, before drawing general conclusions.

Foliar herbivory by caterpillars and aphids differentially affects phytohormonal signalling in roots and plant defence to a root herbivore.

Plant-mediated interactions are an important force in insect ecology. Through such interactions, herbivores feeding on leaves can affect root feeders. However, the mechanisms regulating the effects of above-ground herbivory on below-ground herbivores are poorly understood. Here, we investigated the performance of cabbage root fly larvae (Delia radicum) on cabbage plants (Brassica oleracea) previously exposed to above ground herbivores belonging to two feeding guilds: leaf chewing diamondback moth caterpillars (Plutella xylostella) or phloem-feeding cabbage aphids (Brevicoryne brassicae). Our study focusses on root-herbivore performance and defence signalling in primary roots by quantifying phytohormones and gene expression. We show that leaf herbivory by caterpillars, but not by aphids, strongly attenuates root herbivore performance. Above-ground herbivory causes changes in primary roots in terms of gene transcripts and metabolites involved in plant defence. Feeding by below-ground herbivores strongly induces the jasmonate pathway in primary roots. Caterpillars feeding on leaves cause a slight induction of the primary root jasmonate pathway and interact with plant defence signalling in response to root herbivores. In conclusion, feeding by a leaf chewer and a phloem feeder differentially affects root-herbivore performance, root-herbivore-induced phytohormonal signalling, and secondary metabolites.

Peptidergic control of the crop of the cabbage root fly, Delia radicum (L.) Diptera: Anthomyiidae): A role for myosuppressin.

There is much interest in targeting neuropeptide signaling for the development of new and environmentally friendly insect control chemicals. In this study we have focused attention on the peptidergic control of the adult crop of Delia radicum (cabbage root fly), an important pest of brassicas in European agriculture. The dipteran crop is a muscular organ formed from the foregut of the digestive tract and plays a vital role in the processing of food in adult flies. We have shown using direct tissue profiling by MALDI-TOF mass spectrometry that the decapeptide myosuppressin (TDVDHVFLRFamide) is present in the crop nerve bundle and that application of this peptide to the crop potently inhibits the spontaneous contractions of the muscular lobes with an IC50 of 4.4 × 10-8 M. The delivery of myosuppressin either by oral administration or by injection had no significant detrimental effect on the adult fly. This failure to elicit a response is possibly due to the susceptibility of the peptide to degradative peptidases that cleave the parent peptide to inactive fragments. Indeed, we show that the crop of D. radicum is a source of neuropeptide-degrading endo- and amino-peptidases. In contrast, feeding benzethonium chloride, a non-peptide agonist of myosuppressin, reduced feeding rate and increased the rate of mortality of adult D. radicum. Current results are indicative of a key role for myosuppressin in the regulation of crop physiology and the results achieved during this project provide the basis for subsequent studies aimed at developing insecticidal molecules targeting the peptidergic control of feeding and food digestion in this pest species.

Influence of the symbiont Wolbachia on life history traits of the cabbage root fly (Delia radicum).

Wolbachia is an endocellular bacteria infecting arthropods and nematodes and is only transmitted vertically by females via the cytoplasm of the egg. It is often a manipulator of host reproduction, causing cytoplasmic incompatibility, thelytokous parthenogenesis, feminization or male killing, which all increase the proportion of infected females in the population. However, Wolbachia can modify life history traits of the host without causing the above phenotypes and each species illustrates the variability of relationships between this remarkably versatile symbiont and its many hosts. We have measured maternal transmission and the impact of a natural Wolbachia infection in the cabbage root fly Delia radicum, a major agricultural pest. We used a population that is polymorphic for the infection to ensure similar genetic and microbiome backgrounds between groups. Maternal transmission of the infection was 100% in our sample. We found no evidence of cytoplasmic incompatibility, thelytokous parthenogenesis, feminization nor male killing. Wolbachia infection significantly reduced hatch rate in infected eggs (by 10%) but improved larvo-nymphal viability sufficiently so that infected eggs nevertheless yielded as many adults as uninfected ones, albeit with a 1.5% longer total development time. Starved and infected ovipositing females suffered significantly reduced viability (20% higher mortality during a 3-day oviposition period) than uninfected females, but mortality was not higher in starved virgin females nor in starved males, suggesting that the energetic cost of the infection is only revealed in extreme conditions. Wolbachia had no effect on egg hatch time or offspring size. The apparently 100% vertical transmission and the significant but mutually compensating effects found suggest that infection might be nearly benign in this host and might only drift slowly, which would explain why the infection rate has been stable in our laboratory (approximately 50% individuals infected) for at least 30 generations.

Development and Validation of a SPME-GC-MS Method for In situ Passive Sampling of Root Volatiles from Glasshouse-Grown Broccoli Plants Undergoing Below-Ground Herbivory by Larvae of Cabbage Root Fly, Delia radicum L.

INTRODUCTION: Research on plant root chemical ecology has benefited greatly from recent developments in analytical chemistry. Numerous reports document techniques for sampling root volatiles, although only a limited number describe in situ collection. OBJECTIVES: To demonstrate a new method for non-invasive in situ passive sampling using solid phase micro extraction (SPME), from the immediate vicinity of growing roots. METHODS: SPME fibres inserted into polyfluorotetrafluoroethylene (PTFE) sampling tubes located in situ which were either perforated, covered with stainless steel mesh or with microporous PTFE tubing, were used for non-invasive sub-surface sampling of root volatiles from glasshouse-grown broccoli. Sampling methods were compared with above surface headspace collection using Tenax TA. The roots were either mechanically damaged or infested with Delia radicum larvae. Principal component analysis (PCA) was used to investigate the effect of damage on the composition of volatiles released by broccoli roots. RESULTS: Analyses by gas chromatography-mass spectrometry (GC-MS) with SPME and automated thermal desorption (ATD) confirmed that sulphur compounds, showing characteristic temporal emission patterns, were the principal volatiles released by roots following insect larval damage. Use of SPME with in situ perforated PTFE sampling tubes was the most robust method for out-of-lab sampling. CONCLUSION: This study describes a new method for non-invasive passive sampling of volatiles in situ from intact and insect damaged roots using SPME. The method is highly suitable for remote sampling and has potential for wide application in chemical ecology/root/soil research. Copyright © 2016 John Wiley & Sons, Ltd.

Field-based Evaluation of a Novel SPME-GC-MS Method for Investigation of Below-ground Interaction between Brassica Roots and Larvae of Cabbage Root Fly, Delia radicum L.

INTRODUCTION: Collection of volatiles from plant roots poses technical challenges due to difficulties accessing the soil environment without damaging the roots. OBJECTIVES: To validate a new non-invasive method for passive sampling of root volatiles in situ, from plants grown under field conditions, using solid phase micro-extraction (SPME). METHODS: SPME fibres were inserted into perforated polytetrafluoroethene (PTFE) tubes positioned in the soil next to broccoli plants for collection of root volatiles pre- and post-infestation with Delia radicum larvae. After sample analysis by gas chromatography-mass spectrometry (GC-MS), principal component analysis (PCA) was applied to determine differences in the profiles of volatiles between samples. RESULTS: GC-MS analysis revealed that this method can detect temporal changes in root volatiles emitted before and after Delia radicum damage. PCA showed that samples collected pre- and post-infestation were compositionally different due to the presence of root volatiles induced by D. radicum feeding. Sulphur containing compounds, in particular, accounted for the differences observed. Root volatiles emission patterns post-infestation are thought to follow the feeding and developmental progress of larvae. CONCLUSION: This study shows that volatiles released by broccoli roots can be collected in situ using SPME fibres within perforated PTFE tubes under field conditions. Plants damaged by Delia radicum larvae could be distinguished from plants sampled pre-infestation and soil controls on the basis of larval feeding-induced sulphur-containing volatiles. These results show that this new method is a powerful tool for non-invasive sampling of root volatiles below-ground. Copyright © 2016 John Wiley & Sons, Ltd.

Interactions Between a Belowground Herbivore and Primary and Secondary Root Metabolites in Wild Cabbage.

Plants are attacked by both above- and belowground herbivores. Toxic secondary compounds are part of the chemical defense arsenal of plants against a range of antagonists, and are subject to genetic variation. Plants also produce primary metabolites (amino acids, nutrients, sugars) that function as essential compounds for growth and survival. Wild cabbage populations growing on the Dorset coast of the UK exhibit genetically different chemical defense profiles, even though they are located within a few kilometers of each other. As in other Brassicaceae, the defensive chemicals in wild cabbages constitute, among others, secondary metabolites called glucosinolates. Here, we used five Dorset populations of wild cabbage to study the effect of belowground herbivory by the cabbage root fly on primary and secondary chemistry, and whether differences in chemistry affected the performance of the belowground herbivore. There were significant differences in total root concentrations and chemical profiles of glucosinolates, amino acids, and sugars among the five wild cabbage populations. Glucosinolate concentrations not only differed among the populations, but also were affected by root fly herbivory. Amino acid and sugar concentrations also differed among the populations, but were not affected by root fly herbivory. Overall, population-related differences in plant chemistry were more pronounced for the glucosinolates than for amino acids and sugars. The performance of the root herbivore did not differ among the populations tested. Survival of the root fly was low (<40%), suggesting that other belowground factors may override potential differences in effects related to primary and secondary chemistry.

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.

Susceptibility of Cabbage Maggot Larvae (Diptera: Anthomyiidae) to Hypocreales Entomopathogenic Fungi.

The pathogenicity of six Metarhizium spp., four Beauveria bassiana (Balsamo) Vuillemin, and four Tolypocladium cylindrosporum Gams (Ascomycota: Hypocreales) fungal pathogens exposed to third-instar Delia radicum L. was evaluated in laboratory bioassays. The presence of intra- and intergeneric variations concerning the pathogenicity of the isolates was investigated. Results show that all Metarhizium spp. and T. cylindrosporum isolates caused a noteworthy mortality to the third instar and consequently reduced adult eclosion. The well-known standard, F52 strain (identified as Metarhizium brunneum), resulted in up to 79% reduction in D. radicum eclosion. The other Metarhizium isolates including UAMH 9197 (Metarhizium anisopliae) and UAMH 2801 (M. brunneum), as well as T. cylindrosporum DAOM 167325 and DAOM 183952, produced a mean eclosion reduction of >50%. While the pathogenicity of Metarhizium spp. and T. cylindrosporum is similar, the B. bassiana isolates are undoubtedly less pathogenic. Based on the results obtained with the selected isolates, no intrageneric differences relative to the pathogenicity of the isolates appeared to be present. Globally, this study deepened the knowledge about D. radicum susceptibility toward Hypocreales entomopathogenic fungi, chiefly T. cylindrosporum. The implications of this study regarding the development of a biological control agent are discussed.

Direct plantlet inoculation with soil or insect-associated fungi may control cabbage root fly maggots.

A potential Delia radicum biological control strategy involving cauliflower plantlet inoculation with various fungi was investigated in a series of laboratory and glasshouse experiments. In addition to entomopathogenic fungi, fungi with a high rhizosphere competence and fungi with the ability to survive as saprotrophs in soil were tested. The following fungal species were evaluated in the experiments: Trichoderma atroviride, T. koningiopsis, T. gamsii, Beauveria bassiana, Metharhizium anisopliae, M. brunneum and Clonostachys solani. A commercial carbosulfan-based insecticide was used as a positive control. Additionally, two commercial products, one based on B. bassiana (Naturalis) and one on Bacillus thuringiensis (Delfin) were used as reference biocontrol agents. The aims were (i) to assess the pathogenicity of the selected fungal isolates to Delia radicum, (ii) to evaluate the fungal isolates' rhizosphere competence, with the emphasis on the persistence of the original inoculum on the growing roots, (iii) to assess possible endophytic plant tissue colonization, and (iv) to evaluate potential plant growth stimulating effects of the added inoculi. Significant pathogenicity of tested fungi against Delia radicum was confirmed in in vitro and glasshouse experiments. All tested fungi persisted on cauliflower rhizoplane. More importantly, the added fungi were found on thoroughly washed roots outside the original point of inoculation. This provided us with evidence that our tested fungi could be transferred via or grow with the elongating roots. In addition to colonizing the rhizoplane, some fungi were found inside the plant root or stem tissue, thus exhibiting endophytic characteristics. The importance of fungal ecology as a criterion in appropriate biological control agent selection is discussed.

Effects of regional crop rotations on autumn insect pests in winter oilseed rape.

BACKGROUND: Chemical control of insect pests in oilseed rape (OSR) is becoming increasingly difficult due to the development of resistance and restrictive insecticide approvals in Europe. At the same time, there is a lack of preventive and alternative control measures. Crop rotation mostly fails to control insects due to their mobility; however, changing regional cropping densities can dilute or concentrate pest pressure. In this study, we investigated whether the local occurrence of Psylliodes chrysocephala and Delia radicum, serious insect pests in winter OSR, is influenced by distance from the previous year's OSR fields and how changes in OSR rape cropping density at a regional scale (up to 10 km radius) affect pest pressure. RESULTS: Abundance of P. chrysocephala in yellow water traps decreased with increasing distance to previous year's OSR. Estimated catches in the first 3 weeks of migration were about 68-76% lower at 10 km distance compared to 1 km in autumn 2019 and 2020. However, in both seasons P. chrysocephala was able to disperse over distances of 10 km. Probability of root damage by D. radicum was affected by changes of OSR cropping area at a spatial scale of 2.5 km radius; it increased if acreage of OSR decreased. Furthermore, aphid infestation was lower when OSR was distant in the previous year. CONCLUSION: This study could enable field-specific risk assessment and prediction of pest pressure. To decide about the effectiveness of cropping breaks at a regional level as a preventive crop protection measure, more knowledge on other pest species and antagonists is needed. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Relative Susceptibility of Brassicas to Cabbage Maggot (Diptera: Anthomyiidae) Infestation.

Cabbage maggot, Delia radicum (L.) (Diptera: Anthomyiidae) is a serious pest of Brassica such as broccoli (Brassica oleracea var. italica Plenck) and cauliflower (B. oleracea L. var. botrytis) in California's Central Coast. Since there are limited non-chemical options available for growers to manage D. radicum, there is an urgent need to develop alternative tactics. The objective of this study was to determine the effects of side-by-side plantings of turnip (Brassica rapa var. rapa L.), lettuce (Lactuca sativa L.), cauliflower, and cabbage (B. oleracea L. var. capitata) with broccoli on D. radicum infestation. In 2013 and 2014, the experiments were conducted in Salinas, California. Significantly greater numbers of eggs and larval feeding damage were found on turnip compared with broccoli. Lettuce (Asteraceae), a non-Brassica crop, was compared with broccoli; however, lettuce did not reduce oviposition or larval feeding damage on broccoli. The larval feeding damage on cauliflower was significantly lower than on broccoli when planted side-by-side. The effects on cabbage were not significantly different from broccoli in terms of oviposition and larval feeding damage. This new information generated from the Central Coast of California will be further utilized to develop a trap crop to effectively tackle the D. radicum problem in Brassica fields.

Sex- and maturity-dependent antennal detection of host plant volatiles in the cabbage root fly, Delia radicum.

Adult insect behaviour in response to plant-emitted volatile compounds varies between the sexes and as a function of maturity. These differences in behavioural responses can be due to modulation in the peripheral or central nervous system. In the cabbage root fly, Delia radicum, behavioural effects of certain host plant volatiles on mature female behaviour have been evaluated, and a large number of compounds emitted by brassicaceous host plants have been identified. We recorded here dose-dependent electroantennogram responses to all tested compounds and investigated if the antennal detection of individual volatile compounds emitted by intact and damaged host plants differs between male and female, as well as immature and mature flies. Our results showed dose-dependent responses in mature and immature males and females. Mean response amplitudes varied significantly between sexes for three compounds, and between maturity states for six compounds. For some additional compounds significant differences occurred only for high stimulus doses (interaction between dose and sex and/or dose and maturity status). Multivariate analysis revealed a significant global effect of maturity on electroantennogram response amplitudes and for one experimental session also a significant global effect of the sex. Interestingly, allyl isothiocyanate, a compound stimulating oviposition behaviour, elicited stronger responses in mature than in immature flies, whereas ethylacetophenone, an attractive flower volatile, elicited stronger responses in immature than in mature flies, which correlates with the behavioural role of these compounds. Several host-derived compounds elicited stronger responses in females than in males and, at least at high doses, stronger responses in mature than in immature flies, indicating differential antennal sensitivity to behaviourally active compounds. Six compounds did not cause any significant differences in responses between the different groups of flies. Our results thus confirm peripheral plasticity in plant volatile detection in the cabbage root fly and provide a basis for future behavioural investigations on the function of individual plant compounds.

Diverse cropping systems lead to higher larval mortality of the cabbage root fly (Delia radicum).

Root herbivores pose a major threat to agricultural crops. They are difficult to control and their damage often goes unnoticed until the larvae reach their most devastating late instar stages. Crop diversification can reduce pest pressure, generally without compromising yield. We studied how different diversified cropping systems affected the oviposition and abundance of the specialist cabbage root fly Delia radicum, the most important root herbivore in Brassica crops. The cropping systems included a monoculture, pixel cropping, and four variations of strip cropping with varying intra- and interspecific crop diversity, fertilization and spatial configuration. Furthermore, we assessed whether there was a link between D. radicum and other macroinvertebrates associated with the same plants. Cabbage root fly oviposition was higher in strip cropping designs compared to the monoculture and was highest in the most diversified strip cropping design. Despite the large number of eggs, there were no consistent differences in the number of larvae and pupae between the cropping systems, indicative of high mortality of D. radicum eggs and early instars especially in the strip cropping designs. D. radicum larval and pupal abundance positively correlated with soil-dwelling predators and detritivores and negatively correlated with other belowground herbivores. We found no correlations between the presence of aboveground insect herbivores and the number of D. radicum on the roots. Our findings indicate that root herbivore presence is determined by a complex interplay of many factors, spatial configuration of host plants, and other organisms residing near the roots. Supplementary Information: The online version contains supplementary material available at 10.1007/s10340-023-01629-1.

Root Colonization by Fungal Entomopathogen Systemically Primes Belowground Plant Defense against Cabbage Root Fly.

Entomopathogenic fungi infect insects via spores but also live inside plant tissues as endophytes. Frequently, colonization by entomopathogens provides plants with increased resistance against insects, but the mechanisms are little understood. This study investigated direct, local, and systemic root-mediated interactions between isolates of the fungus Metarhizium brunneum and larvae of the cabbage root fly (CRF) Delia radicum attacking Brassica napus plants. All fungal isolates infected CRF when conidia were present in the soil, leading to 43-93% mortality. Locally, root-associated M. brunneum isolates reduced herbivore damage by 10-20% and in three out of five isolates caused significant insect mortality due to plant-mediated and/or direct effects. A split-root experiment with isolate Gd12 also demonstrated systemic plant resistance with significantly reduced root collar damage by CRF. LC-MS analyses showed that fungal root colonization did not induce changes in phytohormones, while herbivory increased jasmonic acid (JA) and glucosinolate concentrations. Proteinase inhibitor gene expression was also increased. Fungal colonization, however, primed herbivore-induced JA and the expression of the JA-responsive plant defensin 1.2 (PDF1.2) gene. We conclude that root-associated M. brunneum benefits plant health through multiple mechanisms, such as the direct infection of insects, as well as the local and systemic priming of the JA pathway.

Influential Insider: Wolbachia, an Intracellular Symbiont, Manipulates Bacterial Diversity in Its Insect Host.

Facultative intracellular symbionts like the α-proteobacteria Wolbachia influence their insect host phenotype but little is known about how much they affect their host microbiota. Here, we quantified the impact of Wolbachia infection on the bacterial community of the cabbage root fly Delia radicum by comparing the microbiota of Wolbachia-free and infected adult flies of both sexes. We used high-throughput DNA sequencing (Illumina MiSeq, 16S rRNA, V5-V7 region) and performed a community and a network analysis. In both sexes, Wolbachia infection significantly decreased the diversity of D. radicum bacterial communities and modified their structure and composition by reducing abundance in some taxa but increasing it in others. Infection by Wolbachia was negatively correlated to 8 bacteria genera (Erwinia was the most impacted), and positively correlated to Providencia and Serratia. We suggest that Wolbachia might antagonize Erwinia for being entomopathogenic (and potentially intracellular), but would favor Providencia and Serratia because they might protect the host against chemical plant defenses. Although they might seem prisoners in a cell, endocellular symbionts can impact the whole microbiota of their host, hence its extended phenotype, which provides them with a way to interact with the outside world.

Long-lasting effects of antibiotics on bacterial communities of adult flies.

Insect symbionts benefit their host and their study requires large spectrum antibiotic use like tetracycline to weaken or suppress symbiotic communities. While antibiotics have a negative impact on insect fitness, little is known about antibiotic effects on insect microbial communities and how long they last. We characterized the bacterial communities of adult cabbage root fly Delia radicum in a Wolbachia-free population and evaluated the effect of tetracycline treatment on these communities over several generations. Three D. radicum generations were used: the first- and second-generation flies either ingested tetracycline or not, while the third-generation flies were untreated but differed with their parents and/or grandparents that had or had not been treated. Fly bacterial communities were sequenced using a 16S rRNA gene. Tetracycline decreased fly bacterial diversity and induced modifications in both bacterial abundance and relative frequencies, still visible on untreated offspring whose parents and/or grandparents had been treated, therefore demonstrating long-lasting transgenerational effects on animal microbiomes after antibiotic treatment. Flies with an antibiotic history shared bacterial genera, potentially tetracycline resistant and heritable. Next, the transmission should be investigated by comparing several insect development stages and plant compartments to assess vertical and horizontal transmissions of D. radicum bacterial communities.