Urinary VOCs as biomarkers of early stage lung tumour development in mice.

BACKGROUND: Lung cancer is the primary cause of cancer-induced death. In addition to prevention and improved treatment, it has increasingly been established that early detection is critical to successful remission. OBJECTIVE: The aim of this study was to identify volatile organic compounds (VOCs) in urine that could help diagnose mouse lung cancer at an early stage of its development. METHODS: We analysed the VOC composition of urine in a genetically engineered lung adenocarcinoma mouse model with oncogenic EGFR doxycycline-inducible lung-specific expression. We compared the urinary VOCs of 10 cancerous mice and 10 healthy mice (controls) before and after doxycycline induction, every two weeks for 12 weeks, until full-blown carcinomas appeared. We used SPME fibres and gas chromatography - mass spectrometry to detect variations in cancer-related urinary VOCs over time. RESULTS: This study allowed us to identify eight diagnostic biomarkers that help discriminate early stages of cancer tumour development (i.e., before MRI imaging techniques could identify it). CONCLUSION: The analysis of mice urinary VOCs have shown that cancer can induce changes in odour profiles at an early stage of cancer development, opening a promising avenue for early diagnosis of lung cancer in other models.

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.

Age-specific allocation of glucosinolates within plant reproductive tissues.

The Optimal Defense Theory (ODT) predicts that the distribution of defenses within a plant should mirror the value and vulnerability of each tissue. Although the ODT has received much experimental support, very few studies have examined defense allocation among reproductive tissues and none assessed simultaneously how these defenses evolve with age. We quantified glucosinolates in perianths, anthers and pistils at different bud maturity stages (i.e., intermediate flower buds, old flower buds and flowers) of undamaged and mechanically damaged plants of an annual brassicaceous species. The youngest leaf was used as a reference for vegetative organs, since it is predicted to be one of the most defended. In line with ODT predictions, reproductive tissues were more defended than vegetative tissues constitutively, and within the former, pistils and anthers more defended than perianths. No change in the overall defense level was found between bud maturity stages, but a significant temporal shift was observed between pistils and anthers. Contrary to ODT predictions, mechanical damage did not induce systemic defenses in leaves but only in pistils. Our results show that defense allocation in plant reproductive tissues occurs at fine spatial and temporal scales, extending the application framework of the ODT. They also demonstrate interactions between space and time in fine-scale defense allocation.

Understanding crop colonization of oilseed rape crops by the cabbage stem flea beetle (Psylliodes chrysocephala L. (Coleoptera: Chrysomelidae)).

BACKGROUND: Development of accurate pest monitoring systems is essential for the establishment of integrated pest management strategies. Information about the pest behavior during the colonization process, as well as the sex and reproductive status of the colonizing population often are lacking and hinder their development. The cabbage stem flea beetle (CSFB, Psylliodes chrysocephala) can cause the complete destruction of oilseed rape crops (OSR, Brassica napus). In the present study, the colonization process of OSR fields by the CSFB was studied. RESULTS: More individuals were caught on the outward facing side of the traps than the side of the trap facing towards the crop at the field border and catches were higher on the trapping units at the center of the field than at its border, suggesting that more beetles were entering than leaving the crop. Catches were higher on lower traps placed near to the crop than on those positioned further from the ground and also were higher during the day than late afternoon and night. The sex-ratio of individuals caught was skewed towards males and sexual maturity was acquired for females during the experiment. Integration of sampling data with local meteorological data showed that the catches correlated mostly with air temperature and relative humidity. CONCLUSION: This study provides new information about the dispersion of the CSFB in OSR fields during the colonization process, and highlights correlations between local meteorological factors and activity of the CSFB, and represent a new step towards implementing monitoring strategies against this pest. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A protocol for increased throughput phenotyping of plant resistance to the pollen beetle.

BACKGROUND: Improving crop resistance to insect herbivores is a major research objective in breeding programs. Although genomic technologies have increased the speed at which large populations can be genotyped, breeding programs still suffer from phenotyping constraints. The pollen beetle (Brassicogethes aeneus) is a major pest of oilseed rape for which no resistant cultivar is available to date, but previous studies have highlighted the potential of white mustard as a source of resistance and introgression of this resistance appears to be a promising strategy. Here we present a phenotyping protocol allowing mid-throughput (i.e., increased throughput compared to current methods) acquisition of resistance data, which could then be used for genetic mapping of QTLs. RESULTS: Contrasted white mustard genotypes were selected from an initial field screening and then evaluated for their resistance under controlled conditions using a standard phenotyping method on entire plants. We then upgraded this protocol for mid-throughput phenotyping, by testing two alternative methods. We found that phenotyping on detached buds did not provide the same resistance contrasts as observed with the standard protocol, in contrast to the phenotyping protocol with miniaturized plants. This protocol was then tested on a large panel composed of hundreds of plants. A significant variation in resistance among genotypes was observed, which validates the large-scale application of this new phenotyping protocol. CONCLUSION: The combination of this mid-throughput phenotyping protocol and white mustard as a source of resistance against the pollen beetle offers a promising avenue for breeding programs aiming to improve oilseed rape resistance. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Odours of cancerous mouse congeners: detection and attractiveness.

Chemical communication plays a major role in social interactions. Cancer, by inducing changes in body odours, may alter interactions between individuals. In the framework of research targeting non-invasive methods to detect early stages of cancer development, this study asked whether untrained mice could detect odour changes in cancerous congeners. If yes, were they able to detect cancer at an early developmental stage? Did it influence female preference? Did variations in volatile organic components of the odour source paralleled mice behavioural responses? We used transgenic mice strains developing or not lung cancer upon antibiotic ingestion. We sampled soiled bedding of cancerous mice (CC) and not cancerous mice (NC), at three experimental conditions: before (T0), early stage (T2) and late stage (T12) of cancer development. Habituation/generalisation and two-way preference tests were performed where soiled beddings of CC and NC mice were presented to wild-derived mice. The composition and relative concentration of volatile organic components (VOC) in the two stimuli types were analysed. Females did not show directional preference at any of the experimental conditions, suggesting that cancer did not influence their choice behaviour. Males did not discriminate between CC and NC stimuli at T0 but did so at T2 and T12, indicating that wild-derived mice could detect cancer at an early stage of development. Finally, although the VOC bouquet differed between CC and NC it did not seem to parallel the observed behavioural response suggesting that other types of odorant components might be involved in behavioural discrimination between CC and NC mice.

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.

Chemical detection triggers honey bee defense against a destructive parasitic threat.

Invasive species events related to globalization are increasing, resulting in parasitic outbreaks. Understanding of host defense mechanisms is needed to predict and mitigate against the consequences of parasite invasion. Using the honey bee Apis mellifera and the mite Varroa destructor, as a host-parasite model, we provide a comprehensive study of a mechanism of parasite detection that triggers a behavioral defense associated with social immunity. Six Varroa-parasitization-specific (VPS) compounds are identified that (1) trigger Varroa-sensitive hygiene (VSH, bees' key defense against Varroa sp.), (2) enable the selective recognition of a parasitized brood and (3) induce responses that mimic intrinsic VSH activity in bee colonies. We also show that individuals engaged in VSH exhibit a unique ability to discriminate VPS compounds from healthy brood signals. These findings enhance our understanding of a critical mechanism of host defense against parasites, and have the potential to apply the integration of pest management in the beekeeping sector.

Impact of Seasonal and Temperature-Dependent Variation in Root Defense Metabolites on Herbivore Preference in Taraxacum officinale.

Plants experience seasonal fluctuations in abiotic and biotic factors such as herbivore attack rates. If and how root defense expression co-varies with seasonal fluctuations in abiotic factors and root herbivore attack rates is not well understood. Here, we evaluated seasonal changes in defensive root latex chemistry of Taraxacum officinale plants in the field and correlated the changes with seasonal fluctuations in abiotic factors and damage potential by Melolontha melolontha, a major natural enemy of T. officinale. We then explored the causality and consequences of these relationships under controlled conditions. The concentration of the defensive sesquiterpene lactone taraxinic acid ß-D glucopyranosyl ester (TA-G) varied substantially over the year and was most strongly correlated to mean monthly temperature. Both temperature and TA-G levels were correlated with annual fluctuations in potential M. melolontha damage. Under controlled conditions, plants grown under high temperature produced more TA-G and were less attractive for M. melolontha. However, temperature-dependent M. melolontha feeding preferences were not significantly altered in TA-G deficient transgenic lines. Our results suggest that fluctuations in temperature leads to variation in the production of a root defensive metabolites that co-varies with expected attack of a major root herbivore. Temperature-dependent herbivore preference, however, is likely to be modulated by other phenotypic alterations.

Plant defense resistance in natural enemies of a specialist insect herbivore.

Plants defend themselves against herbivores through the production of toxic and deterrent metabolites. Adapted herbivores can tolerate and sometimes sequester these metabolites, allowing them to feed on defended plants and become toxic to their own enemies. Can herbivore natural enemies overcome sequestered plant defense metabolites to prey on adapted herbivores? To address this question, we studied how entomopathogenic nematodes cope with benzoxazinoid defense metabolites that are produced by grasses and sequestered by a specialist maize herbivore, the western corn rootworm. We find that nematodes from US maize fields in regions in which the western corn rootworm was present over the last 50 y are behaviorally and metabolically resistant to sequestered benzoxazinoids and more infective toward the western corn rootworm than nematodes from other parts of the world. Exposure of a benzoxazinoid-susceptible nematode strain to the western corn rootworm for 5 generations results in higher behavioral and metabolic resistance and benzoxazinoid-dependent infectivity toward the western corn rootworm. Thus, herbivores that are exposed to a plant defense sequestering herbivore can evolve both behavioral and metabolic resistance to plant defense metabolites, and these traits are associated with higher infectivity toward a defense sequestering herbivore. We conclude that plant defense metabolites that are transferred through adapted herbivores may result in the evolution of resistance in herbivore natural enemies. Our study also identifies plant defense resistance as a potential target for the improvement of biological control agents.

Distinct defense strategies allow different grassland species to cope with root herbivore attack.

Root-feeding insect herbivores are of substantial evolutionary, ecological and economical importance. Plants defend themselves against insect herbivores through a variety of tolerance and resistance strategies. To date, few studies have systematically assessed the prevalence and importance of these strategies for root-herbivore interactions across different plant species. Here, we characterize the defense strategies used by three different grassland species to cope with a generalist root herbivore, the larvae of the European cockchafer Melolontha melolontha. Our results reveal that the different plant species rely on distinct sets of defense strategies. The spotted knapweed (Centaurea stoebe) resists attack by dissuading the larvae through the release of repellent chemicals. White clover (Trifolium repens) does not repel the herbivore, but reduces feeding, most likely through structural defenses and low nutritional quality. Finally, the common dandelion (Taraxacum officinale) allows M. melolontha to feed abundantly but compensates for tissue loss through induced regrowth. Thus, three co-occurring plant species have evolved different solutions to defend themselves against attack by a generalist root herbivore. The different root defense strategies may reflect distinct defense syndromes.

Multivariate Analysis of Multiple Datasets: a Practical Guide for Chemical Ecology.

Chemical ecology has strong links with metabolomics, the large-scale study of all metabolites detectable in a biological sample. Consequently, chemical ecologists are often challenged by the statistical analyses of such large datasets. This holds especially true when the purpose is to integrate multiple datasets to obtain a holistic view and a better understanding of a biological system under study. The present article provides a comprehensive resource to analyze such complex datasets using multivariate methods. It starts from the necessary pre-treatment of data including data transformations and distance calculations, to the application of both gold standard and novel multivariate methods for the integration of different omics data. We illustrate the process of analysis along with detailed results interpretations for six issues representative of the different types of biological questions encountered by chemical ecologists. We provide the necessary knowledge and tools with reproducible R codes and chemical-ecological datasets to practice and teach multivariate methods.

Effect of Planting Depth and Irrigation Regimes on Potato Tuberworm (Lepidoptera: Gelechiidae) Damage Under Central Pivot Irrigation in the Lower Columbia Basin.

Phthorimaea operculella Zeller has proven to be a limitation to potato production. Although pesticides can effectively reduce P. operculella populations, information regarding effective cultural controls is lacking. The following study was designed to evaluate the effect of different irrigation regimes and planting depths to reduce P. operculella damage under central pivot irrigation. Field trials were conducted in 2007 and 2008 and experimental plots followed standard growers' agronomic practices. Three levels of irrigation were used: optimal, 10% less than optimal, and 10% above optimal; potato planting depth was arranged as shallow, normal, and deep; normal planting was considered tubers planted at 0.15 m depth. Mines and number of larvae per plant were counted weekly until harvest and tuber damage, above and below ground, was estimated. Overall, irrigation had a stronger effect than planting depth but there were not a clear planting depth:irrigation interaction. Both years, sampling dates had a strong effect on the number of mines per plant where the number of mines per plant increased as season progressed. There were not clear trends related to mines per plant, mines per tuber or yield related to planting depth, but there were strong tendencies related to irrigation. The data presented herein provide valuable information regarding the potential to adjust irrigation to warrant better P. operculella control.

A mechanism for sequence specificity in plant-mediated interactions between herbivores.

Herbivore communities are shaped by indirect plant-mediated interactions whose outcomes are strongly dependent on the sequence of herbivore arrival. However, the mechanisms underlying sequence specificity are poorly understood. We examined the mechanisms that govern sequence-specific effects of the interaction between two specialist maize herbivores, the leaf feeder Spodoptera frugiperda and the root feeder Diabrotica virgifera virgifera. In the field, S. frugiperda reduces D. v. virgifera abundance, but only when it arrives on the plant first. In behavioral experiments, D. v. virgifera larvae continued feeding on plants that they had infested before leaf infestation, but refused to initiate feeding on plants that were infested by S. frugiperda before their arrival. Changes in root-emitted volatiles were sufficient to elicit this sequence-specific behavior. Root volatile and headspace mixing experiments showed that early-arriving D. v. virgifera larvae suppressed S. frugiperda-induced volatile repellents, which led to the maintenance of host attractiveness to D. v. virgifera. Our study provides a physiological and behavioral mechanism for sequence specificity in plant-mediated interactions and suggests that physiological canalization of behaviorally active metabolites can drive sequence specificity and result in strongly diverging herbivore distribution patterns.

Benefits of jasmonate-dependent defenses against vertebrate herbivores in nature.

Endogenous jasmonates are important regulators of plant defenses. If and how they enable plants to maintain their reproductive output when facing community-level herbivory under natural conditions, however, remains unknown. We demonstrate that jasmonate-deficient Nicotiana attenuata plants suffer more damage by arthropod and vertebrate herbivores than jasmonate-producing plants in nature. However, only damage by vertebrate herbivores translates into a significant reduction in flower production. Vertebrate stem peeling has the strongest negative impact on plant flower production. Stems are defended by jasmonate-dependent nicotine, and the native cottontail rabbit Sylvilagus nuttallii avoids jasmonate-producing N. attenuata shoots because of their high levels of nicotine. Thus, endogenous jasmonates enable plants to resist different types of herbivores in nature, and jasmonate-dependent defenses are important for plants to maintain their reproductive potential when facing vertebrate herbivory. Ecological and evolutionary models on plant defense signaling should aim at integrating arthropod and vertebrate herbivory at the community level.

Influence of environmental parameters on movements and habitat utilization of humpback whales (Megaptera novaeangliae) in the Madagascar breeding ground.

Assessing the movement patterns and key habitat features of breeding humpback whales is a prerequisite for the conservation management of this philopatric species. To investigate the interactions between humpback whale movements and environmental conditions off Madagascar, we deployed 25 satellite tags in the northeast and southwest coast of Madagascar. For each recorded position, we collated estimates of environmental variables and computed two behavioural metrics: behavioural state of 'transiting' (consistent/directional) versus 'localized' (variable/non-directional), and active swimming speed (i.e. speed relative to the current). On coastal habitats (i.e. bathymetry < 200 m and in adjacent areas), females showed localized behaviour in deep waters (191 ± 20 m) and at large distances (14 ± 0.6 km) from shore, suggesting that their breeding habitat extends beyond the shallowest waters available close to the coastline. Males' active swimming speed decreased in shallow waters, but environmental parameters did not influence their likelihood to exhibit localized movements, which was probably dominated by social factors instead. In oceanic habitats, both males and females showed localized behaviours in shallow waters and favoured high chlorophyll-a concentrations. Active swimming speed accounts for a large proportion of observed movement speed; however, breeding humpback whales probably exploit prevailing ocean currents to maximize displacement. This study provides evidence that coastal areas, generally subject to strong human pressure, remain the core habitat of humpback whales off Madagascar. Our results expand the knowledge of humpback whale habitat use in oceanic habitat and response to variability of environmental factors such as oceanic current and chlorophyll level.

Manipulating feeding stimulation to protect crops against insect pests?

Enhancing natural mechanisms of plant defense against herbivores is one of the possible strategies to protect cultivated species against insect pests. Host plant feeding stimulation, which results from phagostimulant and phagodeterrent effects of both primary and secondary metabolites, could play a key role in levels of damage caused to crop plants. We tested this hypothesis by comparing the feeding intensity of the pollen beetle Meligethes aeneus on six oilseed rape (Brassica napus) genotypes in a feeding experiment, and by assessing the content of possible phagostimulant and phagodeterrent compounds in tissues targeted by the insect (flower buds). For this purpose, several dozens of primary and secondary metabolites were quantified by a set of chromatographic techniques. Intergenotypic variability was found both in the feeding experiment and in the metabolic profile of plant tissues. Biochemical composition of the perianth was in particular highly correlated with insect damage. Only a few compounds explained this correlation, among which was sucrose, known to be highly phagostimulating. Further testing is needed to validate the suggested impact of the specific compounds we have identified. Nevertheless, our results open the way for a crop protection strategy based on artificial selection of key determinants of insect feeding stimulation.

Interplay between parasitism and host ontogenic resistance in the epidemiology of the soil-borne plant pathogen Rhizoctonia solani.

Spread of soil-borne fungal plant pathogens is mainly driven by the amount of resources the pathogen is able to capture and exploit should it behave either as a saprotroph or a parasite. Despite their importance in understanding the fungal spread in agricultural ecosystems, experimental data related to exploitation of infected host plants by the pathogen remain scarce. Using Rhizoctonia solani / Raphanus sativus as a model pathosystem, we have obtained evidence on the link between ontogenic resistance of a tuberizing host and (i) its susceptibility to the pathogen and (ii) after infection, the ability of the fungus to spread in soil. Based on a highly replicable experimental system, we first show that infection success strongly depends on the host phenological stage. The nature of the disease symptoms abruptly changes depending on whether infection occurred before or after host tuberization, switching from damping-off to necrosis respectively. Our investigations also demonstrate that fungal spread in soil still depends on the host phenological stage at the moment of infection. High, medium, or low spread occurred when infection was respectively before, during, or after the tuberization process. Implications for crop protection are discussed.