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Simple Summary: Carabids are abundant and diverse arthropods in agroecosystems and are well known for their generalist diet and therefore biocontrol potential. Most studies on carabids' diet have focused on periods of peak pest infestations but their feeding habits through the cropping season are poorly known. In this study, we explored how carabids' trophic interactions with pests (aphids and slugs) and alternative prey (spiders, earthworms and springtails) varied from the wheat seedling emergence to wheat ripening by analysing the prey DNA in their gut. Carabids seemed to switch between prey depending on their availability at the time, reaffirming carabids' opportunistic diet. Regarding biocontrol, carabid taxa contributed complementarily to pest predation over the season and per pest taxon, highlighting the importance of favouring diversity in agroecosystems. To date, evaluating the diets of natural enemies like carabids has largely been limited to spatially explicit and short-term sampling. This leaves a knowledge gap for the intra-annual dynamics of carabid diets, and the provision and timing of delivery of natural pest control services. Season-long pitfall trapping of adult carabids was conducted in conventional winter wheat fields, from November 2018 to June 2019, in five French departments. Diagnostic Multiplex PCR of carabid gut contents was used to determine the dynamics of carabid diets. The overall detection rate of target prey DNA was high across carabid individuals (80%) but varied with the prey group. The rate of detection was low for pests, at 8.1% for slugs and 9.6% for aphids. Detection of intraguild predation and predation on decomposers was higher, at 23.8% for spiders, 37.9% for earthworms and 64.6% for springtails. Prey switching was high at the carabid community level, with pest consumption and intraguild predation increasing through the cropping season as the availability of these prey increased in the environment, while the detection of decomposer DNA decreased. Variation in diet through the cropping season was characterized by: (i) complementary predation on slug and aphid pests; and (ii) temporal complementarity in the predominant carabid taxa feeding on each pest. We hypothesize that natural pest control services delivered by carabids are determined by complementary contributions to predation by the different carabid taxa over the season. [ABSTRACT FROM AUTHOR]

Belowground herbivores impact plant performance, thereby inducing changes in plant community composition, which potentially leads to cascading effects onto higher trophic levels and ecosystem processes and productivity. Among soil-living insects, external root-chewing generalist herbivores have the strongest impact on plants. However, the lack of knowledge on their feeding behaviour under field conditions considerably hampers achieving a comprehensive understanding of how they affect plant communities. Here, we address this gap of knowledge by investigating the feeding behaviour of Agriotes click beetle larvae, which are common generalist external root-chewers in temperate grassland soils. Utilizing diagnostic multiplex PCR to assess the larval diet, we examined the seasonal patterns in feeding activity, putative preferences for specific plant taxa, and whether species identity and larval instar affect food choices of the herbivores. Contrary to our hypothesis, most of the larvae were feeding-active throughout the entire vegetation period, indicating that the grassland plants are subjected to constant belowground feeding pressure. Feeding was selective, with members of Plantaginaceae and Asteraceae being preferred; Apiaceae were avoided. Poaceae, although assumed to be most preferred, had an intermediate position. The food preferences exhibited seasonal changes, indicating a fluctuation in plant traits important for wireworm feeding choice. Species- and instar-specific differences in dietary choice of the Agriotes larvae were small, suggesting that species and larval instars occupy the same trophic niche. According to the current findings, the food choice of these larvae is primarily driven by plant identity, exhibiting seasonal changes. This needs to be considered when analysing soil herbivore-plant interactions. [ABSTRACT FROM AUTHOR]

Plant roots represent an important food source for soil-dwelling animals, but tracking herbivore food choices below-ground is difficult. Here, we present an optimized PCR assay for the detection of plant DNA in the guts of invertebrates, using general plant primers targeting the trn T- F chloroplast DNA region. Based on this assay, we assessed the influence of plant identity on the detectability of ingested plant DNA in Agriotes click beetle larvae. Six different plant species were fed to the insects, comprising a grass, a legume and four nonlegume forbs. Moreover, we examined whether it is possible to amplify DNA of decaying plants and if DNA of decayed plant food is detectable in the guts of the larvae. DNA of the ingested roots could be detected in the guts of the larvae for up to 72-h post-feeding, the maximum digestion time tested. When fed with living plants, DNA detection rates differed significantly between the plant species. This may be ascribed to differences in the amount of plant tissue consumed, root palatability, root morphology and/or secondary plant components. These findings indicate that plant identity can affect post-feeding DNA detection success, which needs to be considered for the interpretation of molecularly derived feeding rates on plants. Amplification of plant DNA from decaying plants was possible as long as any tissue could be retrieved from the soil. The consumption of decaying plant tissue could also be verified by our assay, but the insects seemed to prefer fresh roots over decaying plant material. [ABSTRACT FROM AUTHOR]