Weed control by crops through growth suppressive root exudates is a promising alternative to herbicides. Buckwheat (Fagopyrum esculentum) is known for its weed suppression and redroot pigweed (Amaranthus retroflexus) control is probably partly due to allelopathic root exudates. This work studies whether other weeds are also suppressed by buckwheat and if the presence of weeds is necessary to induce growth repression. Buckwheat and different weeds were co-cultivated in soil, separating roots by a mesh allowing to study effects due to diffusion. Buckwheat suppressed growth of pigweed, goosefoot and barnyard grass by 53, 42, and 77% respectively without physical root interactions, probably through allelopathic compounds. Root exudates were obtained from sand cultures of buckwheat (BK), pigweed (P), and a buckwheat/pigweed mixed culture (BK-P). BK-P root exudates inhibited pigweed root growth by 49%. Characterization of root exudates by UHPLC-HRMS and principal component analysis revealed that BK and BK-P had a different metabolic profile suggesting that buckwheat changes its root exudation in the presence of pigweed indicating heterospecific recognition. Among the 15 different markers, which were more abundant in BK-P, tryptophan was identified and four others were tentatively identified. Our findings might contribute to the selection of crops with weed suppressive effects.

Within the plant kingdom, there is an exceptional amount of chemical diversity that has yet to be annotated. It is for this reason that non-targeted analysis is of interest for those working in novel natural products. To increase the number and diversity of compounds observable in root exudate extracts, several workflows which differ at three key stages were compared: 1) sample extraction, 2) chromatography, and 3) data preprocessing. Plants were grown in Hoagland's solution for two weeks, and exudates were initially extracted with water, followed by a 24-h regeneration period with subsequent extraction using methanol. Utilizing the second extraction showed improved results with less ion suppression and reduced retention time shifting compared to the first extraction. A single column method, utilizing a pentafluorophenyl column, paired with high-resolution mass spectrometry ionized and correctly identified 34 mock root exudate compounds, while the dual column method, incorporating a pentafluorophenyl column and a porous graphitic carbon column, retained and identified 43 compounds. In a pooled quality control sample of exudate extracts, the single column method detected 1,444 compounds. While the dual method detected fewer compounds overall (1,050), it revealed a larger number of small polar compounds. Three preprocessing methods (targeted, proprietary, and open source) successfully identified 43, 31, and 38 mock root exudate compounds to confidence level 1, respectively. Enhancing signal strength and analytical method stability involves removing the high ionic strength nutrient solution before sampling root exudate extracts. Despite signal intensity loss, a dual column method enhances compound coverage, particularly for small polar metabolites. Open-source software proves a viable alternative for non-targeted analysis, even surpassing proprietary software in peak picking.

This study investigates the effects of substrate composition on root architecture, plant growth, and allelopathic secondary metabolites, specifically benzoxazinoids (BXs), in the rhizospheres of rye (Secale cereale L.) and redroot pigweed (Amaranthus retroflexus L.). Given the complexities of root exudate analysis, including the influence of substrate on root morphology and exudation, the experiment compared plant growth and BX release in two substrates: glass microbeads and a mixture of clay beads and attapulgite. Rye, pigweed, and co-cultures of the two were grown under controlled conditions, with root and shoot parameters measured to assess substrate suitability. Additionally, UPLC-QTOF-MS was used to analyze BXs in rye and rye–pigweed co-cultures. The results demonstrated that the clay bead and attapulgite mixture provided better growth conditions and was effective for BX extraction, making it a suitable substrate for studying allelopathy in controlled environments. The findings highlight the critical role of substrate composition in both plant development and the study of root exudates, with implications for better understanding of crop–weed interactions and allelopathy.

Root exudates are composed of primary and secondary organic compounds that function as signalling molecules and play important roles in plant-environment interactions. The quantity and composition of root exudates vary depending on the species, genotype, and environmental conditions, including the presence and identity of neighbouring plants. Although cover crops are commonly used in agricultural practices for their ecosystem services, such as weed suppression, pathogen control, and soil structure improvement, studies on their root exudates are limited. Our study provides the first characterization of the root exudates of black oat interacting with weed neighbours, redroot pigweed and blackgrass, as well as with neighbours of the same species, black oat. We investigated how these interactions influence the black oat root exudation patterns. Furthermore, we investigated the impact of black oat presence on neighbours and how exposure to black oat root exudate treatments affects the root traits of weeds. The upregulated compounds detected in root exudates in response to neighbouring plants primarily belonged to the organic oxygen compounds superclass, with most of which identified as amino acids and carbohydrates. In the presence of redroot pigweed, a general increase in root exudation was observed, with amino acids and sugar sulphates being upregulated. The presence of black oat had varying effects among the neighbouring plants. While significant decreases observed and black oat in redroot pigweed root traits, an increase was observed in blackgrass. Similarly, more pronounced effects were observed in redroot pigweed compared to blackgrass upon root exudate application. This study provides a characterization and insights into the dynamic nature of root exudates and their influence on root traits and plant interactions.

Agroecological crop protection (ACP) aims to reduce pesticide reliance by redesigning cropping systems to enhance ecological processes for pest control, yet its feasibility and performance under real farming conditions remains insufficiently documented across crops. This study compares agronomic and ecological outcomes of paired agroecological and control fields (following Swiss standard integrated management) across a Swiss network of commercial farms for three major arable crops in a three-year experiment. We quantified the extent of implementation of 24 different agroecological crop protection practices co-designed with farmers, as well pesticide use reduction, and assessed their effects on weeds, diseases and insect pest pressure, natural enemy abundance, and crop yield, while accounting for environmental factors such as soil properties, weather, and landscape composition. Undersowing, flower strips, mechanical weeding, varietal mixture, false seedbed and push-pull were significantly more implemented on the agroecological field than on the control, and pesticide use was strongly reduced. However, results regarding crop protection and performance were strongly crop-dependent. In wheat, yields were maintained despite slightly higher weed biomass. In barley, ACP limited noxious organisms, yet yields declined, suggesting a combination of factors including non-biotic constraints. In oilseed rape, higher weed biomass and pest pressure led to reduced yields, despite increased natural enemy abundance. Landscape composition, weather conditions and soil characteristics contributed to variability of the response parameters. These results show that ACP can substitute for chemical control in some systems, but the effectiveness depends on crop-specific traits and management practices. Overall, successful pesticide reduction requires context-specific, system-based strategies supported by technical expertise.