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Flavonoids play a key role in the regulation of plant–plant and plant–microbe interactions, and factors determining their release have been investigated in most of the common forage legumes. However, little is known about the response of flavonoid production and release to co-cultivation with other crop species. This study investigated alterations in the concentration of flavonoids in plant tissues and root exudates in four legumes [alfalfa ( Medicago sativa L. ), black medic ( Medicago polymorpha L. ), crimson clover ( Trifolium incarnatum L. ), and subterranean clover ( Trifolium subterraneum L. )] co-cultivated with durum wheat [ Triticum turgidum subsp. durum (Desf.) Husn.]. For this purpose, we carried out two experiments in a greenhouse, one with glass beads as growth media for root exudate extraction and one with soil as growth media for flavonoid detection in shoot and root biomass, using LC–MS/MS analysis. This study revealed that interspecific competition with wheat negatively affected legume growth and led to a significant reduction in shoot and root biomass compared with the same legume species grown in monoculture. In contrast, the concentration of flavonoids significantly increased both in legume biomass and in root exudates. Changes in flavonoid concentration involved daidzein, genistein, medicarpin, and formononetin, which have been found to be involved in legume nodulation and regulation of plant–plant interaction. We hypothesize that legumes responded to the co-cultivation with wheat by promoting nodulation and increasing exudation of allelopathic compounds, respectively, to compensate for the lack of nutrients caused by the presence of wheat in the cultivation system and to reduce the competitiveness of neighboring plants. Future studies should elucidate the bioactivity of flavonoid compounds in cereal-legume co-cultivation systems and their specific role in the nodulation process and inter-specific plant interactions such as potential effects on weeds.

Weed control in organic conservative vegetable systems is extremely challenging and the use of legume permanent living mulches (pLM) presents an interesting opportunity. The successful use of pLM is largely determined by the choice of appropriate legumes which are able to combine adequate weed control with a marginal competitive effect on the cash crop(s). However, the availability of legumes for such systems is limited and their characterization based on growth traits can support the selection of suitable legumes for conservation organic vegetable systems. The current study investigated weed control capacity and variability in morphological and phenological traits relevant in inter-plant competition among a range of 11 commercial cultivars of legumes and seven ecotypes of Medicago polymorpha (bur medic). For commercial cultivars, Lotus corniculatus (bird’s-foot trefoil) and Trifolium repens (white clover) showed the best weed control capacity, while Trifolium subterraneum (subterranean clover) and Medicago polymopha had more suitable characteristics for a rapid and complete establishment of the pLM. Overall, legume mulches appear more effective in dicotyledonous than in monocotyledonous weed control. Trifolium subterraneum cv. Antas and T. repens cv. Haifa were identified as the potentially most suitable legumes for use as pLM and their use in mixtures could be a promising solution. In addition, the ecotypes of Medicago polymorpha Manciano and Talamone proved to be well adapted for local environmental conditions and they showed a better weed suppression than the commercial cultivars of Medicago polymorpha.

Relay intercropping of subsidiary legumes with durum wheat (living mulch) can be a viable option to support ecological weed control and optimize nutrient cycling in cereal-based cropping systems. However, the lack of knowledge on suitable legume species is often identified as the main bottleneck for the successful application of legume living mulches. This study aimed to evaluate the suitability of 12 different legumes for relay intercropping with wheat in two contrasting Mediterranean cereal-based cropping systems respectively characterized by low-input and integrated management. Each legume was monitored from the undersowing in wheat until the following spring and we compared direct drilling to broadcast sowing of legumes. None of the undersown legumes showed a negative effect on the wheat grain yield. Relay intercropping of legumes proved to be an effective solution to control weeds before and after the wheat harvest, provided suitable legumes species are chosen. Suitable legumes reduced the weed biomass up to the 90% during the intercropping and up to 94% in the following spring. On the contrary, legumes such as Trifoliumresupinatum, Viciavillosa, Medicagotruncatula, and Medicagoscutellata boosted weed growth in the following spring in comparison with the control. According to the performance of legumes, Medicagosativa, Trifoliumrepens and Medicagolupulina had the most suitable characteristics for relay intercropping with durum wheat at the Ravenna site, in a highly productive region whereas Medicagosativa, Hedysarumcoronarium and Trifoliumsubterraneum performed better in the low-input system near Pisa, where yields are generally lower. This is the first time that such a diversity in legumes species is tested in the same experiment for relay intercropping under diversified environmental and management conditions. The results of this study can support farmers in selecting the most appropriated legume species for their specific cropping systems and local conditions.

Wild edible plant species are often more tolerant to salinity than many crop plants. Considering the salinization of irrigation water that is progressively affecting the Mediterranean region and the market demand for new plant foods, the cultivation of wild edible species could represent a valid alternative to conventional vegetable crops. In this study, Reichardia picroides (L.) Roth, a widespread spontaneous herb of ethnobotanical tradition, was grown for four or six weeks under a greenhouse in a floating system for the production of baby leaves. In order to improve the nutraceutical quality of the tissues, the plants were exposed to the following NaCl concentrations in the nutrient solution: 1.7 (control), 25, 50, and 100 mM. The results showed that a 4-week growing period in a floating system with 50 mM NaCl in the nutrient solution increased the content of bioactive molecules without affecting the fresh yield. After six weeks of cultivation, despite a decrease in biomass production as compared with the control, the leaves of salt-treated plants contained higher levels of bioactive molecules along with lower amounts of nitrate ion.

Relay intercropping is considered a valuable agroecological practice to increase and stabilize crop yields while ensuring the provision of several ecosystem services as well as sustainability and resilience to changing climatic conditions. However, farmers are still reluctant in the use of intercropping practices since there is a huge knowledge gap regarding the time of sowing, sowing ratio, crop stand density, and cultivar choice. In this study, we carried out a 3-year field experiment in Central Italy to assess the effect of relay intercropping on the agronomic performance and competitiveness of winter durum wheat (Triticum durum Desf. cv. Minosse) and spring lentil (Lens culinaris Medik. cv. Elsa) under a low-input management system, comparing different crop stand types (monocrop vs. intercrop) and target plant densities (350 plants m2—full dose vs. 116 plants m2—1/3 dose). The results revealed that intercropping increased grain yield compared to monocropping: significantly (p < 0.0001) against both monocrops in 2021 and non-significantly against durum wheat in 2019 and 2020. Yield advantage in both intercropping systems ranged between 164 and 648%. Durum wheat competitiveness was stronger in 2019 and 2021, while lentil was the most competitive component in 2020. Intercropping favored P accumulation in durum wheat shoots. There was no difference in grain yield of both crops between the highly- and lowly-dense system in 2020 and 2021. Both intercropping strategies were as effective as mechanical hoeing in controlling weeds and proved beneficial in stabilizing lentil productivity. Further economic analysis capturing the additional costs incurred in intercropping and mechanical weeding would highlight the magnitude of profitability of these systems.

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