The cultivation of perennial wild plant mixtures (WPMs) in biogas cropping systems dominated by maize (Zea mays L.) restores numerous ecosystem functions and improves both spatial and temporal agrobiodiversity. In addition, the colorful appearance of WPM can help enhance landscape beauty. However, their methane yield per hectare (MYH) varies greatly and amounts to only about 50% that of maize. This study aimed at decreasing MYH variability and increasing accumulated MYH of WPM by optimizing the establishment method. A field trial was established in southwest Germany in 2014, and is still running. It tested the effects of three WPM establishment procedures (E1: alone [without maize, in May], E2: undersown in cover crop maize [in May], E3: WPM sown after whole‐crop harvest of spring barley [Hordeum vulgare L.] in June) on both MYH and species diversity of two WPMs [S1, S2]). Mono‐cropped maize and cup plant (Silphium perfoliatum L.) were used as reference crops. Of the WPM treatments tested, S2E2 achieved the highest (19,296 normalmnormalN3/ha, 60.5% of maize) and S1E1 the lowest accumulated MYH (8,156 normalmnormalN3/ha, 25.6% of maize) in the years 2014–2018. Cup plant yielded slightly higher than S2E2 (19,968 normalmnormalN3/ha, 62.6% of maize). In 2014, the WPM sown under maize did not significantly affect the cover crop performance. From 2015 onward, E1 and E2 had comparable average annual MYH and average annual number of WPM species. With a similar accumulated MYH but significantly higher number of species (3.5–10.2), WPM S2E2 outperformed cup plant. Overall, the long‐term MYH performance of WPM cultivation for biogas production can be significantly improved by undersowing with maize as cover crop. This improved establishment method could help facilitate the implementation of WPM cultivation for biogas production and thus reduce the trade‐off between bioenergy and biodiversity.

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The global demand for plant biomass to provide bioenergy and heat is continuously increasing because of a growing interest among many industrialized and developing countries towards climate sound and renewable energy supply. The exacerbation of land-use conflicts proliferates social-ecological demands on future bioenergy cropping systems. Perennial herbaceous wild plant mixtures (WPMs) represent an approach to providing social-ecologically more sustainably produced biogas substrate that has gained increasing public and political interest only in recent years. The focus of this study lies on three perennial wild plant species (WPS) that usually dominate the biomass yield performance of WPM cultivation. These WPS were compared with established biogas crops in terms of their substrate-specific methane yield (SMY) and lignocellulosic composition. The plant samples were investigated in a small-scale mesophilic discontinuous biogas batch test for determining the SMY. All WPS were found to have significantly lower SMY (241.5–248.5 lN kgVS−1) than maize (337.5 lN kgVS−1). This was attributed to higher contents of lignin (9.7–12.8% of dry matter) as well as lower contents of hemicellulose (9.9–11.5% of dry matter) in the WPS. Only minor, non-significant differences to cup plant and Virginia mallow were observed. Thus, when planning WPS as a diversification measure in biogas cropping systems, their lower SMY should be considered.