Perennial leafy grasses and legumes are promising for extraction of proteins with a green biorefinery amid increasing demand for local production. However, which defoliation strategy increases radiation-use efficiency and biomass production still needs to be determined. A multi-year field experiment started in 2019 in Denmark with fertilized grasses (perennial ryegrass, tall fescue), unfertilized legumes (alfalfa, red clover) and their fertilized mixture (grass-legume), each defoliated at 2-, 4- or 8- weeks interval, at either 7-9 or 12-14 cm defoliation height. The main aim was to quantify effects of species, defoliation height and frequency, and nitrogen application rate on canopy radiation interception, aboveground biomass and crude protein production, as well as aboveground radiation use efficiency (RUEA).The results showed that accumulated intercepted photosynthetically active radiation (AIpar) increased with increasing defoliation height and decreasing frequency for most systems, with the largest AIpar obtained seasonally by red clover (730-815 MJ m−2) and grass-legume mixture (790-826 MJ m−2) defoliated at medium to low frequency. Tall fescue defoliated at medium to low frequency yielded the highest aboveground biomass (10-14 Mg ha−1) and red clover defoliated at medium to high frequency yielded the most crude protein (2.5-2.9 Mg ha−1) compared to all other systems annually. RUEA decreased with increasing defoliation height and maximum values were achieved at medium frequency, with tall fescue having significantly higher RUEA (1.6-2.1 g MJ−1) than the other systems. Seasonally, RUEA was variable at high defoliation frequency, being consistently higher at the start and the end of the season for ryegrass and the mixture systems.This study pinpoints tall fescue and red clover as perennial crops with high agronomic productivity among tested species for green biorefinery and also concluded that red clover in temperate humid climate maximize productivity and sustainability for biorefining with defoliation at low height and medium frequency.
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