Integrated Characterisation of Elite Wild Lolium perenne Germplasm Reveals Valuable Parental Material for Sustainable Forage Breeding

The identification of elite forage germplasm combining superior agronomic performance with enhanced nutritional and functional properties is essential for sustainable livestock production. This study evaluated a collection of 34 wild Lolium perenne genotypes collected from three geographically distinct regions of southwestern Romania and maintained under ex situ conditions. Morphological evaluation using hierarchical clustering and principal component analysis (PCA) identified five elite genotypes, which were subsequently characterised for proximate composition, mineral content, total phenolic content, antioxidant activity (DPPH and FRAP), and their ability to modulate probiotic bacterial growth in vitro. Protein content ranged from 132.2 to 163.2 g/kg, while total phenolic content varied between 8.56 and 10.35 mg GAE/g DW. Antioxidant capacity ranged from 17.73 to 22.34 μmol TE/g DW by DPPH and from 28.61 to 30.65 μmol Fe2+/g DW by FRAP. Growth modulation assays revealed marked genotype- and strain-dependent responses, with maximum potentiation coefficients (CPs) of 188.81% for Bifidobacterium lactis BB-12, 225.77% for the mixed BB-12/Lactobacillus acidophilus LA-5 culture, and 349.32% for Lactobacillus casei DG at 1055 µg/mL. Overall, P.14 and P.26 were distinguished by their antioxidant profiles, whereas P.17 showed particular potential in terms of probiotic growth-promoting activity. These findings demonstrate that integrated phenotypic, nutritional, phytochemical, and functional evaluation is an effective strategy for identifying elite wild L. perenne germplasm suitable as parental material for sustainable forage breeding.