Cattle production based on natural pastures is often subject to flooding periods, which affect plant performance and as a result, forage production. Although most forage legumes are not tolerant to flooding, Lotus spp. are outstanding alternatives, since species, such as L. tenuis (Lt) and L. corniculatus (LcT), have high forage quality and are adaptable to different environments. We recently obtained a L. tenuis × L. corniculatus hybrid (LtxLc) with potential new cultivar traits, although its tolerance to flooding stress has not yet been evaluated. In the present study, the performance of LtxLc, its parental diploid accessions, the model legume L. japonicus and tetraploid LcT were evaluated under 55 days of partial submergence stress and a 35‐day recovery period. Physiological, morphological and anatomical traits were analysed, showing that tolerance to partial submergence was positively associated with aerenchyma and adventitious root formation and relative growth rates. Overall, Lt and LtxLc showed the best responses under stress and during the recovery period. Nevertheless, the higher forage value of LtxLc makes it recommendable for use in environments affected by flooding. Our results could be used as breeding criteria for the generation of new cultivars tolerant to partial submergence stress.

By exploiting interspecific hybrids and their progeny, we identified key regulatory and transporter genes intimately related to proanthocyanidin biosynthesis in leaves of Lotus spp. Proanthocyanidins (PAs), known as condensed tannins, are polymeric flavonoids enriching forage legumes of key nutritional value to prevent bloating in ruminant animals. Unfortunately, major forage legumes such as alfalfa and clovers lack PAs in edible tissues. Therefore, engineering the PA trait in herbage of forage legumes is paramount to improve both ecological and economical sustainability of cattle production system. Progresses on the understanding of genetic determinants controlling PA biosynthesis and accumulation have been mainly made studying mutants of Arabidopsis, Medicago truncatula and Lotus japonicus, model species unable to synthesize PAs in the leaves. Here, we exploited interspecific hybrids between Lotus corniculatus, with high levels of PAs in the leaves, and Lotus tenuis, with no PAs in these organs, and relative F progeny, to identify among candidate PA regulators and transporters the genes mainly affecting this trait. We found that the levels of leaf PAs significantly correlate with the expression of MATE1, the putative transporter of glycosylated PA monomers, and, among the candidate regulatory genes, with the expression of the MYB genes TT2a, TT2b and MYB14 and the bHLH gene TT8. The expression levels of TT2b and TT8 also correlated with those of all key structural genes of the PA pathways investigated, MATE1 included. Our study unveils a different involvement of the three Lotus TT2 paralogs to the PA trait and highlights differences in the regulation of this trait in our Lotus genotypes with respect to model species. This information opens new avenues for breeding bloat safe forage legumes.