Alleviation of gadolinium stress on Medicago by elevated atmospheric CO₂ is mediated by changes in carbohydrates, Anthocyanin, and proline metabolism

Rare earth elements (REE) like Gadolinium (Gd), are increasingly used in industry and agriculture and this is concomitant with the increasingly leaking of Gd into the environment. Under a certain threshold concentration, REE can promote plant growth, however, beyond this concentration, they exert negative effects on plant growth. Moreover, the effect of Gd on plants growth and metabolism under a futuristic climate with increasingly atmospheric CO₂ has not yet been studied. To this end, we investigated the effect of soil contamination with Gd (150 mg/kg soil) on the growth, carbohydrates, proline, and anthocyanin metabolism of Medicago plants grown under ambient (aCO₂, 410 ppm) or elevated CO₂ (eCO₂, 720 ppm) concentration. Gd negatively affected the growth and photosynthesis of plants and imposed oxidative stress i.e., increased H₂O₂ and lipid peroxidation (MDA) level. As defense lines, the level and metabolism of osmoprotectants (soluble sugars and proline) and antioxidants (phenolics, anthocyanins, and tocopherols) were increased under Gd treatment. High CO₂ positively affected the growth and metabolism of Medicago plants. Moreover, eCO₂ mitigated the negative impacts of Gd on Medicago growth. It further induced the levels of osmoprotectants and antioxidants. In line with increased proline and anthocyanins, their metabolic enzymes (e.g. OAT, P5CS, PAL, and CS) were also increased. This study advances our understanding of how Gd adversely affects Medicago plant growth and metabolism. It also sheds light on the biochemical mechanisms underlying the Gd stress-reducing impact of eCO_2.