LEPRI, A., KAZMI, H., BERTOLOTTI, F., LONGO, E., OCCHIGROSSI, S., QUATTROCCHI, L., DE VIVO, M., SCINTU, D., SVOLACCHIA, N., TARKOWSKÁ, Danuše, TUREČKOVÁ, Veronika, STRNAD, Miroslav, DEL BIANCO, M., DI MAMBRO, G., COSTANTINO, P., SABATINI, F., DELLO IOIO, R., VITTORIOSO, P. A DOF transcriptional repressor-gibberellin feedback loop plays a crucial role in modulating light-independent seed germination. Plant Communications. 2025, 6(4), 101262. ISSN 2590-3462. E-ISSN 2590-3462

Plants have evolved several strategies to cope with the ever-changing environment. One example of this is given by seed germination, which must occur when environmental conditions are suitable for plant life. In the model system Arabidopsis thaliana seed germination is induced by light, however, in nature, seeds of several plant species can germinate regardless of this stimulus. While the molecular mechanisms underlying light-induced seed germination are well understood, those governing germination in the dark are still vague, mostly due to the lack of suitable model systems. Here, we employ Cardamine hirsuta, a close relative of Arabidopsis, as a powerful model system to uncover the molecular mechanisms underlying light-independent germination. By comparing Cardamine and Arabidopsis, we show that maintenance of the pro-germination hormone gibberellin (GA) levels prompt Cardamine seeds to germinate under both dark and light conditions. Using genetic and molecular biology experiments, we show that the Cardamine DOF transcriptional repressor DOF AFFECTING GERMINATION 1 (ChDAG1), homologous to the Arabidopsis transcription factor DAG1, is involved in this process functioning to mitigate GA levels by negatively regulating GA biosynthetic genes ChGA3OX1 and ChGA3OX2, independently of light conditions. We also demonstrate that this mechanism is likely conserved in other Brassicaceae species capable of germinating in dark conditions, such as Lepidium sativum and Camelina sativa. Our data support Cardamine as a new model system suitable for studying light-independent germination studies. Exploiting this system, we have also resolved a long-standing question about the mechanisms controlling light-independent germination in plants, opening new frontiers for future research.