CIPK-B is essential for salt stress signalling in Marchantia polymorpha

Abstract

Calcium signalling is central to many plant processes, with families of calcium decoder proteins having expanded across the green lineage and redundancy existing between decoders. The liverwort Marchantia polymorpha has fast become a new model plant, but it is unclear what calcium decoders exist in this species. We have performed phylogenetic analyses to identify the Calcineurin B-Like (CBL) and CBL-Interacting Protein Kinase (CIPK) network of M. polymorpha. We analysed CBL-CIPK expression during salt stress, and determined protein- protein interactions using yeast two-hybrid and bimolecular fluorescence complementation. We also created genetic knockouts using CRISPR/Cas9. We confirm that M. polymorpha has two CIPKs and three CBLs. Both CIPKs and only one CBL show salt-responsive transcriptional changes. All M. polymorpha CBL-CIPKs interact with each other in planta. Knocking out CIPK-B causes increased sensitivity to salt suggesting that this CIPK is involved in salt signalling. We have identified CBL-CIPKs that form part of a salt tolerance pathway in M. polymorpha. Phylogeny and interaction studies imply that these CBL-CIPKs form an evolutionarily conserved Salt Overly Sensitive (SOS) pathway. Hence, salt responses may be some of the early functions of CBL-CIPK networks and increased abiotic stress tolerance required for land plant emergence.