The study characterizes the single-copy S-nitrosoglutathione reductase 1 (MpGSNOR1) in the liverwort Marchantia polymorpha, showing that loss-of-function mutants generated via CRISPR/Cas9 exhibit marked morphological defects and compromised SNO homeostasis and immune responses. These findings indicate that GSNOR-mediated regulation of S‑nitrosylation is an ancient mechanism linking development and immunity in early land plants.

The authors adapted OpenPlant kit CRISPR/Cas9 tools to enable multiplex gRNA expression from a single transcript using tRNA sequences in the liverwort Marchantia polymorpha, markedly enhancing editing efficiency and scalability. They coupled this vector system with a simplified, optimized thallus transformation protocol, providing a rapid and versatile platform for generating CRISPR/Cas9 mutants and advancing functional genomics in this model species.