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.

Chromatin accessibility profiling and transcriptomics of Marchantia polymorpha heat‑shock transcription factor (HSF) mutants reveal that HSFA1 governs the placement of cis‑regulatory elements for heat‑induced gene activation, a mechanism conserved across plants, mice, and humans. Integrated gene regulatory network modeling identifies MpWRKY10 and MpABI5B as indirect regulators linking phenylpropanoid and stress pathways, while abscisic acid influences gene expression downstream of HSFA1 without broadly reshaping chromatin. A cross‑species, cross‑condition machine‑learning framework successfully predicts chromatin accessibility and expression, underscoring a conserved regulatory logic in stress responses.