The authors introduced a polycistronic tRNA‑gRNA array for CRISPR/Cas9 editing in Physcomitrium patens that doubled the frequency of large, targeted deletions compared with conventional single‑gRNA constructs. Using dual‑gRNA targeting, they achieved simultaneous deletion of two to four genes (katanin and TPX2 families) in a single transformation, reaching up to 42% efficiency per gene, though efficiency depended on gRNA pair design.

The study identifies the Arabinogalactan protein SLEEPING BEAUTY (SB) from the bryophyte Physcomitrium patens as a highly glycosylated, intrinsically disordered protein that rigidifies the cell wall and facilitates orderly cellulose microfibril deposition, with loss of function leading to softened walls. Critical proline‑linked glycosylation sites required for SB’s disordered conformation, osmotic stress‑induced endocytosis, and tonoplast trafficking were pinpointed using pharmacological inhibition and mutagenesis, and the wall‑rigidifying role of SB was shown to be conserved in Arabidopsis.

The study characterizes telomere repeat binding (TRB) proteins in the model moss Physcomitrium patens, demonstrating that individual PpTRB genes are essential for normal protonemal and gametophore development and that loss of TRBs leads to telomere shortening, mirroring findings in seed plants. Transcriptome analysis of TRB mutants shows altered expression of genes linked to transcription regulation and stimulus response, while subcellular localization confirms nuclear residence and mutual interaction of PpTRBs, underscoring their conserved role in telomere maintenance across land plants.

The study reveals that MAX2-dependent signaling suppresses cytokinin accumulation and thus delays gametophore bud formation in the moss Physcomitrium patens, while loss of SMXL repressors accelerates the transition from 2D protonema to 3D shoot growth. Cytokinin levels were elevated in Ppmax2 mutants, and exogenous cytokinin rescued the delayed bud formation of SMXL mutants, indicating a sequential hormonal control involving the unidentified KAI2 ligand and cytokinin.

The study identifies a moss‑specific fusion protein, Rosetta NATD‑MAPK 1 (RAK1), that combines a MAPK domain with an N‑acetyltransferase and demonstrates that its acetyltransferase activity is enhanced upon MAPK activation. Knockout of RAK1 impairs the 2D‑to‑3D developmental transition in Physcomitrium patens, and mass‑spectrometry reveals associated changes in acetylation and phosphorylation linked to metabolic reprogramming.

The study uncovers that the sole Flotillin protein in the moss Physcomitrium patens localizes to chloroplast thylakoid membranes and influences stress responses. Overexpression of PpFLOT reduces salinity tolerance, triggers necrotic phenotypes in liquid culture, and alters chlorophyll, H₂O₂, pigment, and lipid profiles, implicating the protein in Ca²⁺ and ROS signaling during abiotic and biotic stress.

The study characterizes the distinct and overlapping roles of the rice PI paralogs OsMADS2 and OsMADS4 in lodicule specification, flowering time, and floral organ development by analyzing null and double mutants and overexpression lines. Genome-wide binding (ChIP‑seq) and transcriptome (RNA‑seq) analyses identified downstream targets involved in cell division, cell wall remodeling, and osmotic regulation that underpin the observed phenotypes. These findings reveal novel functions for PI paralogs in reproductive development and highlight mechanisms of transcription factor diversification in Oryza sativa.

The study compared arabinogalactan‑proteins (AGPs) from the hornwort Anthoceros agrestis and the moss Physcomitrium patens, revealing a galactan core lacking 1,6‑linked galactose and the presence of unusual 3‑O‑methyl‑rhamnose residues that are absent in angiosperms. Bioinformatic analyses across 14 bryophyte genomes showed limited GPI‑anchored AGP backbones in hornworts, while immunocytochemistry localized AGPs to plasma‑membrane, cell‑wall, vacuolar, and vesicle membranes, suggesting novel functions in early land plants.