The study benchmarked over 20 web‑based gRNA on‑target efficiency prediction tools against an experimental plant CRISPR editing dataset, finding several machine‑learning based tools whose scores strongly correlated with observed InDel frequencies. Additionally, the performance of popular platforms such as CRISPOR and CRISPR‑P was assessed, offering guidance for improved gRNA design in plant genome editing.

The study examined how polyploidy, hybridization, and incomplete lineage sorting affect phylogenetic reconstructions in the genus Packera, evaluating several published paralog‑processing pipelines. Results showed that the choice of orthology and paralog handling methods markedly altered tree topology, time‑calibrated phylogenies, biogeographic histories, and detection of ancient reticulation, underscoring the need for careful methodological selection alongside comprehensive taxon sampling.

The study identifies an autophagy pathway that degrades plasma membrane-derived clathrin-coated vesicles (CCVs) during hyperosmotic stress, helping maintain membrane tension as cell volume decreases. Using live imaging and correlative microscopy, the authors show that the TPLATE complex subunits AtEH1/Pan1 and AtEH2/Pan1 act as selective autophagy receptors by directly binding ATG8, thereby removing excess membrane under drought or salt conditions.

The study used Arabidopsis thaliana mutants with low (vtc2, vtc4) and high (vtc2/OE-VTC2) ascorbate levels to examine how ascorbate concentration affects gene expression and cellular homeostasis. Transcriptomic analysis revealed that altered ascorbate levels modulate defense and stress pathways, and that TAA1/TAR2‑mediated auxin biosynthesis is required for coping with elevated ascorbate in a light‑dependent manner.

Populus VariantDB v3.2 is a searchable database that catalogs sequence polymorphisms across heterozygous poplar genomes, enabling precise design of guide RNAs and primers for CRISPR genome editing in outcrossing tree species. The tool accepts short sequence inputs, supports multiple Populus genomes (including hybrids), and provides customizable search parameters to improve gRNA/primer design accuracy. Its platform‑agnostic and expandable architecture makes it broadly applicable to sequence‑sensitive functional genomics studies.