The authors introduce the ENABLE(R) Gene Editing in planta toolkit, a streamlined two‑step cloning system for creating CRISPR/Cas9 knockout vectors suitable for transient or stable transformation. Validation was performed in Oryza sativa protoplasts and Arabidopsis thaliana plants, and the toolkit includes low‑cost protocols aimed at facilitating adoption in the Global South.

The study generated a phenotypic dataset for 550 Lactuca accessions, including 20 wild relatives, and applied an iterative two‑step GWAS using a jointly processed SNP set for cultivated lettuce (L. sativa) and its wild progenitor (L. serriola) to dissect trait loci. Known and novel QTLs for anthocyanin accumulation, leaf morphology, and pathogen resistance were identified, with several L. serriola‑specific QTLs revealing unique genetic architectures, underscoring the breeding value of wild lettuce species.