Field experiments combined with RNA sequencing revealed that wheat ploidy influences heat stress resilience, with tetraploid T. turgidum showing the smallest yield loss and hexaploid T. aestivum mounting the largest transcriptional response. Ploidy-dependent differences were observed in differential gene expression, alternative splicing—including hexaploid-specific exon skipping of NF‑YB—and co‑expression networks linked to grain traits, highlighting candidate pathways for breeding heat‑tolerant wheat.

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.