Precision recreation and multiplex stacking of disease-resistance alleles in tomato using optimized prime editing.

Optimized prime editing in tomato (Solanum lycopersicum) precisely recreated disease-resistance alleles, including deletion of two leucine residues in SlBs5/SlBs5L, yielding enhanced Xanthomonas resistance.

• Why it matters: Efficient, multiplexable prime editing in dicot crops is still limited, slowing the ability to recreate and stack beneficial resistance alleles for breeding.
• What they did: The study optimized three tomato prime editors (TomPE2, TomPE6c, TomPE7) to edit SlBs5/SlBs5L and SlDMR6, then compared editor architectures and culture temperatures and used transcriptome profiling to define defense states.
• The result: TomPE6c produced the highest proportion of precise edits, multiplex editing enabled combining resistance and regulatory variants in one generation, and engineered alleles were stably inherited with resistance maintained under elevated temperature conditions.

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