Integrating Green Revolution Rht-1 Alleles in Diverse Wheat Landraces Using CRISPR/Cas9, Cytosine Base Editing and Prime Editing

Prime editing in genetically diverse bread wheat landraces enabled precise Rht-D1b-like G-to-T substitution, with a WATDE0394 plant showing 100% edited D-genome reads and 69 cm height.

• Why it matters: Landrace diversity is valuable for wheat breeding, but genome editing has been limited in diverse genetic backgrounds, restricting practical use of Rht-1 height variation.
• What they did: The team integrated CRISPR/Cas9 deletion, cytosine base editing, and prime editing in A.E. Watkins Collection landraces from AG1, AG3, AG4, AG6, and AG7, including a wheat-adapted TaCBE6b-intron system and an adapted ePPEplus prime editor.
• The result: Prime-editing activity varied by genotype (detected in WATDE0394, WATDE0549, WATDE0639, and WATDE0706), and edited Rht-D1 variants were linked to reduced height, with all T1 progeny of the 69 cm WATDE0394 plant homozygous for the prime edit by KASP.