Transgene‐free genome editing in poplar

• DOI: 10.1111/nph.20415
• Publication Date: 2025-01-22T17:54:31Z
• Authors: Lennart Hoengenaert; Chantal Anders; Jan Van Doorsselaere; Ruben Vanholme; Wout Boerjan
• Keywords: Marker gene

Summary Precise gene‐editing methods are valuable tools to enhance genetic traits. Gene editing is commonly achieved via stable integration of a cassette in the plant's genome. However, this technique unfavorable for field applications, especially vegetatively propagated plants, such as many commercial tree species, where cannot be segregated away without breaking constitution elite variety. Here, we describe an efficient method generating gene‐edited Populus tremula × P. alba (poplar) trees incorporating foreign DNA into its Using Agrobacterium tumefaciens , expressed base‐editing construct targeting CCoAOMT1 along with ALS genes positive selection on chlorsulfuron‐containing medium. About 50% regenerated shoots were derived from transient transformation and free T‐DNA. Overall, 7% chlorsulfuron‐resistant T‐DNA free, edited gene nonchimeric. Long‐read whole‐genome sequencing confirmed absence any tested lines. Additionally, evaluated CodA negative marker eliminate lines that stably incorporated their Although latter not essential selecting transgene‐free, shoots, it may prove other genotypes or varieties.