Agrobacterium‑mediated genetic transformation of chia (Salvia hispanica L.), a rich source of omega‑3 fatty acid

Chia (Salvia hispanica L.), a nutrition-
ally valuable crop, is the richest source of α-linolenic
acid, a key omega-3 fatty acid. Despite its nutritional
benefits, a stable genetic transformation method for
chia is not available. This study presents a sonication-
assisted Agrobacterium-mediated transformation pro-
tocol optimized for high-efficiency transformation of
chia seedlings. Key parameters including bacterial
cell density, acetosyringone concentration, sonica-
tion duration, vacuum infiltration, and infection time
were optimized. Results demonstrated that an opti-
cal density at 600 nm (OD600) of 0.5, acetosyrin-
gone concentration of 100 μM, 20 min of sonication,
10 min of vacuum infiltration, and 60 min of infec-
tion significantly enhanced transformation efficiency
and GUS expression. This optimized protocol was
validated through Polymerase Chain Reaction and
β-glucuronidase (GUS) assay in transformed plants.
Our findings establish a robust and reliable transfor-
mation protocol, paving the way for future genetic engineering efforts aimed at enhancing the nutritional
and agronomic traits of chia.