Anirudh N, Haritha P K, Sreedhar R V
Plant Cell Biotechnology Department, Council of Scientific and Industrial Research-Central Food Technological Research Institute, Mysuru, Karnataka, 570020, India.
Academy of Scientific and Innovative Research (AcSIR), Ghaziabad, 201002, India.
Transgenic Res. 2025 Jun 18;34(1):31. doi: 10.1007/s11248-025-00450-z.
Chia (Salvia hispanica L.), a nutritionally 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 protocol optimized for high-efficiency transformation of chia seedlings. Key parameters including bacterial cell density, acetosyringone concentration, sonication duration, vacuum infiltration, and infection time were optimized. Results demonstrated that an optical density at 600 nm (OD) of 0.5, acetosyringone concentration of 100 µM, 20 min of sonication, 10 min of vacuum infiltration, and 60 min of infection 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 transformation protocol, paving the way for future genetic engineering efforts aimed at enhancing the nutritional and agronomic traits of chia.
奇亚（Salvia hispanica L.）是一种具有重要营养价值的作物，是关键的ω-3脂肪酸α-亚麻酸的最丰富来源。尽管奇亚具有营养价值，但目前尚无稳定的遗传转化方法。本研究提出了一种经超声辅助的农杆菌介导的转化方案，该方案针对奇亚幼苗的高效转化进行了优化。对包括细菌细胞密度、乙酰丁香酮浓度、超声处理时间、真空渗透和侵染时间等关键参数进行了优化。结果表明，600nm处的光密度（OD）为0.5、乙酰丁香酮浓度为100µM、超声处理20分钟、真空渗透10分钟和侵染60分钟可显著提高转化效率和GUS表达。通过聚合酶链反应和对转化植株的β-葡萄糖醛酸酶（GUS）检测对该优化方案进行了验证。我们的研究结果建立了一种强大且可靠的转化方案，为未来旨在改善奇亚营养和农艺性状的基因工程努力铺平了道路。
