Highly Efficient <i>Agrobacterium tumefaciens</i> Mediated Transformation of Oil Palm Using an <i>EPSPS</i>-Glyphosate Selection System

Abstract

Oil palm (Elaeis guineensis Jacq.) is the most efficient oil-producing crop globally, yet progress in its research has been hampered by the lack of effective genetic transformation systems. The EPSPS gene, encoding 5-enolpyruvylshikimate-3-phosphate synthase, has been used as a transgenic selection marker in various crops, including rice and soybean. This study evaluated the EPSPS/glyphosate selection system for oil palm transformation. We constructed a binary expression vector, pCGlyDESCLI-C, containing the TIPS-EiEPSPS selection marker from goosegrass and the mScarlet-I red fluorescent reporter gene. This vector was introduced into oil palm embryonic callus (EC) via Agrobacterium-mediated transformation. After optimizing the transformation steps, positive calli were obtained, and integration of the foreign gene into the oil palm genome was confirmed through molecular analysis. Notably, the selection efficiency of the EPSPS/glyphosate selection system exceeded that of the traditional hpt/hygromycin selection system, demonstrating its advantages. Our findings support the effectiveness of the TIPS-EiEPSPS/glyphosate selection system for oil palm genetic transformation, marking its first application in this species and offering a valuable tool for advancing research on this economically significant crop.

Keywords

• oil palm
• genetic transformation
• embryonic callus
• <i>EPSPS</i>
• glyphosate
• <i>Agrobacterium</i>-mediated