Establishment of genetic transformation system via Agrobacterium in tall fescue cultivar

Abstract

Tall fescue (Festuca arundinacea Schreb.) is a cool-season turfgrass used on fairways in golf courses. The object of this study was to develop a more efficient, reliable, and repeatable approach in transforming the grass using Agrobacterium (EHA105), where β-glucuronidase gene (uidA) was used as a reporter and hygromycin phosphotransferase gene (hyg) as a selectable marker. An effective expression of transgene was observed in transforming 2-month-old calli derived from mature seeds (cv. Bingo) cultured on MS medium supplemented with 9 mg·L⁻¹ 2, 4-D. A two-step solid medium selection with increasing hygromycin concentration (from 30 to 50 mg·L⁻¹) was used to obtain resistant calli. Transgenic plants have been produced from many independent transformed calli. The presence of functional β-glucuronidase gene (uidA) was detected in hygromycin-resistant calli. Transgenic plants were regenerated and PCR and Southern blot confirmed transgene integration in the tall fescue genome.

Keywords

Tall Fescue / Agrobacterium tumefaciens / Transformation / β-glucuronidase gene (uidA)

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