Agrobacterium-mediated transformation of creeping bentgrass using GFP as areporter gene

Creeping bentgrass (Agrostis palustris Huds.) is a cool season grass widely used on putting greens in golf courses. Transformation of creeping bentgra ss has been conducted using microprojectile bombardment and protoplast elec troporation. The objective of our study is to develop an alternative and mo re efficient approach in transforming the grass using Agrobacterium (strain EHA 101). This technique was effective in transforming 40-day old calli de rived from mature seeds cultured on MS medium supplemented with 2,4-D, kine tin, and sucrose. Dozens of transgenic plants have been produced from two i ndependent transformed calli. Presence of functional green fluorescence pro tein (GFP) was detected in leaves, stems, and roots of transgenic seedlings . Four putative transgenic plants and two control plants were randomly chos en and analyzed by Southern blot analysis. Bands corresponding to the GFP g ene were clearly shown in transgenic plants. These results indicated that A grobacterium transformation can successfully be applied to creeping bentgra ss.