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KMID : 0613820130230111317
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Journal of Life Science
2013 Volume.23 No. 11 p.1317 ~ p.1324
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Development of Marker-free Transgenic Rice for Increasing Bread-making Quality using Wheat High Molecular Weight Glutenin Subunits (HMW-GS) Gene
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Park Soo-Kwon
Shin Dong-Jin
Hwang Woon-Ha
Oh Se-Yun
Cho Jun-Hyun
Han Sang-Ik
Nam Min-Hee
Park Dong-Soo
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Abstract
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High-molecular weight glutenin subunits (HMW-GS) have been shown to play a crucial role in determining the processing properties of the wheat grain. We have produced marker-free transgenic rice plants containing a wheat Glu-1Bx7 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using the Agrobacterium-mediated co-transformation method. The Glu-1Bx7-own promoter was inserted into a binary vector for seed-specific expression of the Glu-1Bx7 gene. Two expression cassettes comprised of separate DNA fragments containing only Glu-1Bx7 and hygromycin phosphotransferase II (HPTII) resistance genes were introduced separately to the Agrobacterium tumefaciens EHA105 strain for co-infection. Each EHA105 strain harboring Glu-1Bx7 or HPTII was infected to rice calli at a 3:1 ratio of Glu-1Bx7 and HPTII, respectively. Then, among 216 hygromycin-resistant T_0 plants, we obtained 24 transgenic lines with both Glu-1Bx7 and HPTII genes inserted into the rice genome. We reconfirmed integration of the Glu-1Bx7 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the wheat Glu-1Bx7 were stably expressed in the rice T₁ seeds. Finally, the marker-free plants harboring only the Glu-1Bx7 gene were successfully screened at the T₁ generation.
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KEYWORD
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Co-transformation, high-molecular-weight glutenin subunit (HMW-GS) protein, marker-free transgenic rice, wheat
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