Journal of Plant Biotechnology

The Korean Society of Plant Biotechnology (한국식물생명공학회)
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A Routine System for Generation of Fertile Transgenic Rice Plants Using Biolistic Method

  • Lee Soo-In (Division of Applied Life Science(BK 21 program), Gyeongsang National University) ;
  • Kim Cha-Young (Division of Applied Life Science(BK 21 program), Gyeongsang National University) ;
  • Lim Chae-Oh (Division of Applied Life Science(BK 21 program), Gyeongsang National University) ;
  • Choi Young-Ju (Department of Food and Nutrition, Silla University) ;
  • Kim Ho-Il (Genomics Division, National Institute of Agricultural Biotechnology) ;
  • Lee Sang-Yeol (Division of Applied Life Science(BK 21 program), Gyeongsang National University) ;
  • Lee Sung-Ho (Division of Applied Life Science(BK 21 program), Gyeongsang National University)
  • Published : 2003.09.01
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Abstract

A routine system based on particle bombardment of embryogenic callus for recovery of fertile transgenic rice (Oryza sativa L.) plants was developed. Embryogenic callus was established within 2-3 months from calli derived from mature seeds of Korean rice cultivar, Nagdongbyeo. The callus was bombarded with the plasmid pRQ6 containing the $\beta$-glucuronidase gene (gusA) and hygromycin phosphotransferase gene (hph, conferring resistance to hygromycin B), both driven by CaMV 35S promoter. Placement of cells on an osmoticum-containing medium (0.2 M sorbitol and 0.2 M mannitol) 4 hrs prior to and 16 hrs after bombardment resulted in a statistically significant increase with 3.2-fold in transient expression frequency gusA. In five independent experiments, the average frequency of transformation showing GUS activities was $8.86\%$. A large number of morphologically normal, fertile transgenic rice plants were obtained. Integration of foreign gene into the genome of $R_0$ transgenic plants was confirmed by Southern blot analysis. GUS and HPT were detected in $R_1$ progeny and Mendelian segregation of these genes was observed in $R_1$ progeny.

Keywords

References

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