Perilla transformation using selection markers containing antibiotics and basta
2008
Kim, K.H. (National Academy of Agricultural Science, RDA, Suwon, Republic of Korea), E-mail: kimhwan@rda.go.kr | Lee, J.E. (National Academy of Agricultural Science, RDA, Suwon, Republic of Korea) | Ha, S.H. (National Academy of Agricultural Science, RDA, Suwon, Republic of Korea) | Hahn, B.S. (National Academy of Agricultural Science, RDA, Suwon, Republic of Korea) | Park, J.S. (National Academy of Agricultural Science, RDA, Suwon, Republic of Korea) | Lee, M.H. (National Institute of Crop Science, RDA, Milyang, Republic of Korea) | Jung, C.S. (National Institute of Crop Science, RDA, Milyang, Republic of Korea) | Kim, Y.H. (National Academy of Agricultural Science, RDA, Suwon, Republic of Korea)
A modified method of Agrobacterium-mediated perilla transformation was developed using two selection markers of an antibiotics (either hpt or nptll) and an herbicidal (bar) gene. Perilla hypocotyl explants were cocultured with Agrobacterium tumefaciens EHA 105 strain harboring plasmid vector (either pMOG6-Bar or pCK-Bar) for three days, respectively. Primary shoots were selected with antibiotics of hygromycin (15 mg/L) or kanamycin (125 mg/L) and regenerated shoots were further selected with herbicide phosphinothricin (ppt, 1.2 mg/L) to obtain authentic transformants. Roots were induced for the regenerated shoots on the MS medium without hormone and 80 putative transgenic plants were obtained. Transgene integration into perilla genome was confirmed by Southern blot and their expression was analyzed by Northern blot. T1 perilla seeds drived from To plants were tested 0.3% basta spray for identification of stable gene delivery to next generation.
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