Agrobacterium-mediated genetic transformation of Perilla frutescens.
A reproducible plant regeneration and an Agrobacterium tumefaciens-mediated genetic transformation protocol were developed for Perilla frutescens (perilla). The largest number of adventitious shoots were induced directly without an intervening callus phase from hypocotyl explants on MS medium supplemented with 3.0 mg/l 6-benzylaminopurine (BA). The effects of preculture and extent of cocultivation were examined by assaying beta-glucuronidase (GUS) activity in explants infected with A. tumefaciens strain EHA105 harboring the plasmid pIG121-Hm. The highest number of GUS-positive explants were obtained from hypocotyl explants cocultured for 3 days with Agrobacterium without precultivation. Transgenic perilla plants were regenerated and selected on MS basal medium supplemented with 3.0 mg/l BA, 125 mg/l kanamycin, and 500 mg/l carbenicillin. The transformants were confirmed by PCR of the neomycin phosphotransferase II gene and genomic Southern hybridization analysis of the hygromycin phosphotransferase gene. The frequency of transformation from hypocotyls was about 1.4%, and the transformants showed normal growth and sexual compatibility by producing progenies.
Kim
Kyung-Hwan
KH
National Institute of Agricultural Biotechnology, Rural Development Administration, Suwon, 441-707, South Korea.
Lee
Yeon-Hee
YH
Kim
Donghern
D
Park
Yong-Hwan
YH
Lee
Jai-Youl
JY
Hwang
Young-Soo
YS
Kim
Yong-Hwan
YH
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D031344:Perilla frutescens / Q000235:genetics* / Q000254:growth & development; D010641:Phenotype; D030821:Plants, Genetically Modified / Q000254:growth & development; D036226:Seedlings / Q000235:genetics / Q000254:growth & development; D014170:Transformation, Genetic