Regeneration of transgenic Cryptomeria japonica D. Don after Agrobacterium tumefaciens-mediated transformation of embryogenic tissue.
A genetic transformation procedure for Cryptomeria japonica was developed after co-cultivation of embryogenic tissues with the disarmed Agrobacterium tumefaciens strain C58/pMP90, which harbours the visual reporter gene sgfp and two selectable marker genes, hpt and nptII. We were able to generate eight and three independent transgenic lines per gram of embryogenic tissue after selection on hygromycin and kanamycin medium, respectively. Transgenic plants were regenerated through somatic embryogenesis in 4 lines out of these 11 lines. Green fluorescent protein fluorescence was observed under fluorescent microscopy. Integration of the genes into the genome was confirmed by polymerase chain reaction analysis of embryogenic tissues and Southern blot analysis of regenerated plantlets.
Forest Bio-Research Center, Forest and Forestry Product Research Institute (FFPRI), 3809-1 Ishi, Juo, Hitachi, Ibaraki, 319-1301, Japan. toru.t@affrc.go.jp
Toru
T
Taniguchi
Yasunori
Y
Ohmiya
Manabu
M
Kurita
Miyoko
M
Tsubomura
Teiji
T
Kondo
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D002934:Cinnamates / Q000494:pharmacology; D032485:Cryptomeria / Q000187:drug effects / Q000196:embryology / Q000235:genetics*; D018744:DNA, Plant / Q000235:genetics; D047108:Embryonic Development; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D007612:Kanamycin / Q000494:pharmacology; D030821:Plants, Genetically Modified / Q000187:drug effects / Q000196:embryology / Q000235:genetics*; D012038:Regeneration; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic
D002934:Cinnamates; D018744:DNA, Plant; D049452:Green Fluorescent Proteins; D006921:Hygromycin B; C026273:hygromycin A; D007612:Kanamycin