Bioactive beads-mediated transformation of rice with large DNA fragments containing Aegilops tauschii genes.
Transformation with large DNA molecules enables multiple genes to be introduced into plants simultaneously to produce transgenic plants with complex phenotypes. In this study, a large DNA fragment (ca. 100 kb) containing a set of Aegilops tauschii hardness genes was introduced into rice plants using a novel transformation method, called bioactive beads-mediated transformation. Nine transgenic rice plants were obtained and the presence of transgenes in the rice genome was confirmed by PCR and FISH analyses. The results suggested that multiple transgenes were successfully integrated in all transgenic plants. The expression of one of the transgenes, puroindoline b, was confirmed at the mRNA and protein levels in the T(2) generation. Our study clearly demonstrates that the bioactive bead method is capable of producing transgenic rice plants carrying large DNA fragments. This method will facilitate the production of useful transgenic plants by introducing multiple genes simultaneously.
Wada
Naoki
N
Department of Biotechnology, Graduate School of Engineering, Osaka University, Suita, Osaka, Japan.
Kajiyama
Shin'ichiro
S
Akiyama
Yukio
Y
Kawakami
Shigeki
S
No
Daisuke
D
Uchiyama
Susumu
S
Otani
Motoyasu
M
Shimada
Takiko
T
Nose
Naoko
N
Suzuki
Go
G
Mukai
Yasuhiko
Y
Fukui
Kiichi
K
D022202:Chromosomes, Artificial, Bacterial; D002934:Cinnamates / Q000494:pharmacology; D018744:DNA, Plant / Q000235:genetics; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017343:Genes, Plant; D005822:Genetic Vectors; D006921:Hygromycin B / Q000031:analogs & derivatives / Q000494:pharmacology; D008863:Microspheres; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D006109:Poaceae / Q000235:genetics; D014170:Transformation, Genetic; D019076:Transgenes
D002934:Cinnamates; D018744:DNA, Plant; D006921:Hygromycin B; C026273:hygromycin A