A fluorescent antibiotic resistance marker for rapid production of transgenic rice plants.
Blasticidin S (BS) is an aminoacylnucleoside antibiotic used for the control of rice blast disease. To establish a new cereal transformation system, we constructed a visual marker gene designated gfbsd, encoding an enhanced green fluorescent protein (EGFP) fused to the N-terminus of BS deaminase (BSD). It was cloned into a monocot expression vector and introduced into rice (Oryza sativa L. cv. Nipponbare) calluses by microprojectile bombardment. Three to five weeks after the bombardment, multicellular clusters emitting bright-green EGFP fluorescence were obtained with 10 microg/ml BS, which is not sufficient to completely inhibit the growth of non-transformed tissues. Fluorescent sectors (approximately 2mm in diameter) excised from the calluses regenerated into transgenic plantlets (approximately 10 cm in height) as early as 51 (average 77+/-11) days after the bombardment. The visual antibiotic selection was more efficient and required less time than the bialaphos selection with bar. In addition, the small size (1.1 kb) of gfbsd is preferable for construction of transformation vectors. This new marker gene will make a significant contribution in molecular genetic studies of rice plants.
Ochiai-Fukuda
Tetsuko
T
Laboratory for Remediation Research, Plant Science Center (PSC1) and Discovery Research Institute (DRI), RIKEN, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan.
Takahashi-Ando
Naoko
N
Ohsato
Shuichi
S
Igawa
Tomoko
T
Kadokura
Kaori
K
Hamamoto
Hiroshi
H
Nakasako
Masayoshi
M
Kudo
Toshiaki
T
Shibata
Takehiko
T
Yamaguchi
Isamu
I
Kimura
Makoto
M
D001483:Base Sequence; D015415:Biomarkers / Q000378:metabolism; D004351:Drug Resistance; D005456:Fluorescent Dyes / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008957:Models, Genetic; D008969:Molecular Sequence Data; D012275:Oryza / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism*; D014170:Transformation, Genetic
D015415:Biomarkers; D005456:Fluorescent Dyes; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins