A rice retrotransposon, Tos17, as a tool for gene tagging
2003
Murata, K. | Miyao, A. | Tanaka, K. | Yamazaki, M. | Takeda, S. | Abe, K. | Onosato, K. | Miyazaki, A. | Yamashita, Y. | Sasaki, T. | Hirochika, H..
A rice retrotransposon, Tos17, possesses unique features: transposition induced by tissue culture, a low copy number (1-4), high frequency of transposition, resulting in 5-30 transposed copies, and random transposition throughout the genome. In regenerated plants, transposed Tos17 copies are inactivated. Thus, Tos17 could be used for insertional mutagenesis as a tool for functional analysis of rice genes. We have constructed a rice mutant panel consisting of more than 30,000 regenerated knockout lines. Phenotypes of 2,300 lines were investigated in the field using R2 segregating populations. Many kinds of mutant phenotypes were observed at various stages, about 30% of the lines showed mutant phenotypes on plant morphology and degree of fertility. Southern blot analysis was performed to identify the specific Tos17 copy cosegregating with the mutant phenotype. The results strongly suggested that some mutations, including narrow leaf, brittle culm, viviparous, stripe, dwarf, albino, low fertility, and sterility, were caused by a Tos17 insertion. Genomic sequences corresponding to their mutant loci were amplified by TAIL-polymerase chain reaction (PCR) or suppression-PCR, cloned, and partially determined. A similarity search of determined sequence data suggested that some of them have high similarity to known genes such as genes for cellulose synthases and transcription factor. The others were new genes such as the gene regulating leaf width in the narrow-leaf mutant.
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