PRODUCTION AND ANALYSIS OF RICE RETROTRANSPOSON Tos17 INSERTION LINES

¹ National Institute of Agrobiological Resources, Tsukuba, Ibaraki 305-8602, Japan

² Institute of Society for Techno-innovation of Agriculture, Forestry and Fisheries, Tsukuba, Ibaraki 305-0854, Japan

The rice retrotransposon Tos17 is specifically activated by tissue culture and activated copies are inserted anywhere in the rice genome. Since the activated Tos17 copies become inactive in regenerated plants and inactivated Tos17 copies are fixed at inserted positions, Tos17 will be used for insertional mutagenesis as a tool for functional analysis of rice genes. We have made 8885 regenerated Nipponbare lines and stocked 431 Akitakomachi and 838 Hitomebore lines. Each line of Nipponbare regenerated plants has average 5.5 copies of newly transposed Tos17. Insertion sites have been isolated by TAIL-PCR and suppression PCR method. Nucleotide sequences were determined by direct sequencing of PCR products and analyzed by similarity search against non-redundant DNA database. Insertion mutants of different classes of genes have been identified and mutant phenotypes of some of them analyzed for functional analysis of genes. For example, homozyous mutants of H0054, in which aurora-related protein kinase gene is disrupted, showed a sterile phenotype and reduced growth.