title: Rapid, large-scale generation of Ds transposant lines and analysis of the Ds insertion sites in rice.
abstract: Rapid, large-scale generation of a Ds transposant population was achieved using a regeneration procedure involving tissue culture of seed-derived calli carrying Ac and inactive Ds elements. In the F(2) progeny from genetic crosses between the same Ds and Ac starter lines, most of the crosses produced an independent germinal transposition frequency of 10-20%. Also, many Ds elements underwent immobilization even though Ac was expressed. By comparison, in a callus-derived regenerated population, over 70% of plants carried independent Ds insertions, indicating transposition early in callus formation. In the remaining population, the majority of plants carried only Ac. Most of the new Ds insertions were stably transmitted to a subsequent generation. An exceptionally high proportion of independent transposants in the regenerated population means that selection markers for transposed Ds and continual monitoring of Ac/Ds activities may not necessarily be required. By analyzing 1297 Ds-flanking DNA sequences, a genetic map of 1072 Ds insertion sites was developed. The map showed that Ds elements were transposed onto all of the rice chromosomes, with preference not only near donor sites (36%) but also on certain physically unlinked arms. Populations from both genetic crossing and tissue culture showed the same distribution patterns of Ds insertion sites. The information of these mapped Ds insertion sites was deposited in GenBank. Among them, 55% of Ds elements were on predicted open-reading frame (ORF) regions. Thus, we propose an optimal strategy for the rapid generation of a large population of Ds transposants in rice.
authors.affiliation: Division of Applied Life Science, BK21 Program, Plant Molecular Biology & Biotechnology Research Center (PMBBRC), Gyeongsang National University, Jinju 660-701, Korea.
authors.forename: Chul Min
authors.initials: CM
authors.lastname: Kim
authors.forename: Hai Long
authors.initials: HL
authors.lastname: Piao
authors.forename: Soon Ju
authors.initials: SJ
authors.lastname: Park
authors.forename: Nam Soo
authors.initials: NS
authors.lastname: Chon
authors.forename: Byoung Il
authors.initials: BI
authors.lastname: Je
authors.forename: Bingyao
authors.initials: B
authors.lastname: Sun
authors.forename: Sung Han
authors.initials: SH
authors.lastname: Park
authors.forename: Jin Young
authors.initials: JY
authors.lastname: Park
authors.forename: Eun Jin
authors.initials: EJ
authors.lastname: Lee
authors.forename: Min Jung
authors.initials: MJ
authors.lastname: Kim
authors.forename: Woo Sik
authors.initials: WS
authors.lastname: Chung
authors.forename: Kon Ho
authors.initials: KH
authors.lastname: Lee
authors.forename: Young Suk
authors.initials: YS
authors.lastname: Lee
authors.forename: Jeung Joo
authors.initials: JJ
authors.lastname: Lee
authors.forename: Yong Jae
authors.initials: YJ
authors.lastname: Won
authors.forename: GiHwan
authors.initials: G
authors.lastname: Yi
authors.forename: Min Hee
authors.initials: MH
authors.lastname: Nam
authors.forename: Young Soon
authors.initials: YS
authors.lastname: Cha
authors.forename: Doh Won
authors.initials: DW
authors.lastname: Yun
authors.forename: Moo Young
authors.initials: MY
authors.lastname: Eun
authors.forename: Chang-deok
authors.initials: CD
authors.lastname: Han
mesh_terms: D002874:Chromosome Mapping; D003433:Crosses, Genetic; D046508:Culture Techniques; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D018745:Genome, Plant; D008957:Models, Genetic; D016254:Mutagenesis, Insertional; D012275:Oryza / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012038:Regeneration; D012639:Seeds / Q000235:genetics / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D018744:DNA, Plant; C036483:T-DNA