Plant regeneration methods for rapid generation of a large scale Ds transposant population in rice

Yuan Hu Xuan, Jin Huang, Gihwan Yi, Dong Soo Park, Soo Kwon Park, Moo Young Eun, Doh Won Yun, Gang Seob Lee, Tae Ho Kim, Chang Deok Han

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

1 Scopus citations

Abstract

To mutagenize rice genomes, a two-element system is utilized. This system comprises an immobile Ac element driven by the CaMV 35S promoter, and a gene trap Ds carrying a partial intron with alternative splice acceptors fused to the GUS coding region. Rapid, large-scale generation of a Ds transposant population was achieved using a plant regeneration procedure involving the tissue culture of seed-derived calli carrying Ac and Ds elements. During tissue cultures, Ds mobility accompanies changes in methylation patterns of a terminal region of Ds, where over 70 % of plants contained independent Ds insertions. In the transposon population, around 12 % of plants expressed GUS at the early seedling stage. A flanking-sequence-tag (FST) database has been established by cloning over 19,968 Ds insertion sites and the Ds map shows relatively uniform distribution across the rice chromosomes.

Original languageEnglish
Title of host publicationPlant Transposable Elements
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages101-116
Number of pages16
ISBN (Print)9781627035675
DOIs
StatePublished - 2013

Publication series

NameMethods in Molecular Biology
Volume1057
ISSN (Print)1064-3745

Keywords

  • Ac/Ds transposable elements
  • Gene trap
  • Mutagenesis
  • Plant regeneration
  • Rice
  • Tissue culture

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