An efficient field screening procedure for identifying transposants for constructing an Ac/Ds-based insertional-mutant library of rice

Wei Jiang Luan, Cheng Kun He, Guo Cheng Hu, Moul Dey, Ya Ping Fu, Hua Min Si, Li Zhu, Wen Zhen Liu, Faping Duan, Hong Zhang, Wen Ying Liu, Ren Ying Zhuo, Ajay Garg, Ray Wu, Zong Xiu Sun

Research output: Contribution to journalArticlepeer-review

7 Citations (Scopus)

Abstract

An efficient system was developed, and several variables tested, for generating a large-scale insertional-mutagenesis population of rice. The most important feature in this improved Ac/Ds tagging system is that one can conveniently carry out large-scale screening in the field and select transposants at the seedling stage. Rice was transformed with a plasmid that includes a Basta-resistance gene (bar). After the Ds element is excised during transposition, bar becomes adjacent to the ubiquitin promoter, and the rice plant becomes resistant to the herbicide Basta. In principle, one can plant up to one million plants in the field and select those plants that survive after spraying with Basta. To test the utility of this system, 4 Ds starter lines were crossed with 14 different Ac plants, and many transposants were successfully identified after planting 134 285 F2 plants in the field. Over 2 800 of these transposants were randomly chosen for PCR analysis, and the results fully confirmed the reliability of the field screening procedure.

Original languageEnglish
Pages (from-to)41-49
Number of pages9
JournalGenome
Volume51
Issue number1
DOIs
Publication statusPublished - Jan 2008
Externally publishedYes

Keywords

  • Ac/Ds transposon
  • Gene trap
  • Large-scale screening
  • Mutant library
  • Rice

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Genetics

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