Activation tagging approach in a model legume, Lotus japonicus

Ryujiro Imaizumi, Shusei Sato, Nanako Kameya, Ikuo Nakamura, Yasukazu Nakamura, Satoshi Tabata, Shin Ichi Ayabe, Toshio Aoki

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

We constructed T-DNA insertional lines of a model legume, Lotus japonicus, using a multifunctional vector for gene and exon activation tagging. The vector had the CaMV 35S promoter together with two additional enhancer elements, the start codon, and splice donor and acceptor sites facing the left border, in anticipation of the activation of T-DNA flanking genes and forced expression of flanking exons. The improved transformation technique yielded more than 3,500 lines, including 45 dominant mutant candidates with abnormal phenotypes with respect to aerial parts, nodules, and roots. Among the 44 selected lines, one copy of T-DNA was inserted into the genome of 37 lines (84%). The T-DNA flanking regions of seven lines were isolated by thermal asymmetric interlaced (TAIL)-PCR or reverse transcription (RT)-PCR, and the corresponding genomic clones were analyzed. The transcripts of four genes adjacent to T-DNA out of 11 genes tested were increased in the T1 generation, demonstrating that gene and exon activation effects by the newly developed tagging vector are heritable. The T-DNA insertional population of L. japonicus will provide legume-specific dominant mutants.

Original languageEnglish
Pages (from-to)391-399
Number of pages9
JournalJournal of Plant Research
Volume118
Issue number6
DOIs
Publication statusPublished - 2005 Dec 1
Externally publishedYes

Keywords

  • Activation tagging
  • Legume
  • Lotus japonicus
  • Thermal asymmetric interlaced (TAIL)-PCR

ASJC Scopus subject areas

  • Plant Science

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  • Cite this

    Imaizumi, R., Sato, S., Kameya, N., Nakamura, I., Nakamura, Y., Tabata, S., Ayabe, S. I., & Aoki, T. (2005). Activation tagging approach in a model legume, Lotus japonicus. Journal of Plant Research, 118(6), 391-399. https://doi.org/10.1007/s10265-005-0231-5