Molecular Genetics and Genomics

, Volume 279, Issue 3, pp 213–223

MNU-induced mutant pools and high performance TILLING enable finding of any gene mutation in rice

  • Tadzunu Suzuki
  • Mitsugu Eiguchi
  • Toshihiro Kumamaru
  • Hikaru Satoh
  • Hiroaki Matsusaka
  • Kazuki Moriguchi
  • Yasuo Nagato
  • Nori Kurata
Original Paper

DOI: 10.1007/s00438-007-0293-2

Cite this article as:
Suzuki, T., Eiguchi, M., Kumamaru, T. et al. Mol Genet Genomics (2008) 279: 213. doi:10.1007/s00438-007-0293-2

Abstract

Mutant populations are indispensable genetic resources for functional genomics in all organisms. However, suitable rice mutant populations, induced either by chemicals or irradiation still have been rarely developed to date. To produce mutant pools and to launch a search system for rice gene mutations, we developed mutant populations of Oryza sativajaponica cv. Taichung 65, by treating single zygotic cells with N-methyl-N-nitrosourea (MNU). Mutagenesis in single zygotes can create mutations at a high frequency and rarely forms chimeric plants. A modified TILLING system using non-labeled primers and fast capillary gel electrophoresis was applied for high-throughput detection of single nucleotide substitution mutations. The mutation rate of an M2 mutant population was calculated as 7.4 × 10−6 per nucleotide representing one mutation in every 135 kb genome sequence. One can expect 7.4 single nucleotide substitution mutations in every 1 kb of gene region when using 1,000 M2 mutant lines. The mutations were very evenly distributed over the regions examined. These results indicate that our rice mutant population generated by MNU-mutagenesis could be a promising resource for identifying mutations in any gene of rice. The modified TILLING method also proved very efficient and convenient in screening the mutant population.

Keywords

MNU-mutagenesis Rice Zygote cell SNP detection High throughput TILLING 

Supplementary material

438_2007_293_MOESM1_ESM.pdf (1.8 mb)
(PDF 1.83 Mb)

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Tadzunu Suzuki
    • 1
  • Mitsugu Eiguchi
    • 1
  • Toshihiro Kumamaru
    • 2
  • Hikaru Satoh
    • 2
  • Hiroaki Matsusaka
    • 2
  • Kazuki Moriguchi
    • 1
    • 3
  • Yasuo Nagato
    • 4
  • Nori Kurata
    • 1
    • 5
  1. 1.Genetic Strains Research CenterNational Institute of GeneticsShizuokaJapan
  2. 2.Faculty of AgricultureKyushu UniversityFukuokaJapan
  3. 3.Graduate School of ScienceHiroshima UniversityHigashi-HiroshimaJapan
  4. 4.Graduate School of Agriculture and Life SciencesUniversity of TokyoTokyoJapan
  5. 5.School of Life ScienceGraduate University of SOKENDAIShizuokaJapan

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