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DNA Research Advance Access originally published online on January 22, 2007
DNA Research 2006 13(6):255-265; doi:10.1093/dnares/dsl017
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© The Author 2006. Kazusa DNA Research Institute.
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Positional Cloning Identifies Lotus japonicus NSP2, A Putative Transcription Factor of the GRAS Family, Required for NIN and ENOD40 Gene Expression in Nodule Initiation

Yasuhiro Murakami1,3,4, Hiroki Miwa2 {dagger}, Haruko Imaizumi-Anraku1,4, Hiroshi Kouchi1, J. Allan Downie2, Masayoshi Kawaguchi3,* and Shinji Kawasaki1,4

1 National Institute of Agrobiological Sciences, 2-1-2 Kannondai Tsukuba, Ibaraki 305-8602, Japan
2 John Innes Centre Colney Lane, Norwich NR4 7UH, UK
3 Department of Biological Sciences, Graduate School of Science, The University of Tokyo 7-3-1 Hongo, Bunkyo, Tokyo 113-0033, Japan
4 Bio-oriented Technology Research Advancement Institution (BRAIN), Tokyo Office 3-18-19 Toranomon, Minato-ku, Tokyo 105-0001, Japan

Rhizobia-secreted Nod-factors (NFs) are required for nodulation. In the early developmental process of nodulation, a large number of changes occur in gene expression. Lotus japonicus nsp2 mutants isolated from Gifu B-129 ecotype have defects in nodule initiation and display non-nodulating phenotype. Here, we describe positional cloning of LjNSP2 as a component of the nodulation-specific signaling pathway. LjNSP2 was mapped near the translocation site of chromosome 1 where the recombination is severely suppressed. To circumvent this problem, we introduced Lotus burttii as an alternative crossing partner in place of L. japonicus Miyakojima. The development of the high-resolution map using a total of 11 481 F2 plants, in combination with newly developed DNA markers and construction of BAC library, enabled us to identify the gene responsible for mutant phenotype. LjNSP2 encodes a putative transcription factor of the GRAS family that constitutes a subfamily with Medicago truncatula NSP2. LjNSP2 was expressed in roots and early nodules, but strongly suppressed in matured nodules. The expression analysis of NIN and LjENOD40-1 genes in Ljnsp2 mutants indicates that LjNSP2 functions upstream of these genes. These results suggest that LjNSP2 acts as a transcription factor to directly or indirectly switch on the NF-induced genes required for nodule initiation.

Key words: positional cloning; Lotus japonicus; transcription factor; nodule initiation

*To whom correspondence should be addressed. Tel./Fax: +81-3-5841-4458, E-mail: masayosi{at}biol.s.u-tokyo.ac.jp

{dagger}Present address: Biotechnology Research Center, The University of Tokyo, 1-1-1 Yayoi, Bunkyo, Tokyo 113-8657, Japan

Communicated by Satoshi Tabata


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[Abstract] [Full Text] [PDF]


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