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Plant Cell Advance Online Publication
Published on February 16, 2007; 10.1105/tpc.106.046938

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Received August 29, 2006
Returned for revision December 4, 2006
Accepted January 25, 2007

NUCLEOPORIN85 Is Required for Calcium Spiking, Fungal and Bacterial Symbioses, and Seed Production in Lotus japonicus

Katsuharu Saito 1, Makoto Yoshikawa 2, Koji Yano 2, Hiroki Miwa 3, Hisaki Uchida 4, Erika Asamizu 5, Shusei Sato 5, Satoshi Tabata 5, Haruko Imaizumi-Anraku 6, Yosuke Umehara 6, Hiroshi Kouchi 6, Yoshikatsu Murooka 2, Krzysztof Szczyglowski 7, J. Allan Downie 3, Martin Parniske 8, Makoto Hayashi 9, and Masayoshi Kawaguchi 10*

1 Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan
2 Department of Biotechnology, Graduate School of Engineering, Osaka University, Osaka 565-0871, Japan
3 Department of Molecular Microbiology, John Innes Centre, Norwich NR4 7UH, United Kingdom
4 Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan
5 Kazusa DNA Research Institute, Chiba 292-0818, Japan
6 Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan; Plant Physiology Department, National Institute of Agrobiological Sciences, Ibaraki 305-8602, Japan
7 Agriculture and Agri-Food Canada, Southern Crop Protection and Food Research Centre, Ontario, N5V 4T3, Canada
8 Department of Biology, Genetics, University of Munich, D-80638 Munich, Germany
9 Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan; Department of Biotechnology, Graduate School of Engineering, Osaka University, Osaka 565-0871, Japan
10 Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama 332-0012, Japan; Department of Biological Sciences, Graduate School of Science, University of Tokyo, Tokyo 113-0033, Japan

* To whom correspondence should be addressed. E-mail: masayosi{at}biol.s.u-tokyo.ac.jp.

In Lotus japonicus, seven genetic loci have been identified thus far as components of a common symbiosis (Sym) pathway shared by rhizobia and arbuscular mycorrhizal fungi. We characterized the nup85 mutants (nup85-1, -2, and -3) required for both symbioses and cloned the corresponding gene. When inoculated with Glomus intraradices, the hyphae managed to enter between epidermal cells, but they were unable to penetrate the cortical cell layer. The nup85-2 mutation conferred a weak and temperature-sensitive symbiotic phenotype, which resulted in low arbuscule formation at 22°C but allowed significantly higher arbuscule formation in plant cortical cells at 18°C. On the other hand, the nup85 mutants either did not form nodules or formed few nodules. When treated with Nod factor of Mesorhizobium loti, nup85 roots showed a high degree of root hair branching but failed to induce calcium spiking. In seedlings grown under uninoculated conditions supplied with nitrate, nup85 did not arrest plant growth but significantly reduced seed production. NUP85 encodes a putative nucleoporin with extensive similarity to vertebrate NUP85. Together with symbiotic nucleoporin NUP133, L. japonicus NUP85 might be part of a specific nuclear pore subcomplex that is crucial for fungal and rhizobial colonization and seed production.




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