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PNAS 103 (2): 359-364

Copyright © 2006 by the National Academy of Sciences.

From The Cover


A nucleoporin is required for induction of Ca2+ spiking in legume nodule development and essential for rhizobial and fungal symbiosis

Norihito Kanamori *, {dagger}, Lene Heegaard Madsen *, Simona Radutoiu *, Mirela Frantescu *, Esben M. H. Quistgaard *, Hiroki Miwa {ddagger}, J. Allan Downie {ddagger}, Euan K. James §, Hubert H. Felle ¶, Line Lindegaard Haaning *, Torben Heick Jensen *, Shusei Sato ||, Yasukazu Nakamura ||, Satoshi Tabata ||, Niels Sandal *, and Jens Stougaard *, **

*Department of Molecular Biology, University of Aarhus, Gustav Wieds Vej 10 and C.F. Møllers Vej Bldg 130, 8000 Aarhus C, Denmark; {dagger}National Food Research Institute, Tsukuba, Ibaraki 305-8642, Japan; {ddagger}John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom; §Center for High Resolution Imaging and Processing, Medical Sciences Institute/Wellcome Trust Biocentre Complex, School of Life Sciences, University of Dundee, Dundee DD1 5EH, United Kingdom; Botanisches Institut I, Justus-Liebig Universität, D-35390 Giessen, Germany; and ||Kazusa DNA Research Institute, Kisarazu, Chiba 292-0818, Japan

Edited by Sharon R. Long, Stanford University, Stanford, CA

Accepted for publication November 14, 2005.

Received for publication October 11, 2005.

Abstract: Nuclear-cytoplasmic partitioning and traffic between cytoplasmic and nuclear compartments are fundamental processes in eukaryotic cells. Nuclear pore complexes mediate transport of proteins, RNAs and ribonucleoprotein particles in and out of the nucleus. Here we present positional cloning of a plant nucleoporin gene, Nup133, essential for a symbiotic signal transduction pathway shared by Rhizobium bacteria and mycorrhizal fungi. Mutation of Nup133 results in a temperature sensitive nodulation deficient phenotype and absence of mycorrhizal colonization. Root nodules developing with reduced frequency at permissive temperatures are ineffective and electron microscopy show that Rhizobium bacteria are not released from infection threads. Measurement of ion fluxes using a calcium-sensitive dye show that Nup133 is required for the Ca2+ spiking normally detectable within minutes after application of purified rhizobial Nod-factor signal molecules to root hairs. Localization of NUP133 in the nuclear envelope of root cells and root hair cells shown with enhanced yellow fluorescent protein fusion proteins suggests a novel role for NUP133 nucleoporins in a rapid nuclear–cytoplasmic communication after host–plant recognition of symbiotic microbes. Our results identify a component of an intriguing signal process requiring interaction at the cell plasma membrane and at intracellular nuclear and plastid organelle-membranes to induce a second messenger.

Key Words: legume symbiosis • nucleoporin • nuclear pore • plant–microbe interaction

Author contributions: T.H.J., S.T., and J.S. designed research; N.K., L.H.M., S.R., M.F., H.M., E.K.J., H.H.F., L.L.H., S.S., Y.N., and N.S. performed research; N.K., L.H.M., S.R., M.F., E.M.H.Q., H.M., J.A.D., E.K.J., H.H.F., L.L.H., S.S., N.S., and J.S. analyzed data; and J.S. wrote the paper.

Conflict of interest statement: No conflicts declared.

This paper was submitted directly (Track II) to the PNAS office.

Abbreviation: eYFP, enhanced yellow fluorescent protein.

Data deposition: The sequences reported in this paper have been deposited in the GenBank database (accession nos. AJ890251 [GenBank] , AJ890252 [GenBank] , and AP008949–AP008952).

** To whom correspondence should be addressed. E-mail: stougaard{at}

© 2006 by The National Academy of Sciences of the USA

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