Plant and Cell Physiology Advance Access originally published online on October 20, 2006
Plant and Cell Physiology 2006 47(11):1572-1581; doi:10.1093/pcp/pcl022
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LjnsRING, a Novel RING Finger Protein, is Required for Symbiotic Interactions Between Mesorhizobium loti and Lotus japonicus
1 Faculty of Agriculture, Kagawa University, Miki, Kita, Kagawa, 761-0795 Japan
2 Division of Plant Sciences, National Institute of Agrobiological Sciences, Tsukuba, Ibaraki, 305-8602 Japan
* Corresponding author: E-mail, kouchih{at}nias.affrc.go.jp; Fax, +81-29-838-8347.
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Abstract |
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Nodule-specific (nodulin) genes are thought to play crucial roles during establishment of the nitrogen-fixing symbiosis between legume plants and Rhizobium bacteria. On the basis of a gene expression database for early stages of the nodulation process of Lotus japonicus, previously constructed by a cDNA macroarray analysis, we identified a novel nodulin gene, LjnsRING, which encodes a protein with a typical RING-H2 finger domain that is well conserved in a number of plant E3 ubiquitin ligases. LjnsRING transcripts were almost exclusively expressed in nodules, and very low expression was detected in roots and shoots. RNA interference (RNAi) knockdown of LjnsRING by hairy root transformation caused impaired root growth together with abortion of nodule formation. Examination with lacZ-labeled Mesorhizobium loti indicated that infection thread formation in the RNAi transgenic hairy roots was significantly inhibited. Analysis using a chimeric gene of LjnsRING promoter and ß-glucuronidase (GUS) coding region demonstrated that LjnsRING transcription in nodules was restricted to the infected cells. These results suggest the requirement for LjnsRING in rhizobial infection and the subsequent nodule formation process.
Keywords: Lotus japonicus - Mesorhizobium loti - Nodulation - RNAi - Agrobacterium rhizogenes
Abbreviations: d.p.i., days post-inoculation; EST, expressed sequence tag; GFP, green fluorescent protein; GUS, ß-glucuronidase; ihpRNA, intron-containing hairpin RNA; IT, infection thread; ORF, open reading frame; RNAi, RNA interference; RT–PCR, reverse transcription–PCR; siRNA, small interfering RNA
The nucleotide sequence of the LjnsRING cDNA reported in this paper has been submitted to DDBJ/EMBL/GENEBANK databases under the accession number, AB272096.
(Received September 2, 2006; Accepted October 8, 2006)
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