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Plant and Cell Physiology Advance Access published online on January 24, 2005
Plant and Cell Physiology, doi:10.1093/pci/pci001
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Plant and Cell Physiology 2004 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received May 19, 2004
Accepted October 18, 2004

Regular Paper

Symbiotic Rhizobium and Nitric Oxide Induce Gene Expression of Nonsymbiotic Hemoglobin in Lotus japonicus

Yoshikazu Shimoda 1, Maki Nagata 1, Akihiro Suzuki 2, Mikiko Abe 2, Shusei Sato 3, Tomohiko Kato 3, Satoshi Tabata 3, Shiro Higashi 2, and Toshiki Uchiumi 2*

1 Graduate School of Science and Technology, Kagoshima University, Kagoshima, 890-0065 Japan
2 Department of Chemistry and Bioscience, Kagoshima University, Kagoshima, 890-0065 Japan
3 Kazusa DNA Research Institute, Chiba, 292-0812 Japan

* To whom correspondence should be addressed.
Toshiki Uchiumi, E-mail: uttan{at}sci.kagoshima-u.ac.jp


  Abstract

We characterized the expression profiles of LjHb1 and LjHb2, nonsymbiotic hemoglobin (nonsym-Hb) genes of Lotus japonicus. Although LjHb1 and LjHb2 showed 77% homology in their cDNA sequences, LjHb2 is located in a unique position in the phylogenetic tree of plant Hbs. The 5'-upstream regions of both genes contain the motif AAAGGG at a position similar to that in promoters of other nonsym-Hb genes. Expression profiles obtained by using quantitative RT-PCR showed that LjHb1 and LjHb2 were expressed in all tissues of mature plants, and expression was enhanced in mature root nodules. LjHb1 was strongly induced under both hypoxic and cold conditions, and by the application of nitric oxide (NO) donor, whereas LjHb2 was induced only by the application of sucrose. LjHb1 was also induced transiently by the inoculation with the symbiotic rhizobium Mesorhizobium loti MAFF303099. A candidate gene for pathogen-inducible NO synthase (iNOS) of L. japonicus was also induced by inoculation with M. loti MAFF303099. Observations using fluorescence microscopy revealed the induction of LjHb1 expression is corresponded to the generation of NO. These results suggest that nonsym-Hb and NO may have important roles in stress adaptation and in the early stage of legume-rhizobium symbiosis.

Keywords: hypoxia; Lotus japonicus; nitric oxide; nonsymbiotic hemoglobin; plant hormone; rhizobium.
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