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Plant and Cell Physiology Advance Access published online on May 13, 2006
Plant and Cell Physiology, doi:10.1093/pcp/pcj054
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Plant and Cell Physiology 2006 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received January 21, 2006
Accepted April 28, 2006

Regular Paper

Identification of a Sed5-like SNARE Gene LjSYP32-1 that Contributes to Nodule Tissue Formation of Lotus japonicus

Ha Thu Mai 1, Mika Nomura 2, Kaoru Takegawa 2, Erika Asamizu 3, Syusei Sato 3, Tomohiko Kato 3, Satoshi Tabata 3, and Shigeyuki Tajima 2 *

1 Department of Life Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kita-gun, Kagawa 761-0795, Japan; Institute of Agricultural Genetics, Conhue-Tuliem, Hanoi, Vietnam
2 Department of Life Science, Faculty of Agriculture, Kagawa University, Miki-cho, Kita-gun, Kagawa 761-0795, Japan
3 Kazusa DNA Research Institute, 1532-3 Yana, Kisarazu, Chiba 292-0812, Japan

* To whom correspondence should be addressed.
Shigeyuki Tajima, E-mail: tajima{at}ag.kagawa-u.ac.jp


  Abstract

We identified a Sed5-like clone LjSYP32-1which contributes to nodule tissue formation and plant growth in Lotus japonicus. In L. japonicus EST clone databases of Kazusa DNA Research Institute, another syntaxin-related clone (LjSYP32-2) was also detected and their nucleotide and amino acid sequences are very similar each other.

Real time PCR and promoter analysis indicated that expression of LjSYP32-1 was dominant comparing to LjSYP32-2 in the various plant organs. Promoter analysis and in situ hybridization revealed that LjSYP32-1 expressed significantly in inner cortex cell layer surrounding the infected zone of young nodules and at meristem area of developing lateral root.

To explore function and physiological role of LjSYP32-1 in nodule and other plant organs, stable transformation lines of Lotus japonicus expressing either sense or anti-sense LjSYP32-1 were prepared. The anti-sense plants showed a significant retarding plant growth phenotype, suggesting the role of LjSYP32-1 for supporting plant growth. In the same transgenic lines, the plants were capable to form nodule, but the acetylene reduction activity reduced around 50% per a plant. The nodules were much smaller and some nodules fused each other by sharing the inner cortex. Such irregular nodules were observed in the rate of two times more comparing to those of wild type plant. The data suggest that LjSYP32-1 is contributing to support plant growth and normal nodule tissue differentiation.

Keywords: Lotus japonicus; Legume Nodules; SNARE; Symbiosis.
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