The SNARE protein SYP71 expressed in vascular tissues is involved in symbiotic nitrogen fixation in nodules

Hakoyama, Tsuneo*; Oi, Ryo*; Hazuma, Kazuya*; Suga, Eri*; Adachi, Yuka*; Kobayashi, Mayumi*; Akai, Rie*; Sato, Shusei*; Fukai, Eigo*; Tabata, Satoshi*; Shibata, Satoshi*; Wu, G.-J.*; Hase, Yoshihiro; Tanaka, Atsushi; Kawaguchi, Masayoshi*; Kochi, Hiroshi*; Umehara, Yosuke*; Suganuma, Norio*

Rhizobial symbiotic nitrogen fixation in root nodules is regulated by the host legume genes. Fix- mutants that exhibit lower or no nitrogen-fixation activity are useful to identify host plant genes required for symbiotic nitrogen fixation. Here, we show a novel Fix- mutant defective of a SNARE protein. The mutant formed nodules that displayed lower nitrogen fixation activity, and the growth of the host plant was retarded. Exogenous combined nitrogen almost recovered the growth of the mutant. Numbers of nodules formed on the mutant were similar to those on the wild-type plant. However, the mutant nodules were smaller and showed early senescence. The causal gene was identified by map-based cloning, and the predicted protein was appeared to be homologous to one of SNARE proteins found in Arabidopsis thaliana. The responsible gene was expressed ubiquitously in shoot, roots and nodules. In roots and nodules, the transcripts were detected in vascular bundles. These results indicated that a SNARE protein expressed in vascular tissues is required for nitrogen fixation activity of rhizobia in nodules.