The full-length cDNA of LeTIR1 gene was isolated from tomato with EST-based in silico cloning followed by RACE amplification. LeTIR1 contained an open reading frame (ORF) 1872 bp long, encoding 624 amino acid residues. The predicted protein LeTIR1 had one F-box motif and eleven leucine-rich repeats (LRRs), all of which are highly conserved in TIR1 proteins of other plant species. Phylogenetic analysis showed that the LeTIR1 protein shared high similarity with other known TIR1 proteins. Both sequence and phylogenetic analysis suggested that LeTIR1 is a TIR1 homologue and encodes an F-box protein in tomato. Semi-quantitative RT-PCR indicated that LeTIR1 was expressed constitutively in all organs tested, with higher expression in stem than root, leaf, flower and fruit. Its expression level was positively correlated with the auxin distribution in stem or axillary shoot, and was induced by spraying exogenous IAA.
The effects of a microbial inoculant (Thervelics®: a mixture of cells of Bacillus subtilis C-3102 and carrier materials) on rice (Oryza sativa cv. Milkyprincess) and barley (Hordeum vulgare cv. Sachiho Golden) were evaluated in four pot experiments. In the first and second experiments, the dry matter production of rice and barley increased significantly by 10–20% with the inoculation of the mixture at a rate of 107 cfu ⋅ g–1 soil compared with the non-inoculated control. In the third experiment, the growth promoting effects of the mixture, the autoclaved mixture and the carrier materials were compared. The dry mater production of rice grains was the highest in the mixture, and it was significantly higher in the three treatments than in the control, suggesting that the carrier materials may also have a plant growth promoting effect and the living cells might have an additional stimulatory effect. To confirm the efficacy of the living cells in the mixture, only B. subtilis C-3102 cells were used in the fourth experiment. In addition, to estimate the mechanisms in growth promotion by B. subtilis C-3102, three B. subtilis strains with similar or different properties in the production of indole-3-acetic acid (IAA), protease and siderophore and phosphatesolubilizing ability were used as reference strains. Only B. subtilis C-3102 significantly increased the dry matter production of rice grains and the soil protease activity was consistently higher in the soil inoculated with B. subtilis C-3102 throughout the growing period. These results indicate that the microbial inoculant including live B. subtilis C-3102 may have growth promoting effects on rice and barley.