Non-symbiotic hemoglobins (ns-Hbs) are found in all plants, although their physiological function remains to be determined. The present study was undertaken to explore the mode of induction of ns-Hb genes by metabolites of nitrate assimilation using cultured rice (Oryza sativa L.) cells. Two class-1 ns-Hb genes, ORYsa GLB1a and ORYsa GLB1b, were strongly induced by nitrate, nitrite and nitric oxide (NO) donors, S-nitroso-N-acetylpenicillamine and sodium nitroprusside. The rapid and transient accumulation of ORYsa GLB1a and ORYsa GLB1b transcripts in response to nitrate, nitrite and NO donors was similar to that of nia1, which encodes NADH-nitrate reductase (NR), although repression by glutamine and asparagines was significant only for nia1. In the mutants defective in NR mRNA expression, nitrate, nitrite and NO donors failed to induce not only nia1 but also ORYsa GLB1a and ORYsa GLB1b transcripts, indicating that the induction of ns-Hb genes is closely associated with that of the NR gene. Although the kinetics of induction by nitrate, nitrite and NO donors are similar for the two ns-Hb genes, an inhibitor study demonstrated that de novo synthesis of the protein in cytoplasm is essential for inducing ORYsa GLB1b. In contrast, ORYsa GLB1a, like nia1, can be induced in the primary response to these signals without de novo protein synthesis. The role of nitrate, nitrite and NO in the induction of ns-Hb gene expression in rice cells and the possible cellar function of ns-Hbs were discussed in relation to nitrate reduction pathways.

Biochar is widely used as a soil amendment to increase crop yields. However, the impact of the interaction between the biochar and microbial inoculants (e.g., biofertilizer) on plant nutrient uptake and yield in forage rice is not fully understood. A greenhouse study was conducted to evaluate the synergistic effects of rice-husk biochar and Bacillus pumilus strain TUAT-1 biofertilizer application on growth, yield, and nutrient uptake in two forage rice genotypes; Fukuhibiki and the newly bred line, LTAT-29. Positive effects of biochar and biofertilizer, alone or in a combination, on growth traits, nutrient uptake, and yield components were dependent on the rice genotypes. Biochar and TUAT-1 biofertilizer influenced the overall growth of plants positively and increased straw and above-ground biomass in both genotypes. However, although biochar application significantly increased grain yield in LTAT-29, this was not the case in Fukuhibiki. Biochar and TUAT-1 biofertilizer, either alone or combined, significantly affected plant nutrient uptake but the effect largely depended on rice genotype. Results of this study indicate that biochar amendment and TUAT-1 biofertilizer can enhance forage rice productivity depending on genotypes, and therefore, there is a need to consider plant genetic composition when evaluating the potential for crop response to these soil amendments before application on a commercial scale.

Bacillus pumilus strain TUAT1 is a plant growth-promoting bacterium (PGPB) applied as a biofertilizer, containing its spores, for rice. In this study, we analyzed the short-term effects of biofertilization on plant growth in the nursery and long-term effects on plant vegetative growth, yield, and lodging resistance in paddy fields using animal feed rice (‘Fukuhibiki’ and line LTAT-29 which was recently officially registered as a cultivar ‘Monster Nokodai 1′) and fodder rice (line TAT-26). The effects of the biofertilization were analyzed under two nitrogen treatments and at two transplanting distances in the field. The application of 10⁷ colony forming units (CFU) mL⁻¹ bacterial spore solution to seeds on plant box significantly improved the initial growth of rice. The biofertilizer treatment with this strain at 10⁷ CFU g⁻¹ onto seeds in nursery boxes increased the nitrogen uptake at the early growth of rice in the field, resulting in higher growth at the late vegetative growth stage (e.g., tiller number and plant height). Furthermore, the improvement of growth led to increases of not only yield components such as the total panicle number (TPN) and the number of spikelets in a panicle (NSP) in LTAT-29 but also the straw yield and quality of TAT-26. The lodging resistances of these forage rice plants were also improved due to the increased root development and photosynthesis creating tougher culms.