The use of nitrogen-fixing bacteria in agriculture is increasingly recognized as a sustainable method to boost crop yields, reduce chemical fertilizer use, and improve soil health. However, the microbial mechanisms by which inoculation with nitrogen-fixing bacteria enhance rice production remain unclear. In this study, rice seedlings were inoculated with the nitrogen-fixing bacterium R3 (Herbaspirillum) at the rhizosphere during the seedling stage in a pot experiment using paddy soil. We investigated the effects of such inoculation on nutrient content in the rhizosphere soil, plant growth, and the nitrogen-fixing microbial communities within the rhizosphere and endorhizosphere. The findings showed that inoculation with the R3 strain considerably increased the amounts of nitrate nitrogen, ammonium nitrogen, and available phosphorus in the rhizosphere by 14.77%, 27.83%, and 22.67%, respectively, in comparison to the control (CK). Additionally, the theoretical yield of rice was enhanced by 8.81% due to this inoculation, primarily through a 10.24% increase in the effective number of rice panicles and a 4.14% increase in the seed setting rate. Further analysis revealed that the structure of the native nitrogen-fixing microbial communities within the rhizosphere and endorhizosphere were altered by inoculation with the R3 strain, significantly increasing the α-diversity of the communities. The relative abundance of key nitrogen-fixing genera such asRalstonia,Azotobacter,Geobacter,Streptomyces, andPseudomonaswere increased, enhancing the quantity and community stability of the nitrogen-fixing community. Consequently, the nitrogen-fixing capacity and sustained activity of the microbial community in the rhizosphere soil were strengthened. Additionally, the expression levels of the nitrogen absorption and transport-related genesOsNRT1andOsPTR9in rice roots were upregulated by inoculation with the R3 strain, potentially contributing to the increased rice yield. Our study has revealed the potential microbial mechanisms through which inoculation with nitrogen-fixing bacteria enhances rice yield. This finding provides a scientific basis for subsequent agricultural practices and is of critical importance for increasing rice production and enhancing the ecosystem services of rice fields.

As core components of ABA signaling pathway, SnRK2s (Sucrose nonfermenting1–Related protein Kinase 2) bind to and phosphorylate AREB/ABF (ABA responsive element binding protein/ABRE-binding factor) transcriptional factors, particularly bZIPs (basic region-leucine zipper), to participate in various biological processes, including flowering. Rice contains 10 SnRK2 members denoted as SAPK1-10 (Stress-Activated Protein Kinase) and dozens of bZIPs. However, which of the SAPKs and bZIPs pair and involve in ABA signaling remains largely unknown. In this study, we carried out a systematical protein-protein interactomic analysis of 10 SAPKs and 9 ABA-inducible bZIPs using yeast-two-hybrid technique, and identified 14 positive interactions. The reliability of Y2H work was verified by in vitro pull-down assay of the key flowering regulator bZIP77 with SAPK9 and SAPK10, respectively. Moreover, SAPK10 could phosphorylate bZIP77 in vitro. Over-expression of SAPK10 resulted in earlier flowering time, at least partially through regulating the FAC-MADS15 pathway. Conclusively, our results provided an overall view of the SAPK-bZIP interactions, and shed novel lights on the mechanisms of ABA-regulated rice flowering.

Abscisic acid (ABA) is a key phytohormone regulating plant development and stress response. The signal transduction of ABA largely relies on protein phosphorylation. However; little is known about the phosphorylation events occurring during ABA signaling in rice thus far. By employing a label-free; MS (Mass Spectrometry)-based phosphoproteomic approach; we identified 2271 phosphosites of young rice seedlings and their intensity dynamics in response to ABA; during which 1060 proteins were found to be differentially phosphorylated. Western-blot analysis verified the differential phosphorylation pattern of D1, SMG1 and SAPK9 as indicated by the MS result; suggesting the high reliability of our phosphoproteomic data. The DP (differentially phosphorylated) proteins are extensively involved in ABA as well as other hormone signaling pathways. It is suggested that ABA antagonistically regulates brassinosteroid (BR) signaling via inhibiting BR receptor activity. The result of this study not only expanded our knowledge of rice phosphoproteome, but also shed more light on the pattern of protein phosphorylation in ABA signaling.

Soil cadmium (Cd) contamination has become a serious problem in China. In this study, the goals of the field experiments were to understand how the application of nitrogen fertilizer at the full heading stage and filling stage reduced the uptake of Cd in rice by affecting the distribution of Cd in iron plaques on the root surfaces and subcellular in the root and flag leaf. The hydroponic culture experiments aimed to explore the effect of interaction or deficiency of nitrogen and cadmium on cadmium accumulation in rice at the late growth stage. The results showed that under the condition of having enough nitrogen supply during the early growth stage, applying nitrogen fertilizer during the full heading stage and filling stage resulted in a significant increase in the concentration of Fe and Cd on the root plaques at the milk stage and mature stage. Additionally, it increased the concentration and proportion of Cd in the soluble fraction of subcellular in the flag leaves at the milk stage. On the other hand, when there was a deficiency of nitrogen, the concentration of Fe in the root plaques increased significantly, while the concentration of Cd in the root plaques decreased significantly. Moreover, the proportion of Cd in the flag leaf cell walls increased significantly. Regardless of whether there was sufficient or deficient nitrogen supply during the early growth stage, applying nitrogen at the full heading stage reduced the Cd concentration in brown rice by 35.11% and 57.25%, respectively. Under hydroponic culture conditions with both Cd exposure and later-stage nitrogen supply significantly increased the Cd concentration in brown rice. However, in the absence of Cd, nitrogen supply significantly reduced the Cd concentration in brown rice. The accumulation of Cd in brown rice was significantly correlated with the accumulation of Fe and Mn at the late growth stage. Our findings indicate that when there is sufficient nitrogen nutrition during the early growth stage of rice, topdressing nitrogen fertilizer at the full heading stage combined with lime with mineral element fertilizers such as Fe and Mn is an effective approach to reduce the presence of cadmium in rice grains.