Nitrate is the primary form of nitrogen uptake in plants, mainly transported by nitrate transporters (NRTs), including NPF (NITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER FAMILY), NRT2 and NRT3. In this study, we identified a total of 78 NPF, seven NRT2, and two NRT3 genes in maize. Phylogenetic analysis divided the NPF family into eight subgroups (NPF1-NPF8), consistent with the results in Arabidopsis thaliana and rice. The NRT2 family appears to have evolved more conservatively than the NPF family, as NRT2 genes contain fewer introns. The promoters of all NRTs are rich in cis-acting elements responding to biotic and abiotic stresses. The expression of NRTs varies in different tissues and developmental stages, with some NRTs only expressed in specific tissues or developmental stages. RNA-seq analysis using Xu178 revealed differential expression of NRTs in response to nitrogen starvation and nitrate resupply. Moreover, the expression patterns of six key NRTs genes (NPF6.6, NPF6.8, NRT2.1, NRT2.5 and NRT3.1A/B) varied in response to alterations in nitrogen levels across distinct maize inbred lines with different nitrogen uptake rates. This work enhances our understanding of the structure and expression of NRTs genes, and their roles in nitrate response, paving the way for improving maize nitrogen efficiency through molecular breeding.

The overapplication of mineral nitrogen (N) fertilizer has become a scientific problem to be solved in the North China Plain; meanwhile, most organic manure cannot be recycled into the soil. To make full use of organic manure and decrease the applied rate of mineral N fertilizer, a four-year fertilization experiment was carried out to assess the effects of substituting mineral fertilizer with organic manure on soil organic carbon (SOC), labile organic carbon (LOC), stable organic carbon (UOC), microbial biomass carbon (MBC), and soil enzymatic activity. There were six treatments including unfertilized control (CK); mineral fertilizer without organic manure (FR); and mineral nitrogen substituted by organic manure at 20%, 40%, 60%, and 80% (FM1, FM2, FM3, and FM4), respectively. The results showed that SOC, UOC, and LOC were significantly increased by 13.30~40.56%, 41.10~121.92%, and 4.91~15.57% with increasing amounts of organic manure, respectively. Adding organic manure also enhanced urease, catalase, sucrase, and cellulase activities as well as MBC in surface soil (0–20 cm). The present study revealed the activities of urease, catalase, sucrase, and cellulase were closely related to SOC and its components. Following organic manure application, MBC and enzymatic activities had active effects in enhancing LOC and UOC sequestration based on path analyses. Additionally, organic manure increased SOC by direct and indirect enhancement of UOC and LOC, respectively, and improved SOC sequestration as compared to FR treatment. Finally, the study revealed that SOC quantity and quality were both significantly improved by combined application of mineral fertilizer and organic manure, and this effect was enhanced with the increase of organic manure application. Considering the improvements in soil properties and grain yields, substituting mineral fertilizer with organic manure might be a promising alternative in the North China Plain.

The input of exogenous organic materials is an effective way to improve soil organic matter (SOM) content in cropland. The exploration of the impact of new organic materials such as woody peat on black soil fertility can provide an important reference for preventing the degradation of black soil in Northeast China. In this study, the effects of adding woody peat to SOM and the soil-integrated fertility of black cropland were studied by seven treatments (no organic matter addition, CK; crop straw returning, SR; decomposed straw addition, DS; organic manure addition, OM; 6 t/ha woody peat addition, LWP; 10.5 t/ha natural humus material addition, MWP; and 15 t/ha natural humus material addition, HWP). The results show that natural humus material additions (LWP, MWP, and HWP treatments) could significantly increase SOM (increased by 4.79~9.41 g/kg), labile SOM (increased by 2.49~4.52 g/kg), and recalcitrant SOM (increased by 2.13~6.39 g/kg) components, respectively. For comparison, traditional organic material inputs (SR, DS, and OM treatments) had no significant effect on SOM but induced an increase in the labile SOM component in the following year. This study also found that natural humus material additions could improve soil-integrated fertility in a year term, especially in promoting SOM accumulation. However, organic manure amendment showed both the advantage of increasing soil fertility slightly and the disadvantage of increasing soil salt sharply. In conclusion, compared with traditional exogenous organic materials, the natural humus material amendment technique can rapidly increase the total SOM quantity and its different stability components and has a great effect in improving the integrated fertility of black soil. Thus, it is of significance to further study the potential of natural humus material amendment in the fertility of black soil in future.

Bio-solubilization of lignite is a promising technology to transform coal into humic acids (HAs) which are broadly used in agriculture. In this work, HAs were extracted from lignite using the cell-free filtrate (CFF) of Penicillium ortum MJ51. The extraction method was optimized using response surface methodology (RSM) based on the interactive effects of nitric acid concentrations, coal loading ratio, extraction temperature and time as input factors, and the absorbance of HAs at 450 nm wavelength as the output response. Under optimized conditions (lignite pretreated with 4.7 N HNO₃, coal loading ratio of 4.9%, temperature of 77.3 °C and time of 8.6 hours), the absorbance at 450 nm peaked at 70.28, and the concentration and extraction yield of HAs were 31.3 g L⁻¹ and 63.9%, respectively, which were dramatically higher than those observed for traditional biological methods (0.7 g L⁻¹ and 14.1%, respectively). The qualities of HAs produced under optimized conditions were evaluated and compared with those extracted by the conventional chemical method. The optimized process resulted in better HA quality indices, including lower molecular mass; higher nitrogen; less aromatic carbon; more aliphatic and carboxylic carbon; and higher bioactivity for promoting plant growth. Moreover, the anti-flocculation ability was improved, thereby supporting its applicability in agriculture. Extraction of HAs from lignite using the CFF of P. ortum MJ51 provides a novel technological approach for the efficient conversion of lignite to bio-active HAs.