Chemical fertilizer reduction combined with novel and green agricultural inputs has become an important practice to improve microecological health in agricultural production. Given the close linkages between rhizosphere processes and plant nutrition and productivity, understanding how fertilization impacts this critical zone is highly important for optimizing plant–soil interactions and crop fitness for agricultural sustainability. Here, by using a pot experimental system, we demonstrated that nitrogen fertilizer reduction and microbial agent application promoted plant fitness and altered the microbial community structure in the rhizosphere soil with the following treatments: no fertilization, CK; conventional chemical fertilizer, CF; 30% reduced nitrogen fertilizer, N; 30% reduced nitrogen fertilizer with pure γ-PGA, PGA; 30% reduced nitrogen fertilizer with Bacillus subtilis A-5, A5; 30% reduced nitrogen fertilizer with γ-PGA fermentation broth, FJY. The PGA, A5, and FJY treatments all significantly promoted crop growth, and the FJY treatment showed the strongest positive effect on Chinese cabbage yield (26,385.09 kg/hm²) (P < 0.05). Microbial agents affected the α diversity of the rhizosphere bacterial community; the addition of B. subtilis A-5 (A5 and FJY treatments) significantly affected rhizospheric bacterial community structure. Urease activity and soil pH were the key factors affecting bacterial community structure and composition. The FJY treatment seemed to influence the relative abundances of important bacterial taxa related to metabolite degradation, predation, and nitrogen cycling. This discovery provides insight into the mechanism underlying the effects of microbial agent inputs on rhizosphere microbial community assembly and highlights a promising direction for the manipulation of the rhizosphere microbiome to yield beneficial outcomes.

Bacillus subtilis A-5 has the capabilities of high-molecular-weight γ-PGA production, antagonism to plant pathogenic fungi, and salt/alkaline tolerance. This multifunctional bacterium has great potential for enhancing soil fertility and plant security in agricultural ecosystem. The genome size of B. subtilis A-5 was 4,190,775 bp, containing 1 Chr and 2 plasmids (pA and pB) with 43.37% guanine-cytosine content and 4605 coding sequences. The γ-PGA synthase gene cluster was predicted to consist of pgsBCA and factor (pgsE). The γ-PGA-degrading enzymes were mainly pgdS, GGT, and cwlO. Nine gene clusters producing secondary metabolite substances, namely, four unknown function gene clusters and five antibiotic synthesis gene clusters (surfactin, fengycin, bacillibactin, subtilosin_A, and bacilysin), were predicted in the genome of B. subtilis A-5 using antiSMASH. In addition, B. subtilis A-5 contained genes related to carbohydrate and protein decomposition, proline synthesis, pyruvate kinase, and stress-resistant proteins. This affords significant insights into the survival and application of B. subtilis A-5 in adverse agricultural environmental conditions.

The current fertilizer recommendations for melon plantation have many limitations and exhibit deficiencies regarding future development. Therefore, in this study, the optimal quantities of fertilizer, in terms of the effects of single factors and interaction effects, are studied. There were significant interaction effects between N and P, N and K, P and K; the contents of soluble protein, vitamin C (Vc), and soluble sugar in melon could be improved using the optimal fertilization ratios. The optimal ratio of N:P:K was 2.33:1:3.85, with the amounts of N, P₂O₅, and K₂O, respectively, being 157.5, 67.58, and 260.38 kg/hm², yielding 8.73 g/kg of soluble protein in melon. The optimal ratio of N:P:K was 2.03:1:3.36, with amounts of N, P₂O₅, and K₂O being 157.50, 77.40, and 260.38 kg/hm², respectively, yielding 25.32 g/kg Vc content in melon. Finally, the optimal ratio of N:P:K was 1.53:1:3.36, with the amounts of N, P₂O₅, and K₂O being 118.07, 77.40, respectively, and 260.38 kg/hm², yielding 13.34% soluble sugar content in melon.

The ecological rice–eel co-culture system is not only beneficial for enhancing productivity and sustainability in agriculture but also plays a crucial role in promoting environmental health. In the present study, based on the long-term positioning trial of the rice–eel co-culture system that began in 2016 and was sampled in 2023, the effects of reduced nitrogen fertilizer application on soil physico-chemical properties and the bacterial community were investigated. Treatments included a conventional regular fertilization treatment (RT), rice–eel co-culture system regular fertilization (IT), and nitrogen-reduction 10%, 30%, and 50% fertilization treatments (IT90, IT70, and IT50). Our research demonstrated the following: (1) Compared to RT, IT significantly increased soil water-stable macroaggregates (R0.25), mean weight diameter (MWD), geometric mean diameter (GMD), and available phosphorus content, with the increases of 15.66%, 25.49%, 36.00%, and 18.42%, respectively. Among the nitrogen-reduction fertilization treatments, IT90 showed the most significant effect. Compared to IT, IT90 significantly increased R0.25, MWD, GMD, and available nitrogen content, with increases of 4.4%, 7.81%, 8.82%, and 28.89%, respectively. (2) Compared to RT, at the phylum level, the diversity of Chloroflexi was significantly increased under IT and IT50, and the diversity of Gemmatimonadota was significantly increased under IT90, IT70, and IT50. The diversity of Acidobacteriota was significantly higher in IT90 and IT70 compared to IT. It was shown that the rice–eel co-culture system and nitrogen fertilizer reduction could effectively improve the degradation capacity of organic matter and promote soil nitrogen cycling. In addition, redundancy analysis (RDA) identified total phosphorus, total nitrogen, and available nitrogen (p = 0.007) as the three most important environmental factors driving changes in the bacterial community. (3) The functional prediction analysis of soil microbiota showed that, compared to RT, the diversity of pathways related to biosynthesis (carbohydrate biosynthesis and cell structure biosynthesis) and metabolism (L-glutamate and L-glutamine biosynthesis) was significantly higher under IT70, IT90, IT, and IT50 (in descending order). However, the diversity of pathways associated with degradation/utilization/assimilation (secondary metabolite degradation and amine and polyamine degradation) was significantly lower under all the rice–eel co-culture treatments. In conclusion, the rice–eel co-culture system improved soil physicochemical properties and the soil microbial environment compared with conventional planting, and the best soil improvement was achieved with 10% less N fertilizer application.