Scientia Agricultura Sinica ›› 2026, Vol. 59 ›› Issue (13): 2878-2893.doi: 10.3864/j.issn.0578-1752.2026.13.010

• SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT • Previous Articles     Next Articles

Effect of Nitrogen Addition on Soil Microorganisms in Leymus secalinus Grassland in Northern Shanxi

JIA HuiLi1,2,3(), ZHU YiChen1,2,3(), CHEN SiXue1,2,3, LIU Miao1,2,3, DONG KuanHu1,2,3, ZHAO Xiang1,2,3, SHI YongHong1,2   

  1. 1 College of Grassland Science, Shanxi Agricultural University, Taigu 030801, Shanxi
    2 Shanxi Key Laboratory of Grassland Ecological Protection and Native Grass Germplasm Innovation, Taigu 030801, Shanxi
    3 Youyu Loess Plateau Grassland Ecosystem National Research Station, Youyu 037200, Shanxi
  • Received:2025-08-18 Accepted:2025-12-10 Online:2026-07-01 Published:2026-07-01

Abstract:

【Objective】This study aimed to investigate the effects of different nitrogen addition levels on soil microbial community structure, diversity and nitrogen cycling functional genes in Leymus secalinus grassland of northern Shanxi, which held the profound significance for understanding nutrient cycling in grassland ecosystems under the context of nitrogen deposition.【Method】This experiment was conducted based on the multi-level nitrogen (N) addition platform established in 2017 at the Youyu Loess Plateau Grassland Ecosystem Research Station, Shanxi Province. Eight treatments were set up: N0, N1, N2, N4, N8, N16, N24, and N32, corresponding to nitrogen application rates of 0, 1, 2, 4, 8, 16, 24, and 32 g N·m-2·a-1, respectively. Utilizing metagenomic technology, soil microorganisms were studied under different levels of nitrogen addition. 【Result】 (1) Nitrogen addition significantly altered the soil bacterial and fungal community structures. It increased the relative abundance of Acidobacteria and Thermodesulfovibrionales in the bacterial community while reducing that of Nitrospirales. In the fungal community, nitrogen addition increased the relative abundance of Ascomycota but decreased that of Mucoromycota and Basidiomycota. (2) Nitrogen addition significantly affected bacterial community structures richness, but had no significant effect on diversity, with no significant impact on the richness and diversity of fungal communities. (3) Microbial differential groups enriched with different levels of nitrogen addition were different. (4) Nitrogen addition significantly increased the relative abundances of nitrification genes amoABC and hao; the relative abundances of denitrification genes nirK and nirS significantly increased under the N4 treatment, while norBC relative abundances significantly increased under N1 and N24 treatments; the relative abundance of dissimilatory nitrate reduction genes nirBD and nrfAH increased significantly under the N16-N32 treatments, while the relative abundance of napAB and narGHI decreased significantly under the N1, N2, N4, N16 and N24 treatments; the relative abundances of nitrogen fixation genes nifDKH and vnfDKGH significantly increased under the N32 treatment (5) With the increase in nitrogen addition levels, soil water content (SWC) and pH decreased significantly, while total carbon (TC) and total nitrogen (TN) showed a trend of first increasing and then decreasing. Similarly, microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN) also exhibited a trend of first increasing and then decreasing. These changes in soil factors significantly affected the composition of the bacterial community structure, among which SWC was the most significant factor. (6) The genes involved in the process of dissimilatory nitrate reduction to ammonium were significantly negatively correlated with SWC and pH. The genes involved in the process of assimilatory nitrate reduction to ammonium were significantly positively correlated with MBC and MBN, nitrogen fixation process genes were significantly positively correlated with SWC, nitrification process genes were significantly negatively correlated with soil moisture, and were significantly positively correlated with NO3--N.【Conclusion】 In summary, nitrogen addition significantly affected the structure of soil microbial communities by influencing soil water content (SWC) and pH, which notably impacted the relative abundances of genes involved in nitrification, dissimilatory nitrate reduction to ammonium, nitrogen fixation, and denitrification processes, thereby altering the progression of soil nitrogen cycling. Soil microorganisms play a pivotal role in nutrient cycling. Further investigating the effects of nitrogen addition on soil microorganisms not only provided a theoretical basis for the rational management of Leymus secalinus grasslands in northern Shanxi but also held significant implications for the restoration of ecosystem functions in these grasslands.

Key words: soil microorganism, nitrogen addition, microbial community structure, nitrogen cycling functional genes, Leymus secalinus grassland in Northern Shanxi, nitrogen deposition

Table 1

Soil physicochemical properties under different nitrogen addition levels (mean±SD)"

处理Treatment pH SWC (%) TN TC C/N
N0 9.71±0.05a 18.75±0.60a 0.1±0.02c 1.54±0.04d 15.28±2.79ab
N1 9.54±0.02ab 16.78±0.69c 0.12±0.01c 2.11±0.06bc 17.54±1.13a
N2 9.41±0.03ab 17.69±0.81b 0.20±0.02a 2.47±0.08ab 12.29±0.83bc
N4 9.10±0.04b 17.44±0.44b 0.18±0.04a 2.62±0.05a 14.60±2.85b
N8 9.11±0.01b 18.87±0.87a 0.17±0.03ab 1.97±0.04c 11.55±1.68c
N16 8.75±0.04bc 13.22±0.22e 0.18±0.01a 2.26±0.04b 12.41±0.46bc
N24 8.42±0.02bc 14.26±0.26d 0.13±0.03bc 1.74±0.05cd 13.47±2.74bc
N32 8.39±0.01c 13.04±0.51e 0.10±0.01c 1.49±0.05d 14.57±0.94b
TP (g·kg-1) NH4+-N (mg·kg-1) NO3--N (mg·kg-1) MBC (mg·kg-1) MBN (mg·kg-1)
N0 0.55±0.03a 90.44±4.23b 7.15±4.03e 115.04±4.92cd 16.21±1.02e
N1 0.49±0.01a 87.07±2.31cd 21.72±1.72d 119.92±2.81d 30.81±2.17d
N2 0.48±0.01a 80.73±3.22c 50.87±2.12b 230.36±5.35b 37.53±0.82c
N4 0.53±0.02a 102.17±5.17bc 44.01±1.27c 174.77±3.22c 45.42±2.40b
N8 0.55±0.02a 133.75±5.24a 21.63±1.53d 412.08±9.94a 69.98±2.98a
N16 0.50±0.03a 131.62±1.42a 29.15±0.85cd 98.37±2.36de 19.04±1.33e
N24 0.55±0.02a 99.64±3.63bc 55.41±1.19b 86.35±1.34e 6.87±0.86f
N32 0.54±0.04a 112.57±4.43b 127.98±4.91a 153.27±3.27cd 18.46±1.43e

Fig. 1

Analysis of community structure of soil bacteria and fungi under different nitrogen addition levels"

Fig. 2

Chao1 index, Shannon index, and Simpson index of soil bacteria and fungi under different nitrogen addition levels Different lowercase letters between treatments indicate significant differences at the significant level of 0.05"

Fig. 3

LefSe analysis of multilevel species differences of soil bacteria and fungi under different nitrogen addition levels (P<0.05)"

Fig. 4

The relative abundance of genes involved in various pathways of nitrogen cycling under different nitrogen addition levels"

Fig. 5

Redundancy analysis of bacteria (a) and fungi (b) under different nitrogen addition levels"

Fig. 6

Relationship between functional genes of N cycle pathway and soil physicochemical properties under different nitrogen addition levels (P<0.05) DNRA: Dissimilatory nitrate reduction to ammonia, ANRA: Assimilatory nitrate reduction to ammonia, Den: Denitrification, NF: Nitrogen fixation, Nit: Nitrification"

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