Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (3): 529-542.doi: 10.3864/j.issn.0578-1752.2022.03.009

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

Effects of Long-Term Nitrogen Application on Ammonia Oxidizer Communities for Nitrification in Acid Purple Soil

ZOU WenXin1(),SU WeiHua1,CHEN YuanXue2,CHEN XinPing1,LANG Ming1,*()   

  1. 1 College of Resources and Environment, Southwest University/Chongqing Key Laboratory of Efficient Utilization of Soil and Fertilizer Resources, Chongqing 400715
    2 College of Resource Sciences, Sichuan Agricultural University, Chengdu 611130
  • Received:2021-01-06 Accepted:2021-03-10 Online:2022-02-01 Published:2022-02-11
  • Contact: Ming LANG E-mail:zouwx060@163.com;langming@swu.edu.cn

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Abstract:

【Objective】 The purpose of this study was to explore the effects of long-term nitrogen (N) application on the community characteristics of ammonia oxidizing archaea (AOA) and ammonia oxidizing bacteria (AOB) in acid purple soil, and to reveal the driving factors of ammonia oxidizer communities and the microbial mechanism to regulate nitrification.【Method】 Relying on the long-term gradient N fertilization maize field experiment in Sichuan (beginning in 2010), the treatments included five N supply levels: 0 (N0), 90 (N90), 180 (N180), 270 (N270) and 360 (N360) kg N·hm-2. Illumina Miseq high-throughput sequencing technology was used to determine the communities of AOA and AOB, so as to explore the effect of long-term gradient N application on the nitrification mediated by ammonia oxidizer communities. 【Result】 Long-term N application affected the α-diversity (including sobs index and Shannon-Wiener index), community structure and community composition of AOA and AOB. As N application rate increased, the sobs index of AOA did not change significantly, while the Shannon-Wiener index of AOA decreased significantly, however, both the sobs index and Shannon-Wiener index of AOB increased significantly; long-term gradient N fertilization significantly affected the community structure of AOA and AOB. The relative abundance of the dominant AOB groups named Nitrosospira Cluster 3a.1 significantly reduced while Cluster 3a.2, Cluster 9 and Cluster 1 significantly increased with the increased of the N application (P<0.05); there were no obvious rules for the relative abundance of the dominant AOA group named clade A. pH, TN, SOM, NH4+-N and NO3--N all significantly affected the α-diversity of AOA and AOB. pH was significantly negatively correlated with AOB sobs index and Shannon-Wiener index, and significantly positively correlated with AOA Shannon-Wiener index. TN, SOM, NH4+-N and NO3--N were significantly positively correlated with the AOB sobs index and Shannon-Wiener index, and were significantly negatively correlated with the AOA Shannon-Wiener index. Meanwhile, pH, TN, NO3--N, SOM, NH4+-N significantly affected the community structure of AOA and AOB (P<0.05). Moreover, the results of structural equation modeling (SEM) comprehensively showed that long-term gradient N application reduced pH, increased the TN and NO3--N content, and affected the α-diversity and community structure of AOA and AOB, which indirectly increased soil PNR. 【Conclusion】 Long-term N application affected the soil PNR by changing the soil pH, TN, SOM, NH4+-N, NO3--N, the α-diversity and community structure of ammonia oxidizers.

Key words: acid purple soil, gradient nitrogen fertilization, ammonia oxidizing bacteria, ammonia oxidizing archaea, potential nitrification rate

Table 1

Basic physical and chemical properties of different nitrogen treatments"

处理
Treatment		 全氮
TN (g·kg-1)		 有机质
SOM (g·kg-1)		 pH 铵态氮
NH4+-N (mg·kg-1)		 硝态氮
NO3--N (mg·kg-1)
N0 1.56±0.11c 26.7±1.79c 6.6±0.10a 4.26±0.60d 8.20±0.65c
N90 1.95±0.18b 31.6±3.04ab 6.4±0.08b 5.91±0.43c 12.07±0.70b
N180 2.10±0.14a 28.7±1.93bc 5.9±0.10c 6.50±0.46c 12.79±0.65b
N270 2.18±0.16a 34.1±3.00a 5.8±0.10c 8.00±0.63b 15.77±0.77a
N360 2.21±0.20a 33.0±2.21a 5.9±0.05c 10.56±0.75a 16.16±0.80a
数值为平均值±标准差,n=3。数据后的不同小写字母表示施氮处理间差异显著（P<0.05）。N0、N90、N180、N270和N360 分别表示施氮量为 0、90、180、270和360 kg N·hm-2。下同
The value is the mean±standard deviation, n=3. Difference lowercase letters indicate the significant difference of different nitrogen application treatments (P<0.05). N0, N90, N180, N270, N360 indicate N application rate are 0, 90, 180, 270, 360 kg N·hm-2, respectively. The same as below

Fig. 1

Potential nitrification rate under different N treatments The different letters above the bar graph indicate significant differences between treatments (P<0.05)"

Fig. 2

Effects of different nitrogen application rates on AOA, AOB sobs index (A, B) and Shannon-Wiener index (C, D)"

Fig. 3

Principal coordinate analysis (PCoA) plots of AOA (A) and AOB (B) communities with different nitrogen application rates"

Table 2

Pearson correlation analysis between AOA, AOB α-diversity, potential nitrification rate and soil properties"

AOA
Sobs index		 AOB
Sobs index		 AOA
Shannon-Wiener index		 AOB
Shannon-Wiener index		 硝化势
PNR
pH 0.270 -0.743** 0.862** -0.948** -0.312
TN -0.410 0.633* -0.821** 0.857** 0.366
SOM -0.490 0.603* -0.688** 0.557* 0.278
NH4+-N -0.110 0.628* -0.629* 0.701** 0.386
NO3--N -0.150 0.710** -0.803** 0.817** 0.542*

Fig. 4

Phylogenetic tree (A, C) of AOA, AOB amoA gene sequence with relative abundance >1% and community composition (B, D) of AOA and AOB Other means OTU with relative abundance <1%. There is no significant difference in Cluster3c in different nitrogen treatments in the community composition of AOB, and it is not marked in N0, N90, N180, and N270 treatment"

Fig. 5

Redundancy analysis (RDA) of AOA (A) and AOB (B) communities based on OTU matrices with different nitrogen application ratesTN: Total nitrogen, SOM: Soil organic matter, NH4+-N: Ammonium, NO3--N: Nitrate. Significance: * Means 0.01<P<0.05, ** Means 0.001<P<0.01, *** Means P<0.001"

Fig. 6

Correlation ships between community composition of AOA (A), AOB (B) and environment variables AOA, AOB community composition and environmental variables are subjected to spearman correlation analysis, other means OTU with relative abundance <1%.* Means 0.01<P<0.05, ** Means 0.001<P<0.01, *** Means P<0.001"

Table 3

Pearson correlation analysis of AOA, AOB α-diversity and potential nitrification rate"

AOA Sobs index AOB Sobs index AOA Shannon-Wiener Index AOB Shannon-Wiener index
PNR 0.591* 0.512 0.477 0.335

Table 4

Pearson correlation analysis between AOA, AOB dominant groups (relative abundance >1%) and potential nitrification rate"

AOA优势属 AOA dominant groups AOB 优势属 AOB dominant groups
Cladea Cladeb3 Clade Nitrososphaera Cladee Cluster3a.1 Cluster9 Cluster11 Cluster1 Cluster3a.2 Cluster3c
PNR -0.168 0.170 0.223 -0.075 -0.206 0.335 -0.210 0.182 0.196 0.102

Fig. 7

Correlation ships among nitrogen application rate, environmental factors (pH, TN, NO3--N), diversity of AOA and AOB, community structure of AOA and AOB and potential nitrification rate The width of the arrow indicates the strength of the causal effect. The red and blue arrows indicate the positive and negative relationships between the indicators, respectively. The number above the arrow indicates the path coefficient (P<0.05 is a significant path). The solid line and the dashed line represent the significant path and the insignificant path, respectively. The percentage above each indicator represents the R2 value, which is the variance explained ratio of each variable. The final model fits the data well. The model is: χ2=10.28, df=14, CFI=1.000, NFI=0.946, IFI=1.021, RMSEA<0.001"

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