Scientia Agricultura Sinica ›› 2024, Vol. 57 ›› Issue (14): 2803-2814.doi: 10.3864/j.issn.0578-1752.2024.14.009

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

Influences of Long-Term Appling Different Fertilizers on the Activities and Abundances of Canocial Ammonia Oxidizers and Comammox in Paddy Soil

ZHANG XiaoQin1(), YIN Chang2, LI Zheng1, TANG Xu2, LI Yan2, WU ChunYan2()   

  1. 1 Zhejiang Agricultural Product Green Development Center, Hangzhou 310003
    2 Institute of Environmental, Resources and Soil Fertilizer, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021
  • Received:2023-08-02 Accepted:2024-05-13 Online:2024-07-16 Published:2024-07-24
  • Contact: WU ChunYan

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Abstract:

【Objective】 This work aimed to investigate the impact of long-term applying organic and inorganic fertilizers on the abundance and activities of complete ammonia oxidizers (Comammox), ammonia-oxidizing archaea (AOA) and bacteria (AOB), as well as the abundances of Nitrospira and Nitrobacter in paddy soil, providing a scientific basis for mitigating greenhouse gas emissions and promoting sustainable agricultural production.【Method】 By utilizing multiple specific inhibitors (i.e., acetylene, 1-octyne, and DMPP) in conjunction with real-time quantitative PCR (qPCR), this work examined the differences in activities and abundances of Comammox, AOA, and AOB as well as the abundances of Nitrobacter and Nitrospira in a paddy soil under four fertilization regimes: plots without fertilizer (CK), with manure (M), with chemical fertilizer (NPK), and with combination of manure and chemical fertilizer (MNPK).【Result】 The long-term application of organic fertilizer significantly stimulated the activities of Comammox and AOA (Two-way ANOVA, P<0.001). In those plots solely receiving organic manure, Comammox accounted for as high as 64.2% of total ammonia oxidizing activity, while the inorganic fertilizer application only enhanced the activity of AOB (Two-way ANOVA, P<0.001). qPCR demonstrated that chronic organic amendment significantly increased the abundances of AOA, Nitrospira, Comammox clade A and Clade B (Two-way ANOVA, P<0.001); whereas inorganic amendment increased the abundances of AOB and Nitrobacter (Two-way ANOVA, P<0.001). The correlation analysis revealed there were positive correlations between activities of AOA and Comammox with moisture content, organic matter (OM), total nitrogen (TN), total phosphorus (TP), available phosphorus (AP), alkaline hydrolysable nitrogen (AN), as well as AOA and Nitrospira abundances, while the activity of Comammox was positively correlated with the abundance of Comammox clade A as well. Additionally, the activity of AOB showed positive correlations with AOB and Nitrobacter abundances, nitrate content, and available potassium (AK).【Conclusion】 Comammox played an important role in nitrification of the tested paddy soil, with its abundance and activity primarily influenced by the changes in moisture content, OM, TN, TP, AP, and AN etc..

Key words: Comammox, nitrification activity, population abundance, organic fertilizers, chemical fertilizers, qPCR

Table 1

The theoretical transforming pathways of nitrate and nitrite under different inhibitor treatments"

途径 Pathway N CH DP OC
硝酸盐和亚硝酸消耗
Consumption of nitrate and nitrite		 √ √ √ √
异养硝化 Heterotrophic nitrification √ √ √ √
AOA √ － √ √
AOB √ － － －
Comammox √ － － √

Table 2

The basic soil physic-chemical properties under different fertilization treatments"

处理
Treatment		 pH 含水量Water content
(%)		 硝态氮
含量
NO3--N
(mg·kg-1)		 铵态氮
含量
NH4+-N
(mg·kg-1)		 有机质
含量
Organic matter
(g·kg-1)		 全氮含量Total nitrogen
(g·kg-1)		 全磷含量Total phosphorus
(g·kg-1)		 全钾含量Total potassium
(g·kg-1)		 有效磷
含量Available phosphorus
(mg·kg-1)		 速效钾
含量Available potassium
(mg·kg-1)		 碱解氮含量Alkaline hydrolyzable nitrogen
(mg·kg-1)
CK 6.09b 25.35a 17.61a 18.12a 19.74a 1.24a 0.79a 23.29a 55.39a 64.60a 102.16a
NPK 5.49a 25.71a 63.77b 18.62a 19.47a 1.30a 1.11ac 24.19a 135.71d 88.92b 112.24a
M 6.39c 29.88b 25.08a 26.13b 29.73b 1.88b 1.42bc 24.50a 180.29b 66.65a 149.62b
MNPK 5.84b 29.51b 61.50b 18.17a 34.32c 2.29c 1.86b 24.39a 248.61c 94.55b 172.28c

Fig. 1

The shift in concentrations of NO2- and NO3- of soil samples of different fertilization regimes under different inhibitor treatments"

Fig. 2

The nitrifying activities (A) of AOA, AOB, and Comammox as well as their respective contributions to total autotrophic nitrifying activity (B) under different fertilization regimes"

Table 3

The effects of inorganic and organic fertilization on ammonia oxidizers’ activities and population abundances of related functional guilds analyzed by two-way ANOVA"

项目
Item		 有机肥
Organic fertilization		 无机肥
Inorganic fertilization		 无机肥×有机肥
Organic fertilization× Inorganic fertilization
F1,8 P F1,8 P F1,8 P
AOA活性AOA activity 82.83 <0.001 3.02 0.120 5.48 0.047
AOB活性AOB activity 0.77 0.134 24.89 <0.001 2.78 0.134
Comammox活性Comammox activity 112.82 <0.001 0.56 0.477 0.10 0.760
AOA丰度AOA abundance 85.64 <0.001 14.84 <0.001 10.85 0.002
AOB丰度AOB abundance 23.68 <0.001 1179.28 <0.001 47.10 <0.001
Comammox分枝A丰度ComA abundance 31.62 <0.001 14.15 <0.001 5.65 0.024
Comammox分枝B丰度ComB abundance 36.64 <0.001 11.04 0.002 35.65 <0.001
Nitrobacter丰度Nitrobacter abundance 59.57 <0.001 470.66 <0.001 62.40 <0.001
Nitrospira丰度Nitrospira abundance 187.22 <0.001 0.042 0.840 0.75 0.394

Fig. 3

The inhibitory effect of 1-octyne and C2H2 on the population growth of AOB in NPK and MNPK treatments"

Fig. 4

The abundances of ammonia oxidizers and nitrifiers under different fertilization treatments"

Fig. 5

The Pearson correlation coefficient matrix plot H2O: Water content; OM: Organic matter; TN: Total nitrogen; TP: Total phosphorus; TK: Total potassium; AP: Available phosphorus; AK: Available potassium; AN: Alkaline hydrolyzable nitrogen; AOA: AOA activity; AOB: AOB activity; Comammox: Comammox activity; qAOB: AOB abundance; qAOA: AOA abundance; qComA: Abundance of Comammox clade A; qComB: Abundance of Comammox clade B; qNitrobacter: Nitrobacter abundance; qNitrospira: Nitrospira abundance, respectively. Only correlation coefficient with a significance level P<0.05 was shown"

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