长期施肥对水稻土典型氨氧化菌和全程氨氧化菌种群活性和丰度的影响

doi:10.3864/j.issn.0578-1752.2024.14.009

摘要/Abstract

摘要：

【目的】通过研究长期施用有机肥和无机肥对硝化菌种群活性和丰度的影响，揭示其种群丰度变化与硝化活性之间的关系，为农业生产中减少温室气体排放和实现可持续发展提供科学依据。【方法】使用特异性抑制剂乙炔、1-辛炔和3,4-二甲基吡唑磷酸盐（DMPP）等，结合实时荧光定量PCR技术（qPCR），分析水稻土长期不施肥（CK）、单施化肥（NPK）、单施有机肥（M），以及化肥与有机肥配合施用（MNPK）4个处理的全程氨氧化菌（Comammox）、氨氧化细菌（AOB）、氨氧化古菌（AOA）种群活性和丰度，以及硝化螺菌（Nitrospira）和硝化杆菌（Nitrobacter）种群丰度差异。【结果】长期施用有机肥显著提高了Comammox和AOA的活性（双因素方差分析，P<0.001），在单施有机肥处理中，Comammox活性贡献比例高达64.2%，而施用无机肥仅显著促进了AOB的活性（双因素方差分析，P<0.001）。qPCR分析结果表明，有机肥处理显著提高了AOA、Nitrospira、Comammox分枝A和分枝B种群的丰度（双因素方差分析，P<0.001），而无机肥处理则显著提高了AOB和Nitrobacter的种群丰度（双因素方差分析，P<0.001）。相关性分析表明，AOA和Comammox的活性与土壤含水量、有机质、全氮、全磷、速效磷、碱解氮含量以及AOA和Nitrospira种群丰度显著正相关，Comammox活性还与Comammox分枝A丰度显著正相关；而AOB活性则与AOB和Nitrobacter种群丰度、硝态氮和速效钾含量呈显著正相关。【结论】 Comammox对供试水稻土的硝化作用有着重要贡献，其活性和种群丰度主要受土壤含水量、有机质、全氮、全磷、速效磷和碱解氮等因子变化的影响。

关键词: 全程氨氧化菌, 硝化活性, 种群丰度, 有机肥, 化肥, 实时荧光定量PCR技术

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

张小琴, 尹昌, 李政, 唐旭, 李艳, 吴春艳. 长期施肥对水稻土典型氨氧化菌和全程氨氧化菌种群活性和丰度的影响[J]. 中国农业科学, 2024, 57(14): 2803-2814.

ZHANG XiaoQin, YIN Chang, LI Zheng, TANG Xu, LI Yan, WU ChunYan. Influences of Long-Term Appling Different Fertilizers on the Activities and Abundances of Canocial Ammonia Oxidizers and Comammox in Paddy Soil[J]. Scientia Agricultura Sinica, 2024, 57(14): 2803-2814.

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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	√	－	－	√

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

项目 Item	有机肥 Organic fertilization		无机肥 Inorganic fertilization		无机肥×有机肥 Organic fertilization× Inorganic fertilization
项目 Item	F_1,8	P	F_1,8	P	F_1,8	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