长期施氮对酸性紫色土氨氧化微生物群落及其硝化作用的影响

doi:10.3864/j.issn.0578-1752.2022.03.009

摘要/Abstract

摘要： 目的研究长期施氮对酸性紫色土壤中氨氧化古菌（AOA）和氨氧化细菌（AOB）群落特征的影响,揭示氨氧化微生物群落的驱动因子及其调控硝化作用的微生物学机制。方法依托四川雅安玉米体系施氮长期定位试验（始于2010年）,试验处理包括5个供氮水平,即0（N0）、90（N90）、180（N180）、270（N270）和360（N360）kg N·hm^-2,通过Illumina Miseq高通量测序技术测定AOA和AOB的群落,探究长期施氮对氨氧化微生物群落介导的硝化作用的影响。结果长期施氮影响AOA和AOB的α-多样性（包括丰富度指数和香农-威纳指数）、群落结构和群落组成。其中,随着施氮量的增加,AOA 丰富度指数无显著变化,香农-威纳指数显著降低,AOB 丰富度指数和香农-威纳指数均显著增加;长期不同供氮水平显著影响AOA和AOB的群落结构,供氮水平的增加显著降低AOB优势类群中 Nitrosospira Cluster 3a.1的相对丰度（P<0.05）,同时显著增加了Cluster 3a.2、Cluster 9和Cluster 1的相对丰度（P<0.05）,而对AOA优势类群无显著影响。土壤pH、全氮（TN）、有机质（SOM）、NH₄⁺-N和NO₃^--N均显著影响AOA和AOB的α-多样性,其中,pH与AOB 丰富度指数和香农-威纳指数呈显著负相关（P<0.01）,而与AOA 香农-威纳指数呈显著正相关,而TN、SOM、NH₄⁺-N和NO₃^--N与AOB 丰富度指数和香农-威纳指数呈显著正相关,与AOA 香农-威纳指数呈显著负相关。同时,pH、TN、NO₃^--N 、SOM 和NH₄⁺-N显著影响AOA和AOB的群落结构（P<0.05）。结构方程模型（SEM）的结果表明,长期施氮通过降低土壤pH、提高TN和NO₃^--N含量、改变AOA和AOB的α-多样性和群落结构,进而提高了土壤的硝化势。结论长期施氮通过改变酸性紫色土壤pH、TN、NH₄⁺-N和NO₃^--N和氨氧化微生物的α-多样性和群落结构进而影响了硝化势。

关键词: 酸性紫色土, 供氮水平, 氨氧化细菌, 氨氧化古菌, 硝化势

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^--Nwere 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

邹温馨, 苏卫华, 陈远学, 陈新平, 郎明. 长期施氮对酸性紫色土氨氧化微生物群落及其硝化作用的影响[J]. 中国农业科学, 2022, 55(3): 529-542.

ZOU WenXin, SU WeiHua, CHEN YuanXue, CHEN XinPing, LANG Ming. Effects of Long-Term Nitrogen Application on Ammonia Oxidizer Communities for Nitrification in Acid Purple Soil[J]. Scientia Agricultura Sinica, 2022, 55(3): 529-542.

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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

	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*

	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