长期施用氮肥对黑土和风沙土根际微生物群落结构及功能的影响

doi:10.3864/j.issn.0578-1752.2025.03.009

摘要/Abstract

摘要：

【目的】探究氮肥施用对相同气候条件下黑土和风沙土根际微生物的影响，明确群落结构、网络关键物种及指示物种差异响应情况，为指导精准施肥和绿色生产提供科学依据。【方法】依托于吉林省梨树县玉米连作体系氮肥施用的长期定位试验（12年），设置3个氮肥水平0（N0）、168 kgN·hm^-2（N168）和312 kgN·hm^-2（N312），利用高通量测序技术，探究黑土和风沙土根际微生物组成、结构及功能的差异。【结果】长期氮肥施用显著降低了黑土和风沙土根际微生物的Alpha多样性并改变了群落结构，在黑土和风沙土中均为N312处理影响最大，显著降低了2.6%—7.5%的Alpha多样性，整体表现为氮肥施用对风沙土的影响大于黑土；物种分析结果显示，氮肥施用显著增加了拟杆菌门（Bacteroidetes）和Patescibacteria菌门的相对丰度，显著降低了厚壁菌门（Frimicutes）和绿弯菌门（Chloroflexi）的相对丰度，在黑土和风沙土中均为N312处理影响最大（80%—90%）；共现性网络分析发现，氮肥施用对风沙土网络结构的影响大于黑土，同时显著影响了黑土中43%的关键物种和风沙土中全部的关键物种；随机森林的计算结果表明，氮肥施用对风沙土中指示物种的影响大于黑土，与N0相比，N168和N312处理在黑土中无特异性指示物种，而在风沙土中含有两个特异性指示物种，分别为隶属于放线菌门的Intrasporangiaceae科和变形菌门的Noviherbaspirillum属；PICRUSt2功能预测发现，氮肥施用显著影响黑土中88.5%和风沙土中96.2%的氮转化相关功能基因，均为氮肥施用量越高影响越大。【结论】氮肥施用显著降低了根际微生物群落多样性，改变群落结构及物种组成特征，造成了氮转化相关功能基因的显著差异。氮肥的施用对风沙土根际微生物整体的影响大于黑土，且施用量越高影响越大。因此，在黑土和风沙土中进一步推进氮肥减量对维持农田根际微生物群落结构稳定具有重要意义。

关键词: 氮肥施用量, 黑土, 风沙土, 微生物群落, 指示物种, 关键物种

Abstract:

【Objective】This study investigated the differential responses of rhizosphere microbial communities, keystones and indicators to nitrogen fertilizer application in black and sandy soils under identical climatic conditions. The aim of this study was to provide a scientific basis for guiding precision fertilization and promoting green production. 【Method】This study was based on a long-term field experiment (12 years) involving nitrogen fertilizer application in a maize continuous cropping system in Jilin Province. The experimental design included two main treatments: sandy soil and black soil. Under each main treatment, three nitrogen levels were applied: 0 (N0), 168 kgN·hm^-2 (N168), and 312 kgN·hm^-2 (N312). Utilizing high-throughput sequencing technology, the differential impacts of long-term nitrogen fertilizer application on the composition, structure, and functional attributes of rhizosphere microbe communities in both black soil and sandy soil were studied. 【Result】Long-term nitrogen fertilizer application significantly decreased the Alpha diversity and changed the community structure of rhizosphere microbes in both black and sandy soils. The greatest impact was observed under N312 treatment, which significantly reduced Alpha diversity by 2.6%-7.5%. The impact of the same nitrogen application on the rhizosphere microbes was more pronounced in sandy soil than in black soil. Species analysis indicated that nitrogen fertilizer application significantly increased the relative abundance of Bacteroidetes and Patescibacteria phylum and decreased the relative abundance of Frimicutes and Chloroflexi, with the N312 treatment having the greatest impact (80%-90%) in both black and sandy soils. Co-occurrence network analysis revealed that the impact of nitrogen fertilizer application on the network structure was greater in sandy soil than in black soil. Moreover, nitrogen fertilizer application significantly influenced 43% of the keystone species in black soil and all keystone species in sandy soil. Random forest analysis indicated that the impact of nitrogen fertilizer application on indicators was more pronounced in sandy soil than in black soil. Compared with N0, the N168 and N312 treatments had no specific indicator species in black soils, whereas two specific indicators were identified under these two treatments in sandy soil, belonging to the Intrasporangiaceae family of the Actinomycetes phylum and the Noviherbaspirillum genus of the Proteobacteria phylum. PICRUSt2 functional prediction revealed that nitrogen fertilizer application significantly affected 88.5% of nitrogen transformation-related functional genes in black soil and 96.2% in sandy soil, with a greater influence observed at higher nitrogen application rates. 【Conclusion】The research findings suggested that nitrogen fertilizer application significantly reduced the diversity of rhizosphere microbial communities, and changed the community structure and species composition characteristics, resulting in significant differences in nitrogen transformation-related functional genes. The overall impact of nitrogen fertilizer application on rhizosphere microbes was more pronounced in sandy soil than in black soil, with a greater influence observed at higher nitrogen application rates. Therefore, it was important to further promote N fertilizer reduction in black and sandy soils to maintain the stability of rhizosphere microbial community structure in farmland.

Key words: nitrogen fertilizer application, black soil, sandy soil, microbial community, indicator, keystone

王钊, 张冰, 董思奇, 胡雨翕, 齐书宇, 冯国忠, 高强, 周雪. 长期施用氮肥对黑土和风沙土根际微生物群落结构及功能的影响[J]. 中国农业科学, 2025, 58(3): 520-536.

WANG Zhao, ZHANG Bing, DONG SiQi, HU YuXi, QI ShuYu, FENG GuoZhong, GAO Qiang, ZHOU Xue. Effects of Long-Term Nitrogen Fertilizer Application on the Rhizosphere Microbial Community Structure and Function in Black Soil and Sandy Soil[J]. Scientia Agricultura Sinica, 2025, 58(3): 520-536.

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参考文献 58

[1]	张延慧, 刘宇, 韩莹, 董馨宇, 郭探文, 闫秋艳, 闫双堆. 不同比例硫酸铵替代尿素对玉米根际土壤环境及微生物群落的影响. 环境科学, 2023(1): 1-17.
	ZHANG Y H, LIU Y, HAN Y, DONG X Y, GUO T W, YAN Q Y, YAN S D. Effects of different proportions of ammonium sulfate replacing urea on soil nutrients and rhizosphere microbial communities. Environmental Science, 2023(1): 1-17. (in Chinese)
[2]	张淼, 刘俊杰, 刘株秀, 顾海东, 李禄军, 金剑, 刘晓冰, 王光华. 黑土区农田土壤氮循环关键过程微生物基因丰度的分布特征. 土壤学报, 2022, 59(5): 1258-1269.
	ZHANG M, LIU J J, LIU Z X, GU H D, LI L J, JIN J, LIU X B, WANG G H. Distribution characteristics of microbial gene abundance in key processes of soil nitrogen cycling in black soil zone. Acta Pedologica Sinica, 2022, 59(5): 1258-1269. (in Chinese)
[3]	李春玲, 李国山, 于亦忠, 毛文明, 马林山. 减氮施肥及氮肥添加脲酶抑制剂对凉州灌区春玉米产量和氮肥利用的影响. 干旱地区农业研究, 2021, 39(2): 31-36.
	LI C L, LI G S, YU Y Z, MAO W M, MA L S. Effects of reduced nitrogen fertilization and urease inhibitor on the yield and nitrogen use efficiency of spring maize in irrigation areas of Liangzhou. Agricultural Research in the Arid Areas, 2021, 39(2): 31-36. (in Chinese)
[4]	PENG J J, OLADELE O, SONG X J, JU X T, JIA Z J, HU H W, LIU X J, BEI S K, GE A H, ZHANG L M, CUI Z L. Opportunities and approaches for manipulating soil-plant microbiomes for effective crop nitrogen use in agroecosystems. Frontiers of Agricultural Science and Engineering, 2022: 9(3): 333-343. doi: 10.15302/J-FASE-2022450
[5]	GRIFFITH D R, KLADIVKO E J, MANNERING J V, WEST T D, PARSONS S D. Long-term tillage and rotation effects on corn growth and yield on high and low organic matter, poorly drained soils. Agronomy Journal, 1988, 80(4): 599-605.
[6]	尹昌. 农田土壤N₂O产生和还原及相关功能微生物对温度及施肥的响应[D]. 北京: 中国农业科学院, 2017.
	YIN C. Responses of N₂O emission and reduction as well as associated microbes to temperature and different fertilization regimes in arable soils[D]. Beijing: Chinese Academy of Agricultural Sciences, 2017. (in Chinese)
[7]	杨小东. 长期施肥对三种土壤类型酶活性及硝化微生物丰度的影响[D]. 北京: 中国农业科学院, 2020.
	YANG X D. Changes of enzyme activities and abundance of nitrifying microorganism of three diffrent type soils under long-term fertilization[D]. Beijing: Chinese Academy of Agricultural Sciences, 2020. (in Chinese)
[8]	刘杏兰, 高宗, 刘存寿, 司立征. 有机—无机肥配施的增产效应及对土壤肥力影响的定位研究. 土壤学报, 1996, 33(2): 138-147.
	LIU X L, GAO Z, LIU C S, SI L Z. Study on the effect of combined application of organic and inorganic fertilizers on yield increase and its influence on soil fertility. Acta Pedologica Sinica, 1996, 33(2): 138-147. (in Chinese)
[9]	白震, 张旭东, 何红波, 闫颖, 侯松嵋, 陈盈, 解宏图. 长期氮肥施用对农田黑土NLFA与PLFA特性的影响. 土壤学报, 2007, 44(4): 709-716.
	BAI Z, ZHANG X D, HE H B, YAN Y, HOU S M, CHEN Y, XIE H T. Effects of long-term nitrogen fertilizer application on NLFA and PLFA in mollisol farmland. Acta Pedologica Sinica, 2007, 44(4): 709-716. (in Chinese)
[10]	ROUSK J, BÅÅTH E, BROOKES P C, LAUBER C L, LOZUPONE C, CAPORASO J G, KNIGHT R, FIERER N. Soil bacterial and fungal communities across a pH gradient in an arable soil. The ISME Journal, 2010, 4(10): 1340-1351.
[11]	JONES R T, ROBESON M S, LAUBER C L, HAMADY M, KNIGHT R, FIERER N. A comprehensive survey of soil acidobacterial diversity using pyrosequencing and clone library analyses. The ISME Journal, 2009, 3(4): 442-453.
[12]	朱瑞芬, 刘杰淋, 王建丽, 韩微波, 申忠宝, 辛晓平. 基于分子生态学网络分析松嫩退化草地土壤微生物群落对施氮的响应. 中国农业科学, 2020, 53(13): 2637-2646. doi: 10.3864/j.issn.0578-1752.2020.13.012.
	ZHU R F, LIU J L, WANG J L, HAN W B, SHEN Z B, XIN X P. Molecular ecological network analyses revealing the effects of nitrogen application on soil microbial community in the degraded grasslands. Scientia Agricultura Sinica, 2020, 53(13): 2637-2646. doi: 10.3864/j.issn.0578-1752.2020.13.012. (in Chinese)
[13]	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. 中国农业科学, 2022, 55(3): 529-542. doi: 10.3864/j.issn.0578-1752.2022.03.009.
	ZOU W X, SU W H, CHEN Y X, CHEN X P, LANG M. Effects of long-term nitrogen application on ammonia oxidizer communities for nitrification in acid purple soil. Scientia Agricultura Sinica, 2022, 55(3): 529-542. doi: 10.3864/j.issn.0578-1752.2022.03.009. (in Chinese)
[14]	李伊凡, 王俊伟, 陈永豪, 何柄枚, 拉琼. 西藏岗巴拉山土壤细菌多样性海拔分布格局及其驱动因子. 生态学报, 2024, 44(2): 712-722.
	LI Y F, WANG J W, CHEN Y H, HE B M, LA Q. Elevation distribution pattern and driving factors of soil bacterial diversity on Mount Gangbala, Xizang, China. Acta Ecologica Sinica, 2024, 44(2): 712-722. (in Chinese)
[15]	OUYANG Y, EVANS S E, FRIESEN M L, TIEMANN L K. Effect of nitrogen fertilization on the abundance of nitrogen cycling genes in agricultural soils: a meta-analysis of field studies. Soil Biology and Biochemistry, 2018, 127: 71-78.
[16]	徐瑞富, 刘起丽, 王一涵. 玉米不同生育期微生物群落研究. 河南科技学院学报(自然科学版), 2010, 38(2): 11-15.
	XU R F, LIU Q L, WANG Y H. Studies on microbe population of the surface and in vivo of different period of maize. Journal of Henan Institute of Science and Technology (Natural Science Edition), 2010, 38(2): 11-15. (in Chinese)
[17]	周元, 陈远学, 蒋帆, 胡月秋, 龙玲, 裴丽珍, 李建兵, 徐开未. 玉米地土壤微生物量碳、氮及微生物熵对不同物料还田的响应. 水土保持学报, 2020, 34(2): 173-180.
	ZHOU Y, CHEN Y X, JIANG F, HU Y Q, LONG L, PEI L Z, LI J B, XU K W. Responses of soil microbial biomass carbon, nitrogen and microbial entropy to different materials returned to corn fields. Journal of Soil and Water Conservation, 2020, 34(2): 173-180. (in Chinese)
[18]	CHEN S F, ZHOU Y Q, CHEN Y R, GU J. Fastp: an ultra-fast all-in-one FASTQ preprocessor. Bioinformatics, 2018, 34(17): i884-i890.
[19]	MAGOČ T, SALZBERG S L. FLASH: fast length adjustment of short reads to improve genome assemblies. Bioinformatics, 2011, 27(21): 2957-2963. doi: 10.1093/bioinformatics/btr507 pmid: 21903629
[20]	EDGAR R C. UPARSE: highly accurate OTU sequences from microbial amplicon reads. Nature Methods, 2013, 10: 996-998. doi: 10.1038/nmeth.2604 pmid: 23955772
[21]	STACKEBRANDT E, GOEBEL B M. Taxonomic note: a place for DNA-DNA reassociation and 16S rRNA sequence analysis in the present species definition in bacteriology. International Journal of Systematic and Evolutionary Microbiology, 1994, 44(4): 846-849.
[22]	LI S F, HUANG X B, SHEN J Y, XU F D, SU J R. Effects of plant diversity and soil properties on soil fungal community structure with secondary succession in the Pinus yunnanensis forest. Geoderma, 2020, 379: 114646.
[23]	MONTAGNA M, BERRUTI A, BIANCIOTTO V, CREMONESI P, GIANNICO R, GUSMEROLI F, LUMINI E, PIERCE S, PIZZI F, TURRI F, GANDINI G. Differential biodiversity responses between Kingdoms (plants, fungi, bacteria and Metazoa) along an Alpine succession gradient. Molecular Ecology, 2018, 27(18): 3671-3685. doi: 10.1111/mec.14817 pmid: 30146795
[24]	张甜娜, 陈召莹, 张紫薇, 周石磊, 孟佳靖, 陈哲, 张一凡, 董宛佳, 崔建升. 白洋淀冬季沉积物好氧反硝化菌垂向分布特征及群落构建. 环境科学, 2022, 43(5): 2614-2623.
	ZHANG T N, CHEN Z Y, ZHANG Z W, ZHOU S L, MENG J J, CHEN Z, ZHANG Y F, DONG W J, CUI J S. Vertical distribution characteristics and community construction of aerobic denitrification bacteria from the sediments of Baiyangdian Lake during the winter freezing period. Environmental Science, 2022, 43(5): 2614-2623. (in Chinese)
[25]	ZHANG J Y, LIU Y X, ZHANG N, HU B, JIN T, XU H R, QIN Y, YAN P X, ZHANG X N, GUO X X, HUI J, CAO S Y, WANG X, WANG C, WANG H, QU B Y, FAN G Y, YUAN L X, GARRIDO-OTER R, CHU C C, BAI Y. NRT1.1B is associated with root microbiota composition and nitrogen use in field-grown rice. Nature Biotechnology, 2019, 37(6): 676-684. doi: 10.1038/s41587-019-0104-4 pmid: 31036930
[26]	于海玲. 施氮量对土壤微生物群落组成特征的影响研究[D]. 长春: 吉林农业大学, 2017.
	YU H L. Effect of nitrogen application on composition characters of soil microbial communities[D]. Changchun: Jilin Agricultural University, 2017. (in Chinese)
[27]	康巍巍, 王庆贵, 邢亚娟. 氮添加对土壤和根际微生物群落结构影响的研究进展. 世界生态学, 2022, 11(2): 180-193.
	KANG W W, WANG Q G, XING Y J. Research progress on the effects of nitrogen addition on soil and rhizosphere microbial community structure. International Journal of Ecology, 2022, 11(2): 180-193. (in Chinese)
[28]	GEISSELER D, LAZICKI P A, SCOW K M. Mineral nitrogen input decreases microbial biomass in soils under grasslands but not annual crops. Applied Soil Ecology, 2016, 106: 1-10.
[29]	陈国峰, 姬强, 王亚麒, 王锐, 雷金银, 何进勤. 种植模式对宁南山区土壤团聚体和微生物多样性的影响. 中国土壤与肥料, 2023(9): 59-67.
	CHEN G F, JI Q, WANG Y Q, WANG R, LEI J Y, HE J Q. Impact of planting mode on soil aggregates and the microbial diversity in Ningnan mountainous areas. Soil and Fertilizer Sciences in China, 2023(9): 59-67. (in Chinese)
[30]	WANG C, LU X K, MORI T, MAO Q G, ZHOU K J, ZHOU G Y, NIE Y X, MO J M. Responses of soil microbial community to continuous experimental nitrogen additions for 13 years in a nitrogen-rich tropical forest. Soil Biology and Biochemistry, 2018, 121: 103-112.
[31]	ZHANG J Y, AI Z M, LIANG C T, WANG G L, XUE S. Response of soil microbial communities and nitrogen thresholds of Bothriochloa ischaemum to short-term nitrogen addition on the Loess Plateau. Geoderma, 2017, 308: 112-119.
[32]	DJAJADI, ABBOTT L K, HINZ C. Synergistic impacts of clay and organic matter on structural and biological properties of a sandy soil. Geoderma, 2012, 183/184: 19-24.
[33]	DA SILVA K R A, SALLES J F, SELDIN L, VAN ELSAS J D. Application of a novel Paenibacillus-specific PCR-DGGE method and sequence analysis to assess the diversity of Paenibacillus spp. in the maize rhizosphere. Journal of Microbiological Methods, 2003, 54(2): 213-231.
[34]	ZHAO Z B, HE J Z, GEISEN S, HAN L L, WANG J T, SHEN J P, WEI W X, FANG Y T, LI P P, ZHANG L M. Protist communities are more sensitive to nitrogen fertilization than other microorganisms in diverse agricultural soils. Microbiome, 2019, 7(1): 33.
[35]	PII Y, BORRUSO L, BRUSETTI L, CRECCHIO C, CESCO S, MIMMO T. The interaction between iron nutrition, plant species and soil type shapes the rhizosphere microbiome. Plant Physiology and Biochemistry, 2016, 99: 39-48. doi: 10.1016/j.plaphy.2015.12.002 pmid: 26713550
[36]	REN N, WANG Y, YE Y L, ZHAO Y N, HUANG Y F, FU W, CHU X. Effects of continuous nitrogen fertilizer application on the diversity and composition of rhizosphere soil bacteria. Frontiers in Microbiology, 2020, 11: 1948. doi: 10.3389/fmicb.2020.01948 pmid: 32973705
[37]	向君亮, 刘权, 申永瑞, 王佳琦, 张兴梅, 王鹏, 殷奎德. 松嫩草原盐碱土细菌多样性分析. 干旱地区农业研究, 2020, 38(2): 62-68.
	XIANG J L, LIU Q, SHEN Y R, WANG J Q, ZHANG X M, WANG P, YIN K D. Variation of bacterial communities in the saline-alkaline soil of meadow on Songnen Plain. Agricultural Research in the Arid Areas, 2020, 38(2): 62-68. (in Chinese)
[38]	NING L, ZHU C Z, XUE C, CHEN H, DUAN Y, PENG C, GUO S W, SHEN Q. Insight into how organic amendments can shape the soil microbiome in long-term field experiments as revealed by network analysis. Soil Biology and Biochemistry, 2016, 99: 137-149.
[39]	BUCKLEY D H, SCHMIDT T M. Environmental factors influencing the distribution of rRNA from Verrucomicrobia in soil. FEMS Microbiology Ecology, 2001, 35(1): 105-112. doi: 10.1111/j.1574-6941.2001.tb00793.x pmid: 11248395
[40]	KALAM S, BASU A, PODILE A R. Difficult-to-culture bacteria in the rhizosphere: the underexplored signature microbial groups. Pedosphere, 2022, 32(1): 75-89.
[41]	江其朋, 余佳敏, 王金峰, 刘东阳, 龚杰, 江连强, 张淑婷, 余祥文, 李石力, 杨亮, 刘晓姣, 王悦, 王勇, 丁伟. 土壤理化性质驱动烤烟根际细菌群落的组配及其共现性网络互作. 微生物学报, 2023, 63(3): 1168-1184.
	JIANG Q P, YU J M, WANG J F, LIU D Y, GONG J, JIANG L Q, ZHANG S T, YU X W, LI S L, YANG L, LIU X J, WANG Y, WANG Y, DING W. Soil properties affect bacterial community assembly and co-occurrence network in tobacco rhizosphere. Acta Microbiologica Sinica, 2023, 63(3): 1168-1184. (in Chinese)
[42]	DE VRIES F T, GRIFFITHS R I, BAILEY M, CRAIG H, GIRLANDA M, GWEON H S, HALLIN S, KAISERMANN A, KEITH A M, KRETZSCHMAR M, LEMANCEAU P, LUMINI E, MASON K E, OLIVER A, OSTLE N, PROSSER J I, THION C, THOMSON B, BARDGETT R D. Soil bacterial networks are less stable under drought than fungal networks. Nature Communications, 2018, 9: 3033. doi: 10.1038/s41467-018-05516-7 pmid: 30072764
[43]	张梅婷, 刘晋仙, 苏嘉贺, 柴宝峰. 苦草叶表附生和水体浮游细菌群落多样性格局及其影响因素. 环境科学, 2023, 44(1): 252-261.
	ZHANG M T, LIU J X, SU J H, CHAI B F. Diversity patterns and influencing factors of epibiotic in Vallisneria natans and planktonic bacteria communities. Environmental Science, 2023, 44(1): 252-261. (in Chinese)
[44]	刘思博, 殷国梅, 冀超, 丁海君, 薛艳林, 孙林, 阿拉腾布拉格, 高博, 燕茹, 吴昊, 刘永录. 不同浓度氮肥对首年紫花苜蓿产量及其根际土壤真菌群落结构的影响. 中国土壤与肥料, 2023(1): 1-12.
	LIU S B, YIN G M, JI C, DING H J, XUE Y L, SUN L, AALATENG BULAGE, GAO B, YAN R, WU H, LIU Y L. The influence of nitrogen fertilizer application rate on yield of Medicago sativa and the rhizosphere soil fungal diversity. Soil and Fertilizer Sciences in China, 2023(1): 1-12. (in Chinese)
[45]	丁露, 崔立星, 牛营超, 戴小华, 郭青云. 不同氮钾水平对菜豆叶际细菌群落组成的影响. 北方园艺, 2022(4): 76-84.
	DING L, CUI L X, NIU Y C, DAI X H, GUO Q Y. Phyllosphere bacterial community composition on Phaseolus vulgaris Linn under different nitrogen and potassium levels. Northern Horticulture, 2022(4): 76-84. (in Chinese)
[46]	陈懿, 吴春, 李彩斌, 林叶春, 程建中, 潘文杰. 炭基肥对植烟黄壤细菌、真菌群落结构和多样性的影响. 微生物学报, 2020, 60(4): 653-666.
	CHEN Y, WU C, LI C B, LIN Y C, CHENG J Z, PAN W J. Effect of biochar-based fertilizer on bacterial and fungal community composition, diversity in tobacco-planting yellow soil. Acta Microbiologica Sinica, 2020, 60(4): 653-666. (in Chinese)
[47]	LIAO J Y, HU A, ZHAO Z W, LIU X R, JIANG C, ZHANG Z H. Biochar with large specific surface area recruits N₂O-reducing microbes and mitigate N₂O emission. Soil Biology and Biochemistry, 2021, 156: 108212.
[48]	侯雪燕. 土壤pH对硝化作用和氨氧化微生物群落结构的影响[D]. 重庆: 西南大学, 2014.
	HOU X Y. Effects of soil pH on nitrification and community structure of ammonia-oxidizing microorganisms[D]. Chongqing: Southwest University, 2014. (in Chinese)
[49]	孙治强, 张楠, 赵卫星, 高玉红. 氮肥施用量对生菜产量、硝酸盐积累及土壤EC值、pH值的影响. 江西农业学报, 2007, 19(4): 44-45, 48.
	SUN Z Q, ZHANG N, ZHAO W X, GAO Y H. Effects of N application rates on yield, nitrate accumulation of lettuce and EC, pH value of soil. Acta Agriculturae Jiangxi, 2007, 19(4): 44-45, 48. (in Chinese)
[50]	SONG Y, LI X N, YAO S, YANG X L, JIANG X. Correlations between soil metabolomics and bacterial community structures in the pepper rhizosphere under plastic greenhouse cultivation. Science of the Total Environment, 2020, 728: 138439.
[51]	YUAN J, ZHAO J, WEN T, ZHAO M L, LI R, GOOSSENS P, HUANG Q W, BAI Y, VIVANCO J M, KOWALCHUK G A, BERENDSEN R L, SHEN Q R. Root exudates drive the soil-borne legacy of aboveground pathogen infection. Microbiome, 2018, 6(1): 156. doi: 10.1186/s40168-018-0537-x pmid: 30208962
[52]	ISHII S, ASHIDA N, OHNO H, SEGAWA T, YABE S, OTSUKA S, YOKOTA A, SENOO K. Noviherbaspirillum denitrificans sp. nov., a denitrifying bacterium isolated from rice paddy soil and Noviherbaspirillum autotrophicum sp. nov., a denitrifying, facultatively autotrophic bacterium isolated from rice paddy soil and proposal to reclassify Herbaspirillum massiliense as Noviherbaspirillum massiliense comb. nov. International Journal of Systematic and Evolutionary Microbiology, 2017, 67(6): 1841-1848.
[53]	SUNDARARAMAN A, SRINIVASAN S, LEE S S. Noviherbaspirillum humi sp. nov. isolated from soil. Antonie Van Leeuwenhoek, 2016, 109(5): 697-704.
[54]	王敬, 程谊, 蔡祖聪, 张金波. 长期施肥对农田土壤氮素关键转化过程的影响. 土壤学报, 2016, 53(2): 292-304.
	WANG J, CHENG Y, CAI Z C, ZHANG J B. Effects of long-term fertilization on key processes of soil nitrogen cycling in agricultural soil: A review. Acta Pedologica Sinica, 2016, 53(2): 292-304. (in Chinese)
[55]	张博雅, 余珂. 微生物基因数据库在氮循环功能基因注释中的应用. 微生物学通报, 2020, 47(9): 3021-3038.
	ZHANG B Y, YU K. Application of microbial gene databases in the annotation of nitrogen cycle functional genes. Microbiology China, 2020, 47(9): 3021-3038. (in Chinese)
[56]	HU X J, GU H D, LIU J J, WEI D, ZHU P, CUI X A, ZHOU B K, CHEN X L, JIN J, LIU X B, WANG G H. Metagenomics reveals divergent functional profiles of soil carbon and nitrogen cycling under long-term addition of chemical and organic fertilizers in the black soil region. Geoderma, 2022, 418: 115846.
[57]	任栖永. 大豆产量与土壤硝化过程对不同农田管理方式的响应[D]. 杨凌: 西北农林科技大学, 2023.
	REN X Y. Response of soybean yield and soil nitrification process to different farmland management practices[D]. Yangling: Northwest A&F University, 2023. (in Chinese)
[58]	ABDUL R N, PARKS D H, VANWONTERGHEM I, MORRISON M, TYSON G W, HUGENHOLTZ P. A phylogenomic analysis of the bacterial Phylum fibrobacteres. Frontiers in Microbiology, 2016, 6: 1469.

处理 Treatment	放线菌门 Actinobacteria	变形菌门 Proteobacteria	酸杆菌门 Acidobacteria	绿弯菌门 Chloroflexi	芽单胞菌门 Gemmatimonadete	拟杆菌门 Bacteroidetes	疣微菌门 Verrucomicrobia	Patescibacteria	厚壁菌门 Frimicutes	浮霉菌门 Planctomycetes
BN0	36.82	29.79	10.33	8.89	5.31	2.38	1.78	0.86	0.72	0.64
BN168	40.77	29.89	8.45	6.25	4.48	3.39	1.64	1.38	0.53	0.45
BN312	42.12	32.79	6.04	4.73	4.29	4.87	1.18	1.82	0.46	0.28
BN168-BN0	3.95^***	1.21	-1.88^***	-2.64^***	-0.83^***	0.99^***	-0.16	0.52^***	-0.19	-0.19^***
BN312-BN0	5.28^***	2.98^***	-4.29^***	-4.16^***	-1.01^***	2.47^***	-0.62^**	0.96^***	-0.26^*	-0.36^***

处理 Treatment	放线菌门 Actinobacteria	变形菌门 Proteobacteria	酸杆菌门 Acidobacteria	绿弯菌门 Chloroflexi	拟杆菌门 Bacteroidetes	芽单胞菌门 Gemmatimonadete	Patescibacteria	疣微菌门 Verrucomicrobia	厚壁菌门 Frimicutes	浮霉菌门Planctomycetes
SN0	47.99	27.08	7.53	6.88	2.37	3.25	0.94	1.11	1.02	0.33
SN168	39.94	38.70	6.99	3.16	4.01	2.46	1.34	1.07	0.75	0.37
SN312	48.78	33.25	4.96	1.69	4.64	1.90	2.45	0.37	0.67	0.38
SN168-SN0	-8.05^***	11.62^***	-0.54	-3.72^**	1.64^***	-0.79	0.39^***	-0.04	-0.25^**	0.04
SN312-SN0	0.79	6.17^***	-2.57^***	5.19^***	2.23^***	-1.35^***	1.51^***	-0.74^**	-0.33^**	0.05

处理 Treatment	节点数 Node	边数 Edge	平均度 Average degree	平均加权度 Average weighted degree	图密度 Density	模块化 Modularity	正相关 Positive correlation (%)	负相关 Negative correlation (%)
BN0	62	136	3.198	3.152	0.038	0.905	50.74	49.26
BN168	35	123	2.964	2.939	0.036	0.896	61.79	38.21
BN312	58	106	2.437	2.391	0.028	0.894	50.94	49.06
SN0	67	129	2.835	2.777	0.034	0.895	54.26	45.74
SN168	60	117	2.819	2.774	0.032	0.867	51.28	48.72
SN312	60	105	2.386	2.358	0.027	0.867	53.33	46.67