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Article(id=1204800728561857475, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1204800727341310425, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20250449, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1749398400000, receivedDateStr=2025-06-09, revisedDate=null, revisedDateStr=null, acceptedDate=1756051200000, acceptedDateStr=2025-08-25, onlineDate=1765176477803, onlineDateStr=2025-12-08, pubDate=1764777600000, pubDateStr=2025-12-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1765176477803, onlineIssueDateStr=2025-12-08, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1765176477803, creator=13701087609, updateTime=1765176477803, updator=13701087609, issue=Issue{id=1204800727341310425, tenantId=1146029695717560320, journalId=1192105938417971205, year='2025', volume='65', issue='12', pageStart='5191', pageEnd='5649', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1765176477513, creator=13701087609, updateTime=1765176611928, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1204801291189986067, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1204800727341310425, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1204801291189986068, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1204800727341310425, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=5617, endPage=5629, ext={EN=ArticleExt(id=1204800728792544197, articleId=1204800728561857475, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Effects of 3,4-dimethylpyrazole phosphate on ammonia-oxidizing bacteria community in coastal saline-alkaline paddy soil, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

Nitrification inhibitors can affect the biological transformation process of ammonium nitrogen to nitrate nitrogen in soil by inhibiting the activity of ammonia-oxidizing bacteria (AOB). Objective To investigate the effects of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) on the community structure and assembly mechanisms of AOB in coastal saline-alkaline paddy soil. Methods To study the effects of the typical nitrification inhibitor DMPP addition on the diversity, community structure, and community assembly process of AOB in soil under two salinity levels. Pot experiments and high-throughput sequencing were employed to determine the diversity, community structure, and community assembly process of AOB. Results The addition of DMPP increased the alpha diversity of AOB in soil, which reached a significant level in the high-salinity soil. The addition of DMPP significantly changed the community composition of AOB, reducing the relative abundance of taxa with high relative abundance and enriching the taxa with low relative abundance. The decrease in relative abundance of taxa with high relative abundance was the main reason for the inhibition of DMPP on nitrification. Principal coordinates analysis revealed that the community structure of AOB changed significantly after the addition of DMPP, which was more obvious in high-salinity soil. The null model analysis results showed that stochastic processes played a dominant role in the community assembly process of AOB, and the contribution of stochastic processes increased after the addition of DMPP. Canonical correspondence analysis and Mantel’s test indicated that soil pH, electrical conductivity, organic matter, total nitrogen, and alkaline-hydrolyzable nitrogen were the main physicochemical factors influencing changes in AOB community structure. Conclusion DMPP exerted significant impacts on AOB communities in coastal saline-alkaline paddy soils across varying salinity levels, with its inhibitory effects varying substantially with soil salinity.

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*E-mail:
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硝化抑制剂可通过抑制氨氧化细菌(ammonia-oxidizing bacteria, AOB)的活性影响土壤中铵态氮转化为硝态氮的生物转化过程。 目的 研究硝化抑制剂3,4-二甲基吡唑磷酸盐(3,4-dimethylpyrazole phosphate, DMPP)对滨海盐碱水稻土中AOB群落结构及其构建机制的影响。 方法 以典型的硝化抑制剂DMPP为材料，基于盆栽试验和高通量测序技术研究在2种盐度条件下添加DMPP对土壤AOB多样性、群落结构和群落组装过程的影响。 结果 添加DMPP提高了土壤AOB群落的α多样性，且在高盐土壤中达到了显著水平；添加DMPP显著改变了AOB的群落组成，降低了高相对丰度类群的相对丰度，富集低相对丰度类群，且高相对丰度类群AOB相对丰度的降低是DMPP抑制硝化作用过程的主要原因；主坐标分析发现，添加DMPP后AOB的群落结构发生明显变化，且高盐土壤中的变化大于低盐土壤；零模型分析结果显示，随机性过程在AOB群落组装过程中起主导作用，且添加DMPP后随机性过程的贡献更大；典范对应分析和曼特尔检验表明，土壤酸碱度、电导率、有机质、总氮和碱解氮是影响AOB群落结构变化的主要理化因子。 结论 添加DMPP对不同盐分含量的滨海盐碱水稻土中AOB群落具有显著影响，且DMPP在不同盐度土壤中的抑制效果不同。

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Bioresource Technology, 2016, 216: 714-721., articleTitle=Response of performance and ammonia oxidizing bacteria community to high salinity stress in membrane bioreactor with elevated ammonia loading, refAbstract=null), Reference(id=1217784604501660286, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, doi=null, pmid=null, pmcid=null, year=2021, volume=9, issue=9, pageStart=1929, pageEnd=null, url=null, language=null, rfNumber=[31], rfOrder=35, authorNames=LIU NN, HU HF, MA WH, DENG Y, WANG QG, LUO A, MENG JH, FENG XJ, WANG ZH, journalName=Microorganisms, refType=null, unstructuredReference=LIU NN, HU HF, MA WH, DENG Y, WANG QG, LUO A, MENG JH, FENG XJ, WANG ZH. Relative importance of deterministic and stochastic processes on soil microbial community assembly in temperate grasslands[J]. Microorganisms, 2021, 9(9): 1929., articleTitle=Relative importance of deterministic and stochastic processes on soil microbial community assembly in temperate grasslands, refAbstract=null)], funds=[Fund(id=1217784599313305913, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, awardId=32471725, language=EN, fundingSource=National Natural Science Foundation of China(32471725), fundOrder=null, country=null), Fund(id=1217784599422357827, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, awardId=32471725, language=CN, fundingSource=国家自然科学基金(32471725), fundOrder=null, country=null), Fund(id=1217784599560769870, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, awardId=2022Z169, language=EN, fundingSource=Major Special Projects of “Science and Technology Innovation 2025” in Ningbo(2022Z169), fundOrder=null, country=null), Fund(id=1217784599674016084, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, awardId=2022Z169, language=CN, fundingSource=宁波市“科技创新2025”重大专项(2022Z169), fundOrder=null, country=null), Fund(id=1217784599808233824, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, awardId=2023Z114, language=EN, fundingSource=Major Special Projects of “Science and Technology Innovation 2025” in Ningbo(2023Z114), fundOrder=null, country=null), Fund(id=1217784599925674344, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, awardId=2023Z114, language=CN, fundingSource=宁波市“科技创新2025”重大专项(2023Z114), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1217784592837301076, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, xref=1., ext=[AuthorCompanyExt(id=1217784592866661207, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, companyId=1217784592837301076, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture and Rural Affairs, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, China), AuthorCompanyExt(id=1217784592896021337, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, companyId=1217784592837301076, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.南京农业大学 生命科学学院，农业农村部农业环境微生物重点实验室，江苏 南京)]), AuthorCompany(id=1217784593051210591, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, xref=2., ext=[AuthorCompanyExt(id=1217784593063793505, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, companyId=1217784593051210591, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.Ningbo Key Laboratory of Quality and Safety Detection and Control of Characteristic Agricultural Products, Ningbo Academy of Agricultural Sciences, Ningbo, Zhejiang, China), AuthorCompanyExt(id=1217784593105736549, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, companyId=1217784593051210591, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.宁波市农业科学研究院，宁波市特色农产品质量安全检测与控制重点实验室，浙江 宁波)])], figs=[ArticleFig(id=1217784597170016410, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=EN, label=Figure 1, caption=Analysis of soil AOB community diversity. A: Alpha diversity analysis of soil AOB communities; B: Principal component analysis of soil AOB communities. Different lowercase letters indicate significant differences between treatments (P<0.05)., figureFileSmall=wrApnZ2tK5eMIQS2joHkCg==, figureFileBig=5h2yHs8+sEAV/YYLw0xjPQ==, tableContent=null), ArticleFig(id=1217784597316817061, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=CN, label=图1, caption=土壤AOB群落多样性分析。A：土壤AOB群落的α多样性分析；B：土壤AOB群落的主坐标分析。不同小写字母表示处理间的差异显著性(P<0.05)。, figureFileSmall=wrApnZ2tK5eMIQS2joHkCg==, figureFileBig=5h2yHs8+sEAV/YYLw0xjPQ==, tableContent=null), ArticleFig(id=1217784597505560755, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=EN, label=Figure 2, caption=Volcano plot of OTU differences in AOB community under different treatments. The x-axis log10 baseMean represents the average abundance of OTUs in samples. OTUs with log10 baseMean>2 are defined as high relative abundance taxa, and OTUs with log10 baseMean<2 are defined as low relative abundance taxa. The y-axis log2 fold change indicates the degree of difference in OTUs abundance changes. log2 fold change>1 indicates increased abundance, represented in yellow, while log2 fold change<-1 indicates decreased abundance, represented in blue., figureFileSmall=zBXaxRmZlXYeB62fbQ3zew==, figureFileBig=2QFRshSmmVbxSEChEZa3Dg==, tableContent=null), ArticleFig(id=1217784597639778490, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=CN, label=图2, caption=不同处理AOB群落OTU差异火山图。横坐标log10 baseMean表示OTUs在样本中的平均丰度，将log10 baseMean>2的OTUs定义为高相对丰度类群，将log10 baseMean<2的OTUs定义为低相对丰度类群；纵坐标log2 fold chang表示OTUs丰度变化的差异程度，log2 fold chang>1表示丰度升高，用黄色表示，log2 fold chang<-1表示丰度降低，用蓝色表示。, figureFileSmall=zBXaxRmZlXYeB62fbQ3zew==, figureFileBig=2QFRshSmmVbxSEChEZa3Dg==, tableContent=null), ArticleFig(id=1217784597744636097, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=EN, label=Figure 3, caption=Phylogenetic tree of AOB amoA gene sequences (A) and stacked bar plot showing the relative abundance of AOB phylogenetic groups (B). In the phylogenetic tree, OTUs represent sequences with relative abundance >1%, while “Others” in the bar plot denotes OTUs with relative abundance <1%., figureFileSmall=s/Ng55ReloffjmpVQlsc1Q==, figureFileBig=q4d04++UcHK8YA85QkD12A==, tableContent=null), ArticleFig(id=1217784597857882315, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=CN, label=图3, caption=AOB的 amoA 基因序列系统发育树(A)和AOB系统发育类群相对丰度堆叠柱状图(B)。系统发育树中的OTUs为相对丰度>1%的代表性序列，柱状图中的Others表示相对丰度<1%的其他OTUs。, figureFileSmall=s/Ng55ReloffjmpVQlsc1Q==, figureFileBig=q4d04++UcHK8YA85QkD12A==, tableContent=null), ArticleFig(id=1217784597992100052, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=EN, label=Figure 4, caption=Analysis of the soil AOB community assembly process. A: βNTI values of AOB communities in different treatments; B: Percentage of deterministic and stochastic assembly processes of AOB communities in different treatments., figureFileSmall=b6zPbF7HrHj0QgRh7/bfdw==, figureFileBig=33rJ4CuCw1E1h8qj6sHTbQ==, tableContent=null), ArticleFig(id=1217784598226981094, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=CN, label=图4, caption=土壤AOB群落组装过程分析。A：不同处理AOB群落的βNTI值；B：不同处理AOB群落确定性和随机性组装过程的百分比。, figureFileSmall=b6zPbF7HrHj0QgRh7/bfdw==, figureFileBig=33rJ4CuCw1E1h8qj6sHTbQ==, tableContent=null), ArticleFig(id=1217784598357004533, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=EN, label=Figure 5, caption=Correlation analysis between the soil AOB community and environmental factors. A: Canonical correspondence analysis of soil AOB community structure; B: Mantel’s test, cluster analysis, and Spearman’s correlation heatmap between soil AOB community structure and physicochemical factors. The asterisks indicate significant correlations between the two, with *P<0.05, **P<0.01, and ***P<0.001., figureFileSmall=9s39qsswrLyL68PyPIgdlw==, figureFileBig=Hw8h8CXzf6x9zuhDaUG/Wg==, tableContent=null), ArticleFig(id=1217784598487027971, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=CN, label=图5, caption=土壤AOB群落与环境因子相关性分析。A：土壤AOB群落结构的典范对应分析；B：土壤AOB群落结构与理化因子的Mantel检验、聚类分析及Spearman相关性热图。, figureFileSmall=9s39qsswrLyL68PyPIgdlw==, figureFileBig=Hw8h8CXzf6x9zuhDaUG/Wg==, tableContent=null), ArticleFig(id=1217784598608662794, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=EN, label=Table 1, caption=

The soil physical and chemical properties of different treatments

, figureFileSmall=null, figureFileBig=null, tableContent=

处理

Treatment

酸碱度

Potential

of hydrogen

电导率

Electrical

conductivity (μS/cm)

有机质

Soil organic matter

(g/kg)

有效磷

Available

phosphorus (mg/kg)

速效钾

Available

potassium

(mg/kg)

全氮

Total

nitrogen (g/kg)

碱解氮

Alkaline hydrolyzable nitrogen (mg/kg)

HCK8.83±0.11a535.67±43.10a8.92±0.14b66.32±7.84a407.00±50.09a0.79±0.08b110.13±1.76c
HDMPP8.79±0.09a530.67±38.08a8.57±0.21b45.19±2.91bc382.67±74.66a0.79±0.04b96.03±7.15c
LCK8.60±0.06b287.33±25.50b15.95±1.20a46.75±7.24b438.67±20.55a1.70±0.08a152.17±2.97b
LDMPP8.38±0.12c281.33±18.77b15.13±0.38a34.71±2.80c463.33±26.08a1.54±0.16a181.13±16.57a
), ArticleFig(id=1217784598700937488, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=CN, label=表1, caption=

不同处理的土壤理化性质

, figureFileSmall=null, figureFileBig=null, tableContent=

处理

Treatment

酸碱度

Potential

of hydrogen

电导率

Electrical

conductivity (μS/cm)

有机质

Soil organic matter

(g/kg)

有效磷

Available

phosphorus (mg/kg)

速效钾

Available

potassium

(mg/kg)

全氮

Total

nitrogen (g/kg)

碱解氮

Alkaline hydrolyzable nitrogen (mg/kg)

HCK8.83±0.11a535.67±43.10a8.92±0.14b66.32±7.84a407.00±50.09a0.79±0.08b110.13±1.76c
HDMPP8.79±0.09a530.67±38.08a8.57±0.21b45.19±2.91bc382.67±74.66a0.79±0.04b96.03±7.15c
LCK8.60±0.06b287.33±25.50b15.95±1.20a46.75±7.24b438.67±20.55a1.70±0.08a152.17±2.97b
LDMPP8.38±0.12c281.33±18.77b15.13±0.38a34.71±2.80c463.33±26.08a1.54±0.16a181.13±16.57a
), ArticleFig(id=1217784598789017884, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=EN, label=Table 2, caption=

Soil ammonium nitrogen and nitrate nitrogen contents under different treatments across rice growth stages

, figureFileSmall=null, figureFileBig=null, tableContent=

处理

Treatment

分蘖期Tillering stage抽穗期Heading stage成熟期Ripening stage
NO3--N (mg/kg)NH4+-N (mg/kg)NO3--N (mg/kg)NH4+-N (mg/kg)NO3--N (mg/kg)NH4+-N (mg/kg)
HCK4.14±0.40a113.22±4.61d0.070±0.026a12.36±7.66d3.65±1.02a0.81±0.26c
HDMPP0.24±0.11c123.89±7.38c0.085±0.010a45.64±11.24c0.70±0.03b4.93±0.82b
LCK1.34±0.20b135.36±6.46b0.047±0.013a66.26±9.71b3.05±0.76a0.34±0.19c
LDMPP0.38±0.11c221.69±15.69a0.085±0.015a118.45±7.66a0.43±0.04b15.96±0.87a
), ArticleFig(id=1217784598990344488, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1204800728561857475, language=CN, label=表2, caption=

不同处理下水稻各生育期土壤硝态氮与铵态氮含量

, figureFileSmall=null, figureFileBig=null, tableContent=

处理

Treatment

分蘖期Tillering stage抽穗期Heading stage成熟期Ripening stage
NO3--N (mg/kg)NH4+-N (mg/kg)NO3--N (mg/kg)NH4+-N (mg/kg)NO3--N (mg/kg)NH4+-N (mg/kg)
HCK4.14±0.40a113.22±4.61d0.070±0.026a12.36±7.66d3.65±1.02a0.81±0.26c
HDMPP0.24±0.11c123.89±7.38c0.085±0.010a45.64±11.24c0.70±0.03b4.93±0.82b
LCK1.34±0.20b135.36±6.46b0.047±0.013a66.26±9.71b3.05±0.76a0.34±0.19c
LDMPP0.38±0.11c221.69±15.69a0.085±0.015a118.45±7.66a0.43±0.04b15.96±0.87a
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3,4-二甲基吡唑磷酸盐对滨海盐碱水稻土氨氧化细菌群落的影响
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周家庆 1 , 徐敏 1 , 汪峰 2 , 刘浩 1 , 朱诗君 2 , 曹慧 1, *
微生物学报 | 研究报告 2025,65(12): 5617-5629
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微生物学报 | 研究报告 2025, 65(12): 5617-5629
3,4-二甲基吡唑磷酸盐对滨海盐碱水稻土氨氧化细菌群落的影响
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周家庆1, 徐敏1, 汪峰2, 刘浩1, 朱诗君2, 曹慧1, *
作者信息
  • 1.南京农业大学 生命科学学院，农业农村部农业环境微生物重点实验室，江苏 南京
  • 2.宁波市农业科学研究院，宁波市特色农产品质量安全检测与控制重点实验室，浙江 宁波
Effects of 3,4-dimethylpyrazole phosphate on ammonia-oxidizing bacteria community in coastal saline-alkaline paddy soil
Jiaqing ZHOU1, Min XU1, Feng WANG2, Hao LIU1, Shijun ZHU2, Hui CAO1, *
Affiliations
  • 1.Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture and Rural Affairs, College of Life Sciences, Nanjing Agricultural University, Nanjing, Jiangsu, China
  • 2.Ningbo Key Laboratory of Quality and Safety Detection and Control of Characteristic Agricultural Products, Ningbo Academy of Agricultural Sciences, Ningbo, Zhejiang, China
出版时间: 2025-12-04 doi: 10.13343/j.cnki.wsxb.20250449
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摘要
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硝化抑制剂可通过抑制氨氧化细菌(ammonia-oxidizing bacteria, AOB)的活性影响土壤中铵态氮转化为硝态氮的生物转化过程。 目的 研究硝化抑制剂3,4-二甲基吡唑磷酸盐(3,4-dimethylpyrazole phosphate, DMPP)对滨海盐碱水稻土中AOB群落结构及其构建机制的影响。 方法 以典型的硝化抑制剂DMPP为材料，基于盆栽试验和高通量测序技术研究在2种盐度条件下添加DMPP对土壤AOB多样性、群落结构和群落组装过程的影响。 结果 添加DMPP提高了土壤AOB群落的α多样性，且在高盐土壤中达到了显著水平；添加DMPP显著改变了AOB的群落组成，降低了高相对丰度类群的相对丰度，富集低相对丰度类群，且高相对丰度类群AOB相对丰度的降低是DMPP抑制硝化作用过程的主要原因；主坐标分析发现，添加DMPP后AOB的群落结构发生明显变化，且高盐土壤中的变化大于低盐土壤；零模型分析结果显示，随机性过程在AOB群落组装过程中起主导作用，且添加DMPP后随机性过程的贡献更大；典范对应分析和曼特尔检验表明，土壤酸碱度、电导率、有机质、总氮和碱解氮是影响AOB群落结构变化的主要理化因子。 结论 添加DMPP对不同盐分含量的滨海盐碱水稻土中AOB群落具有显著影响，且DMPP在不同盐度土壤中的抑制效果不同。

3,4-二甲基吡唑磷酸盐  /  氨氧化细菌  /  群落多样性  /  群落组成  /  群落组装

Nitrification inhibitors can affect the biological transformation process of ammonium nitrogen to nitrate nitrogen in soil by inhibiting the activity of ammonia-oxidizing bacteria (AOB). Objective To investigate the effects of the nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) on the community structure and assembly mechanisms of AOB in coastal saline-alkaline paddy soil. Methods To study the effects of the typical nitrification inhibitor DMPP addition on the diversity, community structure, and community assembly process of AOB in soil under two salinity levels. Pot experiments and high-throughput sequencing were employed to determine the diversity, community structure, and community assembly process of AOB. Results The addition of DMPP increased the alpha diversity of AOB in soil, which reached a significant level in the high-salinity soil. The addition of DMPP significantly changed the community composition of AOB, reducing the relative abundance of taxa with high relative abundance and enriching the taxa with low relative abundance. The decrease in relative abundance of taxa with high relative abundance was the main reason for the inhibition of DMPP on nitrification. Principal coordinates analysis revealed that the community structure of AOB changed significantly after the addition of DMPP, which was more obvious in high-salinity soil. The null model analysis results showed that stochastic processes played a dominant role in the community assembly process of AOB, and the contribution of stochastic processes increased after the addition of DMPP. Canonical correspondence analysis and Mantel’s test indicated that soil pH, electrical conductivity, organic matter, total nitrogen, and alkaline-hydrolyzable nitrogen were the main physicochemical factors influencing changes in AOB community structure. Conclusion DMPP exerted significant impacts on AOB communities in coastal saline-alkaline paddy soils across varying salinity levels, with its inhibitory effects varying substantially with soil salinity.

3,4-dimethylpyrazole phosphate (DMPP)  /  ammonia-oxidizing bacteria (AOB)  /  community diversity  /  community composition  /  community assembly
周家庆, 徐敏, 汪峰, 刘浩, 朱诗君, 曹慧. 3,4-二甲基吡唑磷酸盐对滨海盐碱水稻土氨氧化细菌群落的影响. 微生物学报, 2025 , 65 (12) : 5617 -5629 . DOI: 10.13343/j.cnki.wsxb.20250449
Jiaqing ZHOU, Min XU, Feng WANG, Hao LIU, Shijun ZHU, Hui CAO. Effects of 3,4-dimethylpyrazole phosphate on ammonia-oxidizing bacteria community in coastal saline-alkaline paddy soil[J]. Acta Microbiologica Sinica, 2025 , 65 (12) : 5617 -5629 . DOI: 10.13343/j.cnki.wsxb.20250449
正文
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硝化作用是由微生物介导的、将铵(NH4+)氧化为硝酸盐(NO3-)的过程，是氮循环的重要环节，对土壤微生物和植物氮基质(NH4+-N或NO3--N)的可用性具有重要影响。硝化过程产生的硝酸盐(NO3-)淋失会导致农业系统中的氮损失[1]，还会引发水体富营养化和饮用水污染。抑制土壤中的硝化作用是最大限度减少农业系统氮损失的重要策略，将硝化抑制剂与氮肥联合施用是减轻硝化作用的有效方法[2]。DMPP是一种新型硝化抑制剂，在抑制土壤硝化作用、提高氮肥利用率、增加作物产量和提升作物品质方面效果显著，且其施用剂量比其他硝化抑制剂小，具有高效、无毒、环保和性价比高的特点[3]。
氨氧化反应由AOB、氨氧化古细菌(ammonia-oxidizing archaea, AOA)和全程氨氧化菌(complete ammonia oxidizers, Comammox)中的amoA基因编码的氨单加氧酶(ammonia monooxygenase, AMO)催化，DMPP通过改变AMO的活性位点并形成多肽，使AMO失活进而影响氨氧化反应[4]。一般而言，AOB在中性和碱性土壤的硝化过程中占主导地位，AOA和Comammox更倾向于酸性土壤环境[5]，而绝大多数盐碱地土壤为碱性土壤。施用含有硝化抑制剂的肥料时土壤中AOB的丰度会受到抑制，而AOA、Comammox和非目标微生物不会受到负面影响[6]。然而，以往研究通常关注硝化抑制剂对AOB丰度的影响，对AOB多样性、群落组成及组装过程的系统性探讨仍相对不足[7]。由于微生物群落结构决定微生物功能，阐明AOB群落结构变化有助于更好地理解土壤硝化作用的过程机制。此外，尽管DMPP已被证明能有效抑制土壤硝化作用，但其功效在不同土壤类型间存在显著差异，温度、湿度、土壤酸碱度和土壤有机质[8-9]对DMPP抑制效果的影响已得到广泛研究，而有关土壤盐度对DMPP功效影响的研究较少，有研究指出土壤电导率是影响DMPP功效的主导因素[10]，但该研究针对的是中性和微酸性土壤，目前仍缺乏关于盐碱土中盐度如何影响DMPP抑制效果的系统研究。我国盐碱地面积大，开发利用潜力广阔，盐碱地已成为至关重要的后备耕地资源和“潜在粮仓”[11]。因此，开展土壤盐度和DMPP作用效果之间关系的研究具有重要的理论和实践意义。
本研究采用盆栽试验和高通量测序的方法，通过向2种盐度的土壤施用DMPP探究其对AOB多样性、群落组成和组装过程的影响，以及土壤盐度对DMPP抑制效果的影响，揭示AOB群落对DMPP施用的响应机制，以期为合理施用DMPP降低农田土壤氮损失提供理论依据。
1 材料与方法
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1.1 供试土样与实验设计
供试土样采自慈溪市杭州湾滨海农区(30°20′N, 121°30′E)。该地区属亚热带季风气候，雨量充沛，年平均降水量为1 343.1 mm，年平均日照时数为1 861.5 h，年平均温度为16.9 ℃。选择该地区典型稻麦轮作系统下的农田，在水稻种植前分别于序号为11和4的农田中采集代表高盐和低盐的表层土壤(0-20 cm)，土壤类型为海相沉积物发育而成的盐化潮土。盆栽试验设置在宁波市农业科学研究院横溪试验基地的玻璃温室。供试土壤经自然风干、去除杂质、磨碎后过5 mm筛，然后分别装入塑料花盆中，每盆装土8 kg。盆栽前将土泡水1周。每盆施入10 g尿素和6 g KH2PO4，尿素在基肥、分蘖肥和穗肥阶段分段施入，比例为4:3:3；KH2PO4作为基肥和穗肥分2次等量施入。基肥(4 g尿素+3 g KH2PO4+0.2 g DMPP)于2016年7月14日施用，分蘖肥(3 g尿素+0.2 g DMPP)于2016年7月28日施用，穗肥(3 g尿素+3 g KH2PO4+0.2 g DMPP)于2016年8月20日施用。
试验设高盐(H)和低盐(L) 2种盐分处理，盐浓度通过土壤电导率表征。每种盐分处理下进一步设置2类DMPP处理：添加DMPP (HDMPP和LDMPP)与不添加DMPP (HCK和LCK)，共4种处理，每种处理均设置3个生物学重复，以保证结果的可靠性。水稻品种为‘甬优12’，每盆栽培6株水稻苗，除施肥外，水稻的除草、喷洒农药等病虫害管理均参照当地的常规管理模式进行。
1.2 土壤样本采集
在水稻分蘖期、抽穗期分别使用无菌刀片在每盆土壤中随机采集3份土样用于测定土壤中铵态氮和硝态氮的含量。在水稻成熟期收获后采集盆栽土样，去除作物残体等杂质后充分混匀，随后将土壤分成2部分：一部分暂存于-80 ℃冰箱中用于后续DNA提取和高通量测序；另一部分土样置于室内室温下自然风干，研磨后分别过孔径为1.00、0.25和0.15 mm的标准筛，用于土壤理化性质的测定。
1.3 土壤理化性质测定
土壤基本理化性质的测定均参考鲍士旦[12]的方法。土壤酸碱度(potential of hydrogen, pH)使用pH计进行测定(水土比为2.5:1)；土壤电导率(electrical conductivity, EC)使用电导率仪进行测定(水土比为5:1)；土壤有机质(soil organic matter, SOM)使用重铬酸钾容量法进行测定；土壤总氮(total nitrogen, TN)使用凯氏定氮法进行测定；土壤碱解氮(alkali-hydrolyzable nitrogen, AN)使用碱解扩散法进行测定；土壤有效磷(available phosphorus, AP)使用碳酸氢钠浸提-钼锑钪比色法进行测定；土壤速效钾(available potassium, AK)使用乙酸铵浸提——火焰光度法进行测定；土壤铵态氮(NH4+-N)和硝态氮(NO3--N)用2 mol/L KCl溶液浸提，用连续流动分析仪进行测定。
1.4 土壤DNA提取和高通量测序
按照E.Z.N.A.® Soil DNA Kit (Omega Bio-tek公司)的说明提取土壤总DNA，用1%琼脂糖凝胶电泳检测提取的DNA，并用NanoQuant (Tecan公司)检测DNA的浓度和纯度。选用引物amoA-1F (5′-GGGGTTTCTACTGGTGGT-3′)和amoA-2R (5′-CCCCTCKGSAAAGCCTTCTTC-3′)[13]扩增AOB的amoA基因。PCR扩增体系(50 μL)：Ex Taq (10×)缓冲液5.0 μL，dNTPs 4.0 μL，上、下游引物(10 µmol/L)各1 µL，Ex Taq DNA聚合酶(5 U/μL) 0.5 μL，DNA模板2.0 μL，加ddH2O至50 μL。PCR扩增条件：95 ℃预变性3 min；94 ℃变性45 s，55 ℃退火30 s，72 ℃延伸45 s，循环35次。使用NanoDrop ND-1000 UV-Vis分光光度计检测PCR产物的数量和质量。扩增成功后，将PCR产物送至南京擎科生物科技有限公司进行MiSeq平台高通量测序分析。
使用QIIME流程(v1.9.0)[14]对原始测序序列进行处理。首先，通过Cutadapt (v1.9.1)进行质量过滤，剔除低质量测序reads，以获取高质量的序列；采用UCHIME方法对嵌合序列进行检测和删除。测序后的序列经过优化和去杂后，12个样品的总测序量为309 636条，每个样本的序列数从18 140到30 725不等，平均为25 803条。然后，利用QIIME中的UCLUST功能对序列进行聚类，设定相似性阈值为≥97%[15]，生成可操作分类单元(operational taxonomic unit, OTU)。将相对丰度大于1%的OTU的代表性序列与NCBI数据库中的参考序列一起构建系统发育树，判断其分类地位。核酸序列原始数据存储在国家微生物科学数据中心(http://nmdc.cn)，编号为NMDCN0008L4R。
1.5 数据分析
使用SPSS 25.0进行单因素方差分析以确定各处理之间理化性质的差异显著性。利用R中的“vegan”包计算Chao1、ACE丰富度指数、Shannon多样性指数和Pielou均匀度指数，采用Tukey HSD检验进行方差分析。基于Bray-Curtis距离进行主坐标分析(principal coordinates analysis, PCoA)和非参数多元方差分析(PERMANOVA)。利用MEGA 11软件中的邻接法构建系统发育树。利用R中的“vegan”包进行典范对应分析(canonical correspondence analysis, CCA)和Mantel检验分析环境因子与AOB群落结构之间的相关性；利用R中的“picante”包进行零模型(null model)分析，计算βNTI (β nearest taxon index)和基于Bray-Curtis的RCbray (Raup-Crick)指数，根据两者的数值量化异质选择、同质选择、扩散限制、同质扩散以及未主导过程对AOB群落组装的贡献。
2 结果与分析
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2.1 土壤理化性质
从表1可以看出，高盐土壤与低盐土壤的理化性质存在显著差异。高盐土壤的酸碱度、电导率和有效磷含量显著高于低盐土壤，而有机质、全氮和碱解氮含量显著低于低盐土壤(P<0.05)。添加DMPP显著降低了水稻分蘖期和成熟期2种土壤的硝态氮含量，显著增加了水稻各个生育期时2种土壤的铵态氮含量；硝态氮含量随水稻生育期的推移呈U形分布，而铵态氮含量随生育期下降(表2)。
2.2 DMPP对土壤AOB群落α和β多样性的影响
不同处理土壤AOB群落的α多样性如图1A所示，其中Chao1和ACE指数表示AOB丰富度，Shannon和Pielou指数分别表示多样性和均匀度。高盐土壤的AOB丰富度、多样性和均匀度高于低盐土壤，但差异不显著。在高盐土壤环境条件下，HDMPP处理的土壤AOB丰富度、多样性和均匀度显著提高；而在低盐土壤环境条件下，LDMPP处理的土壤AOB丰富度、多样性和均匀度有一定程度的提高，但差异不显著。
主坐标分析结果如图1B所示，主轴1 (PC1)和主轴2 (PC2)分别解释了AOB群落变异的51.22%和13.31%。从PC1角度分析，高盐土壤与低盐土壤的AOB群落结构均有显著差异(P<0.05)，但DMPP处理与CK处理的AOB群落结构相似，无显著区分。从PC2角度分析，HCK处理与HDMPP处理的AOB群落结构有显著差异(P<0.05)，其余处理的AOB群落结构无显著区分。
2.3 DMPP对土壤AOB群落组成的影响
为了阐明DMPP对AOB群落组成的影响，分析了不同处理间AOB在OTU水平上的丰度差异变化，结果如图2所示。与HCK处理相比，HDMPP处理中有65个OTUs丰度降低，130个OTUs丰度升高；与LCK处理相比，LDMPP处理中有25个OTUs丰度降低，45个OTUs丰度升高，且降低的OTUs多为高相对丰度OTUs，升高的OTUs多为低相对丰度OTUs。
使用优势OTUs (相对丰度>1%)构建AOB的系统发育树。AOB系统发育树包含5个分支：亚硝化螺菌属(Nitrosospira Cluster 3a)、Nitrosospira Cluster 3b以及亚硝化单胞菌属(Nitrosomonasoligotropha)、N. eutropha和N. communis (图3A)。与HCK处理相比，HDMPP处理中N. eutropha相对丰度显著降低；与LCK处理相比，LDMPP处理中N. communis相对丰度降低(图3B)。这些结果表明DMPP处理与CK处理之间的AOB群落组成存在明显的差异，尤其是一些高相对丰度类群在2种处理之间差异显著。
2.4 不同处理土壤AOB群落组装机制
采用βNTI表征AOB群落的组装过程，并量化了不同组装过程的相对贡献(图4)。在4种处理中，随机性过程在AOB群落组装中起主导作用。与CK处理相比，DMPP处理中AOB群落的随机性过程比例上升，在HDMPP处理中达到94.44%，在LDMPP处理中达到86.11%。随机性过程比例的上升主要归因于扩散限制比例的增加。与CK处理相比，DMPP处理中AOB群落的确定性过程比例下降，在HDMPP处理中仅为5.56%，而在LDMPP处理中仅为13.89%。确定性过程比例的下降是由同质选择比例降低导致的。
2.5 土壤AOB群落与环境因子相关性分析
土壤AOB与环境因子的CCA分析结果见图5A。前2个排序轴对AOB群落变异的解释值分别为19.18%和14.78%，共解释了33.96%的群落变异信息。其中，pH (P=0.003)、EC (P=0.002)、SOM (P=0.002)、TN (P=0.004)、AN (P=0.001)均达到显著水平，表明它们是影响土壤AOB群落组成的主要环境因子。Mantel检验结果表明，HCK处理土壤AOB群落与pH、AP、NO3--N呈显著相关；HDMPP处理土壤AOB群落与SOM和AN呈显著相关；LCK处理土壤AOB群落与SOM、TN、NO3--N、NH4+-N呈显著相关；LDMPP处理土壤AOB群落与pH、EC、AP、NO3--N、NH4+-N和铵硝比显著相关。
3 讨论
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3.1 DMPP对土壤AOB多样性和群落组成的影响
本研究中，DMPP对土壤AOB群落多样性和组成产生了显著影响。尽管已有研究普遍认为DMPP会影响AOB的丰度和活性，但关于其多样性和群落组成方面的研究结果较少。Yin等[16]研究表明，虽然DMPP主要通过抑制AOB的丰度来降低土壤铵态氮的硝化速率，但DMPP对AOB群落的多样性无显著影响，这与本研究的结果相反。在本研究中，DMPP对AOB的群落组成和多样性存在显著影响。与CK处理相比，DMPP处理中上调的OTU数量为下调的OTU数量的2倍左右，且下调的OTU多为相对丰度较高的OTU，富集的OTU多为相对丰度较低的OTU，可见DMPP会抑制AOB高相对丰度类群的生长，高相对丰度类群相对丰度减少导致生态位空缺，从而有利于低相对丰度类群的生长。同时，DMPP显著提高了高盐土壤中AOB群落的多样性，低盐土壤中AOB群落的多样性也有一定程度的提高，DMPP通过抑制优势AOB的氨氧化活性导致土壤NH4+积累，高NH4+环境进一步抑制了少数高相对丰度类群AOB的生长，使其相对丰度降低，促进低相对丰度类群AOB的生态位扩张，最终表现为群落多样性的升高。可以合理推测，高相对丰度类群AOB相对丰度降低引起土壤AOB数量减少，这是DMPP抑制硝化作用过程的主要原因。AOB多样性和群落组成在高盐与低盐土壤间表现出的差异则可能归因于土壤理化性质对DMPP有效性和持久性的影响。从不同处理水稻各生育期时土壤硝态氮的含量可以看出，DMPP在高盐土壤中的硝化抑制率高于低盐土壤。高盐土壤EC值较高，土壤溶液中可溶性离子的浓度较高，可能通过静电排斥减少了DMPP分子的分子间相互作用，从而提高了DMPP的有效性[17]；另一方面，低盐土壤有机质含量较高，DMPP易被土壤中的有机质吸附，从而影响DMPP的持久性和有效性[10]。DMPP作用效果增强后对AOB高相对丰度类群生长的抑制作用也随之增强，更有利于低相对丰度类群的生长，进一步提高AOB群落多样性，从而导致多样性的升高在高盐与低盐土壤间表现出差异。
3.2 DMPP增加随机性过程在AOB群落组装过程中的贡献
群落组装在塑造微生物群落的多样性和组成方面至关重要。确定性过程和随机性过程共同作用于微生物群落的组装过程，但这些过程在驱动微生物群落组装中的相对贡献仍存在争议[18]。有关功能微生物群落组装机制的研究较少。Feng等[19]研究了水稻土壤中全程氨氧化菌群落的组装机制，表明随机性过程主导了群落的组装，这与本研究结果一致。研究表明确定性过程在土壤养分条件低的土壤中占主导地位，而随机性过程在土壤养分条件高的土壤中占主导地位[20]。本研究受试土壤是养分丰富的农田土壤，丰富的养分可通过促进微生物生长和定殖来削弱选择压力，从而增强了随机性过程在微生物群落组装中的重要性。与CK处理相比，DMPP处理的AOB群落中随机性过程的比例升高。研究表明种间资源竞争压力的削弱会增强随机性过程在微生物群落组装过程中的重要性[21]。DMPP对少数AOB高相对丰度类群生长的抑制在一定程度上减缓了其他AOB竞争资源的压力。研究表明随机性过程能够使微生物群落产生更多样化的生态功能，有助于维持微生物群落的稳定性和持续性[22]。同时，由随机性过程主导的微生物群落能够对外界环境变化所带来的干扰提供一定的缓冲作用，因此对系统的稳定性有一定的正反馈调节效果[23]。DMPP处理中AOB群落增加的随机性过程可提供更高的微生物多样性，能够部分缓解DMPP对AOB群落造成的干扰。Xun等[24]的研究表明，在微生物群落组装过程中随机性过程占比越高微生物的多样性就越高；而确定性过程占比越高微生物的多样性就越低，这进一步佐证了本研究的结果。
3.3 土壤AOB群落与环境因子之间的关系
土壤pH、有机质含量、养分含量等理化性质与微生物群落的多样性和组成密切相关[25]。本研究发现，土壤有机质含量、电导率和pH是影响盐碱地土壤AOB群落结构的主要环境因子。土壤有机质作为土壤理化性质的重要指标之一，是土壤养分循环转化的核心，其含量的变化对土壤肥力和土壤微生物群落的影响至关重要[26]。研究表明土壤pH和电导率是表征盐碱土盐碱化程度的重要参数，其数值与土壤盐分含量具有显著的相关性[27]。不同盐分含量对土壤微生物群落结构具有重要影响，随着土壤盐分增加土壤结构会变得更加紧密，影响土壤的透气性和排水性等，进而影响土壤微生物的生存[17]。关于土壤盐度对AOB的影响，目前的研究结果因环境因子不同而不一致。研究表明AOB群落多样性会随着土壤盐度的增加而降低[28]。也有研究表明在一定的盐度范围内，随着土壤盐度增加AOB群落的多样性是上升的，过高的盐度才会使多样性降低[29]。此外，有研究表明Nitrosomonas是盐碱环境中AOB的主要类群[30]，这与本研究结果一致。本研究中，高盐土壤中AOB群落的优势类群为Nitrosomonas，同时高盐土壤中AOB群落的多样性指数也高于低盐土壤。虽然高盐会在一定程度上抑制AOB群落的生长，但盐度对AOB群落结构具有选择性作用，抑制少数相对丰度高的AOB，富集多数相对丰度低的AOB，从而提高了AOB群落的丰富度和多样性[31]。
4 结论
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添加DMPP后土壤AOB多样性、群落组成和群落组装过程发生显著变化。DMPP通过抑制AOB群落中少数高丰度优势类群的生长，降低其相对丰度，进而缓解资源竞争压力并释放生态位空间，这为低丰度AOB类群的增殖创造了条件，而低丰度类群的丰度增加最终提高了AOB群落的多样性。高相对丰度类群AOB相对丰度降低是DMPP抑制硝化作用过程的主要原因。DMPP处理使AOB群落组装过程中随机性过程增加，确定性过程降低，随机性过程能够使微生物群落产生更多样化的生态功能，提供更高的微生物多样性，可部分缓解DMPP对AOB群落造成的干扰，有利于维持土壤生态系统的多功能性。DMPP在高盐土壤中抑制效率更高，可考虑在保证有效硝化抑制的前提下适当降低DMPP的推荐施用量，低盐土壤则可能需要施用相对高盐土壤稍高的DMPP剂量。
作者贡献声明
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周家庆：论文构思、数据分析、撰写文章、修订审阅；徐敏：数据分析、撰写文章；汪峰：获取基金、提供资源；刘浩：软件程序分析、数据分析；朱诗君：提供资源；曹慧：获取基金、项目管理、论文审阅。
作者利益冲突公开声明
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作者声明不存在任何可能会影响本文所报告工作的已知经济利益或个人关系。
基金
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  • 国家自然科学基金(32471725)
  • 宁波市“科技创新2025”重大专项(2022Z169)
  • 宁波市“科技创新2025”重大专项(2023Z114)
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2025年第65卷第12期
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文章信息
doi: 10.13343/j.cnki.wsxb.20250449
  • 接收时间：2025-06-09
  • 首发时间：2025-12-08
  • 出版时间：2025-12-04
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  • 收稿日期：2025-06-09
  • 录用日期：2025-08-25
基金
National Natural Science Foundation of China(32471725)
国家自然科学基金(32471725)
Major Special Projects of “Science and Technology Innovation 2025” in Ningbo(2022Z169)
宁波市“科技创新2025”重大专项(2022Z169)
Major Special Projects of “Science and Technology Innovation 2025” in Ningbo(2023Z114)
宁波市“科技创新2025”重大专项(2023Z114)
作者信息
    1.南京农业大学 生命科学学院，农业农村部农业环境微生物重点实验室，江苏 南京
    2.宁波市农业科学研究院，宁波市特色农产品质量安全检测与控制重点实验室，浙江 宁波

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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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