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施肥模式对青稞田土壤潜在固氮速率和自生固氮微生物群落结构的影响

马瑞萍 戴相林 刘国一 谢永春 高小丽 高雪

马瑞萍, 戴相林, 刘国一, 谢永春, 高小丽, 高雪. 施肥模式对青稞田土壤潜在固氮速率和自生固氮微生物群落结构的影响[J]. 中国生态农业学报(中英文), 2021, 29(10): 1692−1703 doi: 10.13930/j.cnki.cjea.210148
引用本文: 马瑞萍, 戴相林, 刘国一, 谢永春, 高小丽, 高雪. 施肥模式对青稞田土壤潜在固氮速率和自生固氮微生物群落结构的影响[J]. 中国生态农业学报(中英文), 2021, 29(10): 1692−1703 doi: 10.13930/j.cnki.cjea.210148
MA R P, DAI X L, LIU G Y, XIE Y C, GAO X L, GAO X. Effects of fertilizer patterns on the potential nitrogen fixation rate and community structure of asymbiotic diazotroph in highland barley fields on the Tibetan Plateau[J]. Chinese Journal of Eco-Agriculture, 2021, 29(10): 1692−1703 doi: 10.13930/j.cnki.cjea.210148
Citation: MA R P, DAI X L, LIU G Y, XIE Y C, GAO X L, GAO X. Effects of fertilizer patterns on the potential nitrogen fixation rate and community structure of asymbiotic diazotroph in highland barley fields on the Tibetan Plateau[J]. Chinese Journal of Eco-Agriculture, 2021, 29(10): 1692−1703 doi: 10.13930/j.cnki.cjea.210148

施肥模式对青稞田土壤潜在固氮速率和自生固氮微生物群落结构的影响

doi: 10.13930/j.cnki.cjea.210148
基金项目: 省部共建青稞和牦牛种质资源与遗传改良国家重点实验室自主课题(XZNKY-2020-C-007Z09)和西藏自治区自然科学基金项目(XZ2019ZRG-98)资助
详细信息
    作者简介:

    马瑞萍, 研究方向为养分资源高效利用。E-mail: marp0825@126.com

    通讯作者:

    戴相林, 主要研究方向为养分资源高效利用。E-mail: dxlok911@163.com

  • 中图分类号: S154.3

Effects of fertilizer patterns on the potential nitrogen fixation rate and community structure of asymbiotic diazotroph in highland barley fields on the Tibetan Plateau

Funds: This study was supported by the State Key Laboratory of Hulless Barley and Yak Germplasm Resources and Genetic Improvement (XZNKY-2020-C-007Z09), and the Natural Science Foundation of Tibet Autonomous Region (XZ2019ZRG-98)
More Information
  • 摘要: 自生固氮是陆地生态系统氮素的重要来源。本研究以位于西藏拉萨的肥料长期定位试验站为平台, 选取不施肥(CK)、单施化肥(F)、单施羊粪(M)、化肥配施羊粪(FM)和化肥配施秸秆(FS)5个处理, 于青稞收获期采集土样, 采用15N同位素标记、定量PCR和高通量测序技术, 分析不同施肥模式下土壤理化性质、固氮微生物丰度、群落结构和潜在固氮速率的变化规律, 以期为西藏高原青稞田制定科学合理的施肥策略, 保障西藏高原农业绿色发展提供科学依据。结果表明: 1)相比于CK和F处理, M、FM和FS处理可显著(P<0.05)提高土壤有机碳和全氮含量。FM处理下土壤铵态氮含量最高, 且显著高于其他施肥处理(P<0.05), 而M处理下土壤有机碳、全氮、硝态氮、有效磷和速效钾含量最高, 且显著高于其他施肥处理(P<0.05)。2)不同施肥处理下青稞田土壤潜在固氮速率为2.63~4.07 μg∙kg−1∙d−1。施肥会降低土壤潜在固氮速率, 增施有机肥(羊粪或秸秆)则加剧了抑制效应, 但羊粪的抑制效应小于秸秆, 土壤铵态氮含量是影响土壤潜在固氮速率的主要因子。3) M或FM处理可显著(P<0.05)提高nifH基因丰度, 而F或FS处理则相反, 土壤全氮含量是影响nifH基因丰度的关键因子。4)不同施肥模式显著改变了固氮微生物群落结构, 其群落结构相似性大致可以分为3类, 分别为CK、M以及施用化肥处理(F、FM和FS), 有效磷、pH和C/N是调控固氮微生物群落结构的关键因子。综上所述, 单施羊粪(M)处理是提高青稞田土壤肥力、增加固氮微生物丰度, 减少固氮速率下降幅度的最佳施肥模式。
  • 图  1  不同施肥模式下固氮微生物丰度和土壤潜在固氮速率的变化

    CK: 不施肥; F: 施氮磷钾化肥; M: 单施羊粪; FM: 氮磷钾化肥+羊粪; FS: 氮磷钾化肥+秸秆。不同字母表示同一指标处理之间存在显著性差异(P<0.05)。CK: without fertilizer; F: only mineral NPK fertilizers; M: only sheep manure; FM: chemical NPK fertilizers plus sheep manure; FS: chemical NPK fertilizers plus barley straw. Different letters for the indicator indicate significant differences among fertilizer treatments at P<0.05 level.

    Figure  1.  Changes of diazotrophic abundance and potential N fixation rate under different fertilizer regimes of hulless barley field

    图  2  土壤理化性质与固氮微生物丰度和固氮潜势之间的关系

    图A表示土壤理化性质与固氮微生物丰度和固氮潜势之间的Spearman相关性。其中蓝色和红色圆圈分别表示指标间的正相关和负相关, 圆圈大小表示指标间相关性的高低, “×”表示指标间相关性不显著。图B和图C分别为基于逐步回归分析揭示影响固氮微生物丰度和潜在固氮速率的关键土壤因子。其中, 蓝色线表示回归线, 灰色背景表示95%的置信区间。CK: 不施肥; F: 施氮磷钾化肥; M: 单施羊粪; FM: 氮磷钾化肥+羊粪; FS: 氮磷钾化肥+秸秆。Figure A shows the relationships among soil properties, diazotrophic abundance and potential N fixation rate (A) according to Spearman’s correlation coefficient, in which, blue and red circles denote positive and negative correlations, respectively, large to small circles indicate high to low correlations. “×” in circles means no significant correlaiton between indexes. SOC: soil organic carbon; TN: total nitrogen; SWC: soil water content; AP: available phosphorus; AK: available potassium; PNFR: potential N fixation rate. Figure B and C show the main predictors responsible for the changes in the diazotrophic abundance and PNFR based on stepwise linear regression analysis, respectively. The grey background indicates a 95% confidence interval. CK: without fertilizer; F: only mineral NPK fertilizers; M: only sheep manure; FM: chemical NPK fertilizers plus sheep manure; FS: chemical NPK fertilizers plus barley straw.

    Figure  2.  Relationships among soil properties, diazotrophic abundance and potential N fixation rate

    图  3  不同施肥模式下固氮微生物纲水平(A)和属水平(B)(相对丰度>1%)组成变化

    CK: 不施肥; F: 施氮磷钾化肥; M: 单施羊粪; FM: 氮磷钾化肥+羊粪; FS: 氮磷钾化肥+秸秆。不同小写字母表示不同处理间差异显著(P<0.05)。柱体上无小写字母表示各处理间无显著差异。CK: without fertilizer; F: only mineral NPK fertilizers; M: only sheep manure; FM: chemical NPK fertilizers plus sheep manure; FS: chemical NPK fertilizers plus barley straw. Different lowercase letters indicate significant differences among treatments at P<0.05. No lowercase letters indicate significant differences among treatments at P>0.05.

    Figure  3.  Changes of composition of diazotroph at the phylum (A) and genus (B) levels (relative abundance > 1%) under different fertilizer regimes

    图  4  主坐标分析不同施肥模式对固氮微生物群落结构的影响

    CK: 不施肥; F: 施氮磷钾化肥; M: 单施羊粪; FM: 氮磷钾化肥+羊粪; FS: 氮磷钾化肥+秸秆。CK: without fertilizer; F: only mineral NPK fertilizer; M: only sheep manure; FM: chemical NPK fertilizers plus sheep manure; FS: chemical NPK fertilizers plus barley straw.

    Figure  4.  Principal coordinate analysis (PCoA) based on Bray-Curtis dissimilarity of diazotrophic communities under different fertilizer regimes

    图  5  典范对应分析土壤理化因子(膨胀因子<10)与固氮微生物群落结构的关系

    AP代表有效磷。CK: 不施肥; F: 施氮磷钾化肥; M: 单施羊粪; FM: 氮磷钾化肥+羊粪; FS: 氮磷钾化肥+秸秆。AP represents available phosphorus. CK: without fertilizer; F: only mineral NPK fertilizer; M: only sheep manure; FM: chemical NPK fertilizers plus sheep manure; FS: chemical NPK fertilizers plus barley straw.

    Figure  5.  Relationship between soil properties (VIF<10) and diazotrophic communities based on canonical correlation analysis (CCA)

    表  1  不同施肥模式下青稞田土壤理化性质的变化

    Table  1.   Changes of soil physicochemical properties under different fertilizer regimes of hulless barley field

    理化性质 Physicochemical propertyCKFMFMFS
    有机碳 Organic carbon (g∙kg−1) 11.7±0.04d 12.2±0.02d 18.6±0.02a 15.1±0.03b 12.8±0.03c
    全氮 Total nitrogen (g∙kg−1) 1.24±0.00d 1.21±0.00d 1.82±0.00a 1.54±0.00b 1.32±0.00c
    碳氮比 C/N 9.50±0.33c 10.08±0.09ab 10.22±0.15a 9.86±0.26abc 9.71±0.29bc
    土壤含水量 Water content (%) 12.17±0.06e 13.62±0.16b 14.30±0.07a 13.24±0.03c 12.86±0.09d
    pH 8.44±0.02a 8.25±0.01b 8.25±0.01b 8.10±0.02d 8.20±0.02c
    铵态氮 NH4+-N (mg∙kg−1) 11.82±1.21cd 11.05±1.30d 13.22±0.64bc 16.16±0.02a 13.67±0.83b
    硝态氮 NO3-N (mg∙kg−1) 1.88±0.15c 3.30±0.16b 5.00±0.50a 4.97±0.17a 3.28±0.08b
    有效磷 Available phosphorus (mg∙kg−1) 23.93±1.28d 45.18±1.89bc 49.65±2.34a 48.02±1.24ab 44.10±0.92c
    速效钾 Available potassium (mg∙kg−1) 65.17±1.75d 74.35±2.67c 120.71±5.87a 92.33±4.75b 72.84±1.31c
      CK: 不施肥; F: 施氮磷钾化肥; M: 单施羊粪; FM: 氮磷钾化肥+羊粪; FS: 氮磷钾化肥+秸秆。同行不同字母表示处理之间存在显著性差异(P<0.05)。CK: without fertilizer; F: only mineral NPK fertilizers; M: only sheep manure; FM: chemical NPK fertilizers plus sheep manure; FS: chemical NPK fertilizers plus barley straw. Different letters in the same row indicate significant differences among fertilizer treatments at P<0.05 level.
    下载: 导出CSV

    表  2  土壤理化性质与固氮菌优势纲相对丰度之间的相关性分析

    Table  2.   Spearman correlation between relative abundance of dominated class of diazotroph and soil physicochemical properties

    纲 Class有机碳
    Organic
    carbon
    全氮
    Total
    nitrogen
    碳氮比
    C/N
    pH铵态氮
    NH4+-N
    硝态氮
    NO3-N
    有效磷
    Available
    phosphorus
    速效钾
    Available
    potassium
    含水量
    Water
    content
    β-变形菌纲
    Betaproteobacteria
    −0.483 −0.414 −0.811** −0.196 0.111 −0.484 −0.439 −0.554* −0.657**
    α-变形菌纲
    Alphaproteobacteria
    0.470 0.448 0.496 −0.174 0.121 0.559* 0.371 0.568* 0.607*
    γ-变形菌纲
    Gammaproteobacteria
    0.841** 0.800** 0.661** −0.326 0.429 0.751** 0.789** 0.882** 0.671**
    δ-变形菌纲
    Deltaproteobacteria
    −0.329 −0.448 −0.193 0.149 −0.154 −0.504 −0.493 −0.525* −0.679**
    丰佑菌纲 Opitutae −0.459 −0.403 −0.559* 0.148 −0.236 −0.583* −0.608* −0.652** −0.685**
      *和**分别表示P<0.05和P<0.01水平显著相关。* and ** mean significant correlation at P<0.05 and P<0.01 levels, respectively.
    下载: 导出CSV

    表  3  土壤理化性质与固氮菌优势属相对丰度之间的相关性分析

    Table  3.   Spearman correlation between relative abundance of dominated genera of diazotroph and soil physicochemical properties

    属 Genus有机碳
    Organic
    carbon
    全氮
    Total
    nitrogen
    碳氮比
    C/N
    pH铵态氮
    NH4+-N
    硝态氮
    NO3-N
    有效磷
    Available
    phosphorus
    速效钾
    Available
    potassium
    含水量
    Water
    conten
    地杆菌属 Geobacter −0.360 −0.420 −0.186 0.106 −0.132 −0.468 −0.454 −0.493 −0.654**
    固氮弧菌属 Azoarcus −0.624* −0.577* −0.671** 0.289 −0.189 −0.718** −0.707** −0.782** −0.896**
    假食酸菌属 Pseudacidovorax −0.528* −0.459 −0.782** −0.203 0.061 −0.500 −0.371 −0.504 −0.568*
    红长命菌属 Rubrivivax −0.308 −0.358 −0.250 −0.335 −0.107 −0.375 −0.246 −0.279 −0.357
    脱氮单孢菌属 Dechloromonas −0.678** −0.711** −0.471 0.303 −0.432 −0.708** −0.454 −0.614 −0.582*
    动胶菌属 Zoogloea −0.335 −0.358 −0.564* −0.113 0.021 −0.340 −0.361 −0.464 −0.489
    念珠藻属 Nostoc 0.303 0.266 0.626* 0.167 −0.097 0.418 0.429 0.500 0.695**
    慢生根瘤菌属 Bradyrhizobium 0.556* 0.560* 0.511 0.018 0.154 0.624* 0.411 0.596* 0.554*
    Azohydromonas 0.156 0.078 0.375 −0.413 0.071 0.239 0.254 0.329 0.364
    鱼腥藻属 Anabaena 0.458 0.420 0.650** −0.011 −0.004 0.483 0.600* 0.698** 0.704**
    贪噬菌属 Variovorax −0.413 −0.325 −0.182 0.317 −0.157 −0.263 −0.250 −0.186 −0.254
    固氮螺菌属 Azospirillum 0.215 0.154 0.525* −0.031 −0.195 0.324 0.404 0.463 0.656**
    甲基细菌属 Methylobacter 0.891** 0.865** 0.600** −0.425 0.500 0.842** 0.879** 0.918** 0.768**
    Skermanella 0.345 0.319 0.407 −0.366 0.264 0.456 0.411 0.461 0.550*
      *和**分别表示P<0.05和P<0.01水平显著相关。* and ** mean significant correlation at P<0.05 and P<0.01 levels, respectively.
    下载: 导出CSV
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  • 收稿日期:  2021-03-12
  • 录用日期:  2021-04-28
  • 网络出版日期:  2021-08-13
  • 刊出日期:  2021-10-01

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