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丛枝菌根真菌与有机肥配施对甜玉米根际土壤氮素转化及氮循环微生物功能基因的影响

左明雪 孙杰 徐如玉 袁银龙 顾文杰 卢钰升 徐培智 解开治

左明雪,孙杰,徐如玉,等. 丛枝菌根真菌与有机肥配施对甜玉米根际土壤氮素转化及氮循环微生物功能基因的影响 [J]. 福建农业学报,2020,35(9):1012−1025 doi: 10.19303/j.issn.1008-0384.2020.09.013
引用本文: 左明雪,孙杰,徐如玉,等. 丛枝菌根真菌与有机肥配施对甜玉米根际土壤氮素转化及氮循环微生物功能基因的影响 [J]. 福建农业学报,2020,35(9):1012−1025 doi: 10.19303/j.issn.1008-0384.2020.09.013
ZUO M X, SUN J, XU R Y, et al. Effects of AMF and Organic Fertilizer on N-transformation and Microbial N-cycling Genes in Rhizosphere Soil of Sweet Corn Field [J]. Fujian Journal of Agricultural Sciences,2020,35(9):1012−1025 doi: 10.19303/j.issn.1008-0384.2020.09.013
Citation: ZUO M X, SUN J, XU R Y, et al. Effects of AMF and Organic Fertilizer on N-transformation and Microbial N-cycling Genes in Rhizosphere Soil of Sweet Corn Field [J]. Fujian Journal of Agricultural Sciences,2020,35(9):1012−1025 doi: 10.19303/j.issn.1008-0384.2020.09.013

丛枝菌根真菌与有机肥配施对甜玉米根际土壤氮素转化及氮循环微生物功能基因的影响

doi: 10.19303/j.issn.1008-0384.2020.09.013
基金项目: 国家重点研发计划项目(2018YFD0200706);广东省科技计划项目(2016A030313776、2017B020233002、2017B020203002);广州市珠江科技新星专项(201710010182);广东省农业科学院十三五学科团队建设项目(201801XX)
详细信息
    作者简介:

    左明雪(1995−),女,硕士,主要从事土壤氮循环方面研究(E-mail:17808322297@163.com

    通讯作者:

    徐培智(1963-),男,研究员,主要从事植物营养与高效施肥研究(E-mail:pzxu007@163.com

    解开治(1977−),男,博士,研究员,主要从事植物营养研究(E-mail:xiekzgsau@163.com

  • 中图分类号: S 436.681;S 476

Effects of AMF and Organic Fertilizer on N-transformation and Microbial N-cycling Genes in Rhizosphere Soil of Sweet Corn Field

  • 摘要:   目的  研究丛枝菌根真菌(Arbuscular mycorrhizal fungi, AMF)配施有机肥对甜玉米根际土壤氮素转化及氮循环微生物功能基因的影响,明晰AMF配施有机肥对甜玉米-土壤氮循环的微生物学过程,为氮肥利用率提高、化学氮肥减施增效提供技术支撑。  方法  采用大田区组试验,在磷(P2O5 150 kg·hm-2)、钾(K2O 225 kg·hm-2)施肥水平一致的基础上设计7个施肥处理:(1)不施氮肥(CK);(2)优化施肥(OF);(3)有机氮肥替代10%化学氮肥(ORF10);(4)有机氮肥替代20%化学氮肥(ORF20);(5)有机氮肥替代10%化学氮肥+增施变形球囊霉(Glomus versiforme)(ORF10+AMF);(6)有机氮肥替代20%化学氮肥+增施变形球囊霉(ORF20+AMF);(7)不施氮肥+增施变形球囊霉(CK+AMF);每个处理3次重复。应用土壤常规理化指标分析方法和微生物功能基因芯片(GeoChip 5.0)技术,对不同施肥处理的甜玉米氮素利用率、氮代谢相关酶活性及氮循环功能基因进行分析。  结果  增施变形球囊霉能显著提高氮利用率。变形球囊霉配施有机肥对氮肥农学效率(NAE)、氮肥偏生产力(PFP)、氮肥吸收利用率(NRE)、硝酸还原酶(NR)、谷氨酸合酶(GOGAT)和谷氨酰胺合成酶(GS)活性具有极显著的正交互效应(P<0.001)。在所有处理中ORF20+AMF处理土壤氮素利用效率最高,农学效率(NAE)、氮肥偏生产力(PFP)、氮肥吸收利用率(NRE)、氮素生理利用率(NPE)较OF处理分别提高31.15%、28.08%、6.95%、10.41%。在氮循环微生物功能基因中,增施变形球囊霉处理组(ORF10+AMF、ORF20+AMF)NiRnarBnasAnirAnirBnapAnrfAnifHureC基因相对强度显著高于对应未施菌处理组(ORF10、ORF20),ORF20+AMF处理的氨氧化基因hzo相对丰度比ORF20处理降低了20%,减少了氮素由N2释放途径损失的可能。  结论  增施变形球囊霉配施有机肥可显著调增同化氮还原基因(NiRnarBnasAnirAnirB)、异化氮还原基因(napAnrfA)、氮固定基因(nifH)、氨化作用基因(ureC)相对强度,降低硝化基因(hao)和氨氧化基因(hzo)相对丰度,驱动土壤氮素循环向植物氮高效利用的途径转化,提高氮肥农学效率(NAE)、氮肥偏生产力(PFP)、氮肥吸收利用率(NRE)和根系氮代谢酶活性,实现甜玉米化学氮肥减施增效的目标。
  • 图  1  不同处理亚类氮循环功能基因标准化相对信号强度

    Figure  1.  Relative signal intensity of normalized microbial N-cycling genes under treatments

    图  2  各处理土壤微生物氮代谢功能基因结构DCA排序图

    Figure  2.  DCA ordination diagram on structure of N-metabolizing genes in soil microorganisms

    图  3  不同施肥处理氮循环关键功能基因标准化相对信号强度

    Figure  3.  Normalized relative signal intensity of microbial N-cycling genes under treatments

    图  4  不同施肥处理间氮代谢功能基因丰度的变化

    注:括号中指示了根际土N基因丰度的百分比变化。以增施变形球囊霉处理为准,基因丰度增加用红色标记;降低用绿色标记;其中*、**、***分别表示在0.05、0.01、0.001水平存在显著差异。此处使用的GeoChip版本没有将灰色基因作为目标,也没有发现或未发现未注释。

    Figure  4.  Changes in abundance of N-metabolizing genes in soil microbes under treatments

    Note: The percentage change of N gene abundance in rhizosphere soil is indicated in parentheses. Subject to the treatment of increased application of cystic mildew, the gene abundance is increased and marked with red; the decrease is marked with green; the *, **, *** indicate significant difference at 0.05, 0.01, 0.001 level, respectively. The version of GeoChip used here does not target the gray gene, nor has it been found or found. Found uncommented.

    图  5  氮循环功能基因和理化性质的RDA分析

    Figure  5.  RDA analysis on microbial N-cycling genes and physicochemical properties of soil under treatments

    表  1  不同处理间施肥方案

    Table  1.   Fertilization treatments

    处理
    Treatment
    施肥用量 Fertilization schemes/(kg·hm−2
    NP2O5K2O有机肥 Organic fertilizer
    CK,不施氮肥 0 150 225 0
    OF,优化施肥 330 150 225 0
    ORF10,有机氮肥替代10%化学氮肥 297 150 225 1650
    ORF20,有机氮肥替代20%化学氮肥 264 150 225 3300
    ORF10+AMF,有机氮肥替代10%化学氮肥+增施变形球囊霉(G. versiforme 297 150 225 1650
    ORF20+AMF,有机氮肥替代20%化学氮肥+增施变形球囊霉(G. versiforme 264 150 225 3300
    CK+AMF,不施氮肥+增施变形球囊霉(G. versiforme 0 150 225 0
    下载: 导出CSV

    表  2  不同施肥处理对甜玉米孢子量、侵染率、侵染密度的影响

    Table  2.   Spore quantity, infection rate, and infection density on sweet corn under treatments

    处理 Treatment孢子密度 Spore density/(个·g−1侵染率 Colonization/%侵染强度 Colonization intensity/%
    CK7.13±0.32 c58.00±6.00 c22.50±0.64 e
    OF12.98±0.39 c64.67±4.73 bc26.92±0.37 d
    ORF1011.58±0.24 c65.33±4.16 bc28.87±0.76 d
    ORF207.63±1.29 c66.00±2.65 bc28.65±2.65 d
    ORF10+AMF19.64±3.99 b74.67±9.29 ab41.56±2.02 b
    ORF20+AMF29.25±8.17 a68.67±1.15 b38.13±3.18 c
    CK+AMF24.53±0.45 ab81.00±5.00 a45.34±0.56 a
    显著性 SignificancePP Value
    AMF<0.001<0.001<0.001
    有机无机配施(ORF)0.3780.6680.282
    AMF×ORF0.0250.019<0.001
    注:同列数据后不同小写字母表示处理间差异显著性(P<0.05)。数据为均值±标准误。
    Note: Different lowercase in each column indicates significant difference among different treatments(P<0.05). Values represent the mean±standard error.
    下载: 导出CSV

    表  3  不同施肥处理甜玉米氮利用率

    Table  3.   Nutilization efficiency of sweet corn under treatments

    处理 Treatment氮肥农学效率 NAE/(kg·kg−1氮肥偏生产力 PFP/(kg·kg−1氮肥吸收利用率 NRE/%氮素生理利用率 NPE/(kg·kg−1
    CK----
    OF27.31±2.36 b49.39±1.79 b37.80±2.23 b72.39±7.17 ab
    ORF1019.74±2.89 c44.28±2.17 c31.23±1.92 c63.00±5.22 b
    ORF2023.98±1.05 bc51.58±0.20 b33.64±1.92 bc71.53±7.04 ab
    CK+AMF---69.46±12.12 a
    ORF10+AMF26.67±3.32 b51.21±2.89 b36.39±1.58 bc73.16±6.90 ab
    ORF20+AMF35.53±2.75 a63.13±3.13 a44.75±5.59 a79.82±6.32 a
    显著性 SignificancePP Value
    AMF<0.001<0.0010.0020.037
    有机无机配施(ORF)0.003<0.0010.0200.074
    AMF×ORF0.1700.1330.1470.807
    下载: 导出CSV

    表  4  不同施肥处理甜玉米根内系氮代谢相关的酶活性

    Table  4.   Activity of microbial N metabolizing enzymes in sweet corn under treatments

    处理
    Treatment
    硝酸还原酶
    NR/(U·g−1
    亚硝酸还原酶
    NiR/(μmol·h−1·g−1
    谷氨酰胺合成酶
    GS/(U·g−1
    谷氨酸合酶
    GOGAT/(nmol·min−1·g−1
    CK11.04±0.74 c7.13±0.40 b5.30±0.12 b82.05±0.62 b
    OF11.30±1.17 c9.19±0.66 a6.58±0.43 a82.63±8.54 b
    ORF1018.04±1.40 a8.30±1.11 ab5.27±0.46 bc91.55±8.04 b
    ORF2015.49±1.26 b8.37±1.52 ab4.66±0.53 bc102.29±3.93 a
    ORF10+AMF10.64±1.26 c7.71±0.59 ab4.62±0.33 c92.20±4.21 b
    ORF20+AMF5.55±0.64 d8.02±0.55 ab6.06±0.25 a82.24±5.64 b
    CK+AMF10.07±1.13 c8.56±0.68 ab3.79±0.19 c41.20±1.06 c
    显著性 SignificancePP Value
    AMF<0.0010.7040.149<0.001
    有机无机配施(ORF)<0.0010.7930.006<0.001
    AMF×ORF<0.0010.145<0.001<0.001
    下载: 导出CSV

    表  5  不同施肥处理间微生物氮循环功能基因不相似性检验

    Table  5.   Dissimilarity test on microbial N-cycling genes under treatments

    不相似性检验
    Dissimilarity test
    δ或RP
    P value
    MRPP δ=0.169 <0.001
    Anosim R=0.979 <0.001
    Adonis R=0.859 <0.001
    下载: 导出CSV
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  • 收稿日期:  2020-05-06
  • 修回日期:  2020-07-07
  • 刊出日期:  2020-09-28

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