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

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

左明雪^{1, 2,},
孙杰²,
徐如玉^{1, 2},
袁银龙^{1, 2},
顾文杰²,
卢钰升²,
徐培智^{1, 2, ,},
解开治^{1, 2, ,}

1.
甘肃农业大学资源与环境学院，甘肃　兰州　730070
2.
广东省农业科学院农业资源与环境研究所，广东　广州　510640

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
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- 被引次数: 0
出版历程
- 收稿日期: 2020-05-06
- 修回日期: 2020-07-07
- 刊出日期: 2020-09-28

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

ZUO Mingxue^{1, 2
,},
SUN Jie²,
XU Ruyu^{1, 2},
YUAN Yinlong^{1, 2},
GU Wenjie²,
LU Yusheng²,
XU Peizhi^{1, 2
, ,},
XIE Kaizhi^{1, 2
, ,}

1.
College of Resources and Environmental Science, Gansu Agricultural University, Lanzhou, Gansu　730070, China
2.
Institute of Agricultural Resources and Environment, Guangdong Academy of Agricultural Sciences, Guangzhou, Guangdong　510640, China

摘要

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）NiR、narB、nasA、nirA、nirB、napA、nrfA、nifH、ureC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（NiR、narB、nasA、nirA、nirB）、异化氮还原基因（napA、nrfA）、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。
- 土壤氮循环 /
- 氮素转化 /
- 变形球囊霉 /
- 微生物群落多样性 /
- 功能基因芯片
Abstract: Objective Effects of AMF and organic fertilizer on nitrogen (N) transformation and microbial N-cycling gene in rhizosphere soil at sweet corn field were studied to improve the fertilization practice. Method In a field experiment, sweet corn was planted under various fertilization treatments, and the N-transformation between the plants and the soil monitored. The high-throughput sequencing platform, GeoChip 5.0, was used to determine the microbial community structure and N-cycling genes. Upon the base fertilization of P₂O₅150 kg·hm^-2 and K₂O 225 kg·hm^-2, 7 modifications were applied with 3 replicates for the soil treatments: (1) no N addition (CK), (2) optimized fertilization (OF), (3) organic N replacing 10% chemical N (ORF10), (4) organic N replacing 20% chemical N (ORF20), (5) ORF10 inoculated with Glomus versiforme (ORF10+AMF), (6) ORF20 inoculated with G. versiforme (ORF20+AMF), and, (7) CK inoculated with G. versiforme (CK+AMF). The physical and chemical analyses were performed on the plant and soil samples, and GeoChip 5.0 analyzed the community structure and N-cycling genes of the microbes in the rhizosphere soil under different treatments. Result The inoculation of G. versiforme in soil significantly increased the N utilization efficiency by the sweet corn plants as well as the activity of N metabolizing enzymes in the rhizosphere microorganisms. When AMF inoculation combined with organic N fertilization, significant effects were observed on increases of the efficiency of N fertilizer (NAE), partial productivity of N fertilizer (PFP), and absorption and utilization efficiency of N fertilizer (NRE) between the plants and the soil, as well as the activities of nitrate reductase (NR), glutamic acid synthetase (GOGAT), and glutamine synthetase (GS) in the microbes. Among all treatments, ORF20+AMF improved the N utilization by the plants the greatest—NAE, PFP, NRE, and NPE rose by 31.15%, 28.08%, 6.95%, and 10.41%, respectively, over those under OF. The relative signal intensities of NiR, narB, nasA, nirA, nirB, napA, nrfA, nifH, and ureC in ORF10+AMF or ORF20+AMF were significantly higher than those in ORF10 or ORF20. That of hzo in ORF 20+AMF, in contrast, decreased by 20% compared with ORF20, which inviably caused a decreased N release by ways of N₂. Conclusion The presence of AMF in soil enhanced the relative signal intensities of assimilating nitrogen reducing gene NiR, narB, nasA, nirA, and nirB, the N-reducing napA and nrfA , N-fixing nifH , ammonifying ureC, improved the NAE, PFP, and NRE, and increased the microbial N metabolizing enzyme activities. Meanwhile, the declined signal intensities on nitrifying hao, and ammoxidating hzo augmented the N-transformation from soil to plant. The application of AMF and organic fertilizer could, hence, be a promising approach to mitigate the dependency on chemical N fertilizer while promoting the crop yield of sweet corn in the field.
- nitrogen cycle /
- nitrogen transformation /
- Glomus versiforme /
- rhizosphere microbial community /
- GeoChip

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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

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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

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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

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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.

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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

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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）
处理 Treatment	N	P₂O₅	K₂O	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

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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/%
CK	7.13±0.32 c	58.00±6.00 c	22.50±0.64 e
OF	12.98±0.39 c	64.67±4.73 bc	26.92±0.37 d
ORF10	11.58±0.24 c	65.33±4.16 bc	28.87±0.76 d
ORF20	7.63±1.29 c	66.00±2.65 bc	28.65±2.65 d
ORF10+AMF	19.64±3.99 b	74.67±9.29 ab	41.56±2.02 b
ORF20+AMF	29.25±8.17 a	68.67±1.15 b	38.13±3.18 c
CK+AMF	24.53±0.45 ab	81.00±5.00 a	45.34±0.56 a
显著性 Significance	P值 P Value
AMF	＜0.001	＜0.001	＜0.001
有机无机配施（ORF）	0.378	0.668	0.282
AMF×ORF	0.025	0.019	＜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.

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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	-	-	-	-
OF	27.31±2.36 b	49.39±1.79 b	37.80±2.23 b	72.39±7.17 ab
ORF10	19.74±2.89 c	44.28±2.17 c	31.23±1.92 c	63.00±5.22 b
ORF20	23.98±1.05 bc	51.58±0.20 b	33.64±1.92 bc	71.53±7.04 ab
CK+AMF	-	-	-	69.46±12.12 a
ORF10+AMF	26.67±3.32 b	51.21±2.89 b	36.39±1.58 bc	73.16±6.90 ab
ORF20+AMF	35.53±2.75 a	63.13±3.13 a	44.75±5.59 a	79.82±6.32 a
显著性 Significance	P值 P Value
AMF	＜0.001	＜0.001	0.002	0.037
有机无机配施（ORF）	0.003	＜0.001	0.020	0.074
AMF×ORF	0.170	0.133	0.147	0.807

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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）
CK	11.04±0.74 c	7.13±0.40 b	5.30±0.12 b	82.05±0.62 b
OF	11.30±1.17 c	9.19±0.66 a	6.58±0.43 a	82.63±8.54 b
ORF10	18.04±1.40 a	8.30±1.11 ab	5.27±0.46 bc	91.55±8.04 b
ORF20	15.49±1.26 b	8.37±1.52 ab	4.66±0.53 bc	102.29±3.93 a
ORF10+AMF	10.64±1.26 c	7.71±0.59 ab	4.62±0.33 c	92.20±4.21 b
ORF20+AMF	5.55±0.64 d	8.02±0.55 ab	6.06±0.25 a	82.24±5.64 b
CK+AMF	10.07±1.13 c	8.56±0.68 ab	3.79±0.19 c	41.20±1.06 c
显著性 Significance	P 值P Value
AMF	＜0.001	0.704	0.149	＜0.001
有机无机配施（ORF）	＜0.001	0.793	0.006	＜0.001
AMF×ORF	＜0.001	0.145	＜0.001	＜0.001

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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R	P值 P value
MRPP	δ=0.169	＜0.001
Anosim	R=0.979	＜0.001
Adonis	R=0.859	＜0.001

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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

doi: 10.19303/j.issn.1008-0384.2020.09.013

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）

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

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留言板

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

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）

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

计量

出版历程

目录

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）