秸秆还田下土壤碳氮分布与固氮和反硝化细菌种群的相互影响

doi:10.3969/j.issn.1004-1524.20240828

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (10): 2150-2164.DOI: 10.3969/j.issn.1004-1524.20240828

秸秆还田下土壤碳氮分布与固氮和反硝化细菌种群的相互影响

王云龙(), 贾生强, 崔玲宇, 吕豪豪, 沈阿林, 苏瑶()

浙江省农业科学院环境资源与土壤肥料研究所,浙江杭州 310021

收稿日期:2024-09-20 出版日期:2025-10-25 发布日期:2025-11-13
作者简介:王云龙(1979—),男,山东莱阳人,博士,副研究员,主要从事农业资源利用与产地环境治理研究。E-mail:wangyunlong@zju.edu.cn
通讯作者: *苏瑶,E-mail:stellasu@sina.com
基金资助:
国家小麦产业技术体系项目(CARS-3);桐庐县2024年受污染耕地安全利用关键技术研究与示范项目(SZQL-TL2024CS-002)

Interactions of soil carbon and nitrogen distribution with nitrogen fixing and denitrifying bacteria community under straw returning

WANG Yunlong(), JIA Shengqiang, CUI Lingyu, LYU Haohao, SHEN Alin, SU Yao()

Institute of Environment, Resource, Soil and Fertilizer, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2024-09-20 Online:2025-10-25 Published:2025-11-13

摘要/Abstract

摘要： 基于5年的定位试验,以秸秆不还田处理为对照(CK),研究秸秆还田后0~100 cm土层的有机碳和氮的组分分布,解析不同深度土层的生物固氮潜势和反硝化潜势、参与固氮和反硝化的主要微生物及其种群结构等的变化。结果表明,秸秆还田可较CK显著(p<0.05)增加0~60 cm土层的有机碳(OC)和颗粒有机碳(POC)含量,增幅分别为13.1%~243.8%、36.4%~143.8%,以及>20~60 cm土层的矿物结合态有机碳(MOC)含量,增幅为89.4%~272.9%,显著降低0~60 cm土层的溶解有机碳(DOC)含量,降幅在40.6%~75.9%。秸秆还田下,0~80 cm土层的总氮(TN)含量较CK显著增加14.3%~90.3%,但0~60 cm土层的碱解氮(AN)和硝态氮($\mathrm{NO}_{3}^{-}-\mathrm{N}$)含量显著下降16.9%~64.8%和12.9%~61.9%,0~40 cm土层的铵态氮($\mathrm{NH}_{4}^{+}-\mathrm{N}$)含量显著下降11.6%~24.8%,但>40~60 cm土层的$\mathrm{NH}_{4}^{+}-\mathrm{N}$含量显著增加至CK的1.4倍。与CK相比,秸秆还田下0~20 cm土层的生物固氮潜势显著增强25.2%,0~40 cm土层的反硝化潜势显著降低7.8%~82.0%。秸秆还田显著增加了土壤中nifH、nirS、nirK和nosZ基因的拷贝数,说明其促进了土壤中固氮细菌和反硝化细菌的生长。结构方程模型分析结果表明,秸秆还田后土壤DOC含量、土壤固氮细菌和反硝化细菌丰度的变化对土壤中$\mathrm{NO}_{3}^{-}-\mathrm{N}$和$\mathrm{NH}_{4}^{+}-\mathrm{N}$的含量与分布产生直接显著影响,其中,固氮细菌丰度的直接影响最大,对土壤$\mathrm{NO}_{3}^{-}-\mathrm{N}$和$\mathrm{NH}_{4}^{+}-\mathrm{N}$含量的影响效应分别为71.9%和-60.8%。

关键词: 秸秆还田, 土壤有机碳, 土壤矿质氮, 固氮细菌, 反硝化细菌

Abstract:

Based on a 5-year positioning test, the distribution of organic carbon and nitrogen components in soil from 0 to 100 cm depth after straw returning was studied, as well as the changes of soil biological nitrogen fixation and denitrification potential, abundance, diversity and community composition of main microorganisms involved in nitrogen fixation and denitrification, by taking the soil without straw returning as the control (CK). The results showed that straw returning significantly (p<0.05) increased soil organic carbon (OC) and particulate organic carbon (POC) content in 0-60 cm soil layer by 13.1%-243.8% and 36.4%-143.8%, respectively, and mineral-incorporated organic carbon (MOC) content in>20-60 cm soil layer by 89.4%-272.9% than those of CK, yet it significantly decreased soil dissolved organic carbon (DOC) content in 0-60 cm soil layer by 40.6%-75.9%. Under straw returning, the total nitrogen (TN) content in 0-80 cm soil layer was significantly increased by 14.3%-90.3% than that of CK, but the alkali-hydrolyzable nitrogen (AN) and nitrate nitrogen ($\mathrm{NO}_{3}^{-}-\mathrm{N}$) contents in 0-60 cm soil layer were decreased significantly by 16.9%-64.8% and 12.9%-61.9%, respectively; the ammonium nitrogen ($\mathrm{NH}_{4}^{+}-\mathrm{N}$) content was significantly decreased by 11.6%-24.8% in the 0-40 cm layer, yet it was increased to the 1.4 times of the CK in the>40-60 cm layer. Compared with the CK, the biological nitrogen fixation potential in 0-20 cm soil layer was significantly enhanced by 25.2% with straw returning, but the denitrification potential in 0-40 cm soil layer was weakened by 7.8%-82.0%. The copy number of functional genes of nifH, nirS, nirK and nosZ was significantly increased with straw returning than that of CK, which indicated that the growth of nitrogen fixing and denitrifying bacteria was enhanced. The results of structural equation model analysis showed that the changes of soil DOC content, soil nitrogen fixing and denitrifying bacteria abundance had direct influence on the content and distribution of soil $\mathrm{NO}_{3}^{-}-\mathrm{N}$ and $\mathrm{NH}_{4}^{+}-\mathrm{N}$ after straw returning, and the abundance of nitrogen fixing bacteria had the largest direct effect value on the content and distribution of soil $\mathrm{NO}_{3}^{-}-\mathrm{N}$ and $\mathrm{NH}_{4}^{+}-\mathrm{N}$, which was 71.9% and -60.8%, respectively.

Key words: straw returning, soil organic carbon, soil mineral nitrogen, nitrogen-fixing bacteria, denitrifying bacteria

中图分类号:

王云龙, 贾生强, 崔玲宇, 吕豪豪, 沈阿林, 苏瑶. 秸秆还田下土壤碳氮分布与固氮和反硝化细菌种群的相互影响[J]. 浙江农业学报, 2025, 37(10): 2150-2164.

WANG Yunlong, JIA Shengqiang, CUI Lingyu, LYU Haohao, SHEN Alin, SU Yao. Interactions of soil carbon and nitrogen distribution with nitrogen fixing and denitrifying bacteria community under straw returning[J]. Acta Agriculturae Zhejiangensis, 2025, 37(10): 2150-2164.

图/表 10

表1 引物信息

Table 1 Basic information of primers

目的基因 Target gene	引物 Primer	引物序列 Primer sequence (5'→3')
nifH	nifHF	AAAGGYGGWATCGGYAARTCCACCAC
	nifHR	TTGTTSGCSGCRTACATSGCCATCAT
nirK	F1aCu	ATC ATG GTSCTG CCG CG
	R3Cu	GCC TCG ATC AGR TTG TGG TT
nirS	cd3aF	GTS AAC GTS AAG GAR ACS GG
	R3cdR	GAS TTC GGR TGS GTC TTG A
nosZ	NosF2	GGG CTB GGG CCR TTG CA
	NosR2	GAA GCG RTC CTT SGA RAA CTT G

图1 不同处理的土壤有机碳和氮素组分含量 SOC,土壤有机碳含量;SMOC,土壤矿物结合态有机碳含量;SPOC,土壤颗粒有机碳含量;SDOC,土壤溶解有机碳含量;STN,土壤全氮含量;SAN,土壤碱解氮含量; SNH 4 +-N,土壤铵态氮含量; SNO 3 --N,土壤硝态氮含量。下同。

Fig.1 Soil organic carbon and nitrogen content under treatments SOC, Soil organic carbon content; SMOC, Soil mineral-incorporated organic carbon content; SPOC, Soil particulate organic carbon content; SDOC, Soil dissolved organic carbon; STN, Soil total nitrogen content; SAN, Soil alkali-hydrolyzable nitrogen content; SNH 4 +-N, Soil ammonium nitrogen content; SNO 3 --N, Soil nitrate nitrogen content. The same as below.

表2 不同处理0~60 cm土层的土壤生物固氮潜势和反硝化潜势

Table 2 Biological nitrogen fixation and denitrification potential at 0-60 cm soil layer under treatments μg·kg-1·d-1

处理 Treatment	土壤深度 Soil depth/cm	生物固氮潜势 Biological nitrogen fixation potential	生物反硝化潜势 Biological denitrification potential
CK	0~20	2.46±0.33 b	4.06±0.23 a
	>20~40	0.51±0.17 c	1.22±0.03 c
	>40~60	0.14±0.04 d	0.30±0.04 d
ST	0~20	3.08±0.03 a	3.74±0.09 b
	>20~40	0.66±0.06 c	0.22±0.01 d
	>40~60	0.15±0.09 d	0.18±0.01 d

图2 不同处理0~60 cm土层nifH (a)、nirS (b)、nirK (c)、nosZ (d)基因的丰度图中丰度均以基因拷贝数的常用对数值表示。

Fig.2 Abundance of nifH (a), nirS (b), nirK (c), nosZ (d) genes in 0-60 cm soil layer under treatments Abundance in the above figure is expressed as the common logarithm value of gene copies.

图3 nifH(a)、nirS(b)、nirK(c)、nosZ(d)基因丰度与土壤碳、氮组分的相关性 “*”和“**”分别表示显著(p<0.05)和极显著(p<0.01)相关。R2,决定系数。

Fig.3 Correlation analysis of abundance of nifH (a), nirK (b), nirS (c) and nosZ (d) genes with soil carbon and nitrogen components “*” and “**” indicate significant correlation at p<0.05 and p<0.01, respectively. R2, Coefficient of determination.

图4 土壤nifH型固氮细菌(a、b、c)和nirS型反硝化细菌(d、e、f)种群的α多样性指数 “*”表示差异显著(p<0.05)。

Fig.4 Alpha diversity index of soil nifH type nitrogen fixing bacteria (a, b, c) and nirS type denitrifying bacteria (d, e, f) “*” indicates significant difference at p<0.05.

图5 土壤nifH型固氮细菌(a)和nirS型反硝化细菌(b)种群结构差异的主坐标分析(PCoA) PCoA1,第1主坐标;PcoA2,第2主坐标。

Fig.5 Principal co-ordinates analysis (PcoA) of differences in population structure of nifH type nitrogen-fixing bacteria (a) and nirS type denitrifying bacteria (b) in soil PCoA1, Principle coordinate 1; PCoA2, Principle coordinate 2.

图6 不同处理0~20、>20~40、>40~60 cm土层存在显著差异的nifH型固氮菌属(a)和nirS型反硝化菌属(b)

Fig.6 nifH nitrogen-fixing bacteria (a) and nirS denitrifying bacteria (b) with significant differences in 0-20,>20~40,>40~60 cm soil layers under treatments

图7 土壤碳氮组分与nifH型固氮细菌(a、b)和nirS型反硝化细菌(c、d)种群结构及主要菌属的排序回归分析(a、c)和相关分析(b、d) “*”“**”和“***”分别表示在p<0.05、p<0.01和p<0.001水平上显著。unclassified_p_Proteaobacteria,未分类的变形菌门细菌;unclassified_k_norank_d_Bacteria,未分类的细菌;unclassified_c_Deltaproteobacteria,未分类的δ-变形菌纲细菌;unclassified_d_unclassified,未分类的细菌;Geobacter,地杆菌属;unclassified_c_Alphaproteobacteria,未分类的α-变形菌纲细菌;Anaeromyxobacter,厌氧菌属;unclassified_o_Rhizobiales,未分类的根瘤菌目细菌;norank_p_environmental_samples,未分类的环境样本细菌;unclassified_c_Betaproteobacteria,未分类的β-变形菌纲细菌;Rhodanobacter,罗河杆菌属;Azospira,固氮螺菌属;unclassified_f_unclassified_Burkholderiales,未分类的伯克氏菌目细菌。

Fig.7 Sequencing regression analysis (a, c) and correlation analysis (b, d) of soil cabon and nitrogen components with nifH type nitrogen-fixing bacteria (a, b) and nirS type denitrifying bacteria (c, d) population structure and main bacteria genera “*” “**” “***” indicate significant level of p<0.05, p<0.01, p<0.001, respectively.

图8 基于结构方程模型(SEM)的土壤溶解性有机碳、土壤固氮和反硝化菌群丰度及组成与土壤矿质氮含量间的路径 df,自由度;RMSEA,近似误差均方根;GFI,拟合优度;CFI,比较拟合指数。

Fig.8 Structural equation modelling (SEM) showing pathways between soil dissolved oraganic carbon (DOC) content, soil nitrogen fixation and denitrification bacterial abundance and community composition, and soil mineral nitrogen content df,Degree of freedom; RMSEA, Root mean square error of approximation; GFI, Goodness of fit; CFI, Comparative fitting index.

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秸秆还田下土壤碳氮分布与固氮和反硝化细菌种群的相互影响

Interactions of soil carbon and nitrogen distribution with nitrogen fixing and denitrifying bacteria community under straw returning

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