施氮对不同轮作模式下油菜和小麦根际土壤中有机氮矿化功能基因的影响

doi:10.3864/j.issn.0578-1752.2025.19.009

摘要/Abstract

摘要：

【目的】土壤氮素转化受微生物活动的调控，并受作物类型和施肥措施的调节。阐明稻油和稻麦轮作模式下施氮对冬季作物根际土壤有机氮矿化作用功能基因丰度的影响，加深对不同作物制度和氮肥投入下根际土壤有机氮矿化过程的理解。【方法】利用定位试验，在第7年采集稻油和稻麦两种轮作模式中不施氮和施氮处理下油菜和小麦根际土壤，分析根际有机质组分和氮的有效性的差异，并利用宏基因组测序分析根际土壤有机氮矿化功能基因的丰度。【结果】与不施氮相比，施氮显著提高了根际土壤中潜在矿化氮（PMN）、有机碳（SOC）、全氮（TN）、溶解性有机碳/氮（DOC、DON）、颗粒态有机碳/氮（POC、PON）和矿物结合态有机碳/氮（MAOC、MAON）的含量。相同施肥处理下，稻油和稻麦两种轮作模式下油菜和小麦根际土壤中有机质组分和氮的有效性之间无显著差异。与不施氮相比，施氮降低了根际土壤中编码蛋白酶（K14645）、几丁质酶（K01183）、脲酶（K01429和K01430）和精氨酸酶（K01476）功能基因的丰度，但提高了编码谷氨酸脱氢酶（K00260）功能基因的丰度。施氮处理，稻油轮作模式的油菜根际土壤编码蛋白酶（K14645）和精氨酸酶（K01476）功能基因的丰度显著高于稻麦轮作模式的小麦根际土壤。参与有机氮矿化功能基因微生物以变形菌门（Proteobacteria）、酸杆菌门（Acidobacteria）和绿弯菌门（Chloroflexi）为主。施氮显著影响根际土壤编码蛋白酶功能基因的微生物群落结构，稻油和稻麦两种模式下油菜和小麦根际土壤参与有机氮矿化功能基因的微生物群落结构无显著差异。根际土壤中PMN和SOC是影响根际土壤有机氮矿化功能基因丰度和微生物群落结构的主要驱动因子。编码蛋白酶和精氨酸酶的功能基因丰度与PMN和SOC呈显著负相关。【结论】不同作物制度和氮肥投入都能够改变根际土壤中参与有机氮矿化功能基因的丰度，潜在矿化氮（PMN）和有机碳（SOC）是影响根际土壤中有机氮矿化功能基因丰度的主要因子。

关键词: 施氮, 稻油轮作, 稻麦轮作, 根际土壤, 有机质组分, 矿化功能基因, 宏基因组

Abstract:

【Objective】Soil nitrogen transformation was affected by microbial activities and modulated by crop types and fertilization practices. Understanding the effect of nitrogen application on the abundance of functional genes involved in organic nitrogen mineralization in the rhizosphere of winter crops under rice-oilseed and rice-wheat rotations, enhanced our understanding of the organic nitrogen mineralization process across different cropping systems and nitrogen fertilization regimes.【Method】A localized field experiment was conducted to collect rhizosphere soils from rapeseed and wheat in the seventh year of rice-rapeseed and rice-wheat rotation systems, under both nitrogen application and no nitrogen treatments. This analysis focused on differences in organic matter fractions and nitrogen availability. Furthermore, metagenomic sequencing was employed to analyze the abundance of functional genes mediating organic nitrogen mineralization in the rhizosphere soil.【Result】The results indicate that, compared to the no nitrogen treatment, nitrogen addition significantly increased the concentrations of potential mineralizable nitrogen (PMN), soil organic carbon (SOC), total nitrogen (TN), dissolved organic carbon/nitrogen (DOC, DON), particulate organic carbon/nitrogen (POC, PON), and mineral-associated organic carbon/nitrogen (MAOC, MAON) in rhizosphere soil. Under identical fertilization treatments, no significant differences were observed in the organic matter fractions and nitrogen availability between the rapeseed and wheat rhizosphere soils in the rice-oilseed and rice-wheat rotation systems. Compared to the no nitrogen treatment, nitrogen application reduced the abundance of functional genes encoding protease (K14645), chitinase (K01183), urease (K01429， K01430), and arginase (K01476), while increasing the abundance of functional genes encoding glutamate dehydrogenase (K00260). In the nitrogen application treatment, the abundance of functional genes encoding protease and arginase in rapeseed rhizosphere soil within the rice-oilseed rotation system was significantly higher than that in wheat rhizosphere soil within the rice-wheat rotation system. Microorganisms involved in organic nitrogen mineralization functional genes predominantly belonged to the phyla Proteobacteria, Acidobacteria, and Chloroflexi. Nitrogen addition significantly influenced the microbial community structure of protease-encoding functional genes in rhizosphere soil, whereas no significant differences were observed in the microbial community structure of organic nitrogen mineralization functional genes between rapeseed and wheat rhizosphere soils in the rice-oilseed and rice-wheat rotation systems. PMN and SOC in rhizosphere soil were identified as the primary drivers influencing the abundance of organic nitrogen mineralization functional genes and the structure of the microbial community. The abundance of functional genes encoding protease and arginase exhibited a significant negative correlation with PMN and SOC.【Conclusion】The results of this study demonstrate that both crop species and nitrogen application under different crop rotation systems can significantly influence the abundance of functional genes involved in organic nitrogen mineralization in rhizosphere soil, while also identifying the primary factors driving the abundance of these genes in rhizosphere soil.

Key words: nitrogen application, rice-rapeseed rotation, rice-wheat rotation, rhizosphere soil, organic matter components, mineralization functional genes, metagenomics

赵剑, 任涛, 方娅婷, 杨昕, 盛倩男, 李小坤, 朱俊, 鲁剑巍. 施氮对不同轮作模式下油菜和小麦根际土壤中有机氮矿化功能基因的影响[J]. 中国农业科学, 2025, 58(19): 3919-3931.

ZHAO Jian, REN Tao, FANG YaTing, YANG Xin, SHENG QianNan, LI XiaoKun, ZHU Jun, LU JianWei. Effect of Nitrogen Application on Organic Nitrogen Mineralization Functional Genes in Rapeseed and Wheat Rhizosphere Soils Under Different Rotation Patterns[J]. Scientia Agricultura Sinica, 2025, 58(19): 3919-3931.

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链接本文: https://www.chinaagrisci.com/CN/10.3864/j.issn.0578-1752.2025.19.009

https://www.chinaagrisci.com/CN/Y2025/V58/I19/3919

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功能分组 Functional groups	KEGG (KO)	酶编号 Enzyme number	KO名称KO name
蛋白酶基因 Protease genes	K01342	EC:3.4.21.62	aprE; subtilisin
蛋白酶基因 Protease genes	K14645	EC:3.4.21.-	K14645; serine protease
肽酶基因 Peptidase genes	K01255	EC:3.4.11.1	CARP, pepA; leucyl aminopeptidase
肽酶基因 Peptidase genes	K01256	EC:3.4.11.2	pepN; aminopeptidase N
几丁质酶基因 Chitinase genes	K01183	EC:3.2.1.14	E3.2.1.14; chitinase
几丁质酶基因 Chitinase genes	K01207	EC:3.2.1.52	nagZ; beta-N-acetylhexosaminidase
脲酶基因 Urease genes	K01428	EC:3.5.1.5	ureC; urease subunit alpha
	K01429	EC:3.5.1.5	ureB; urease subunit beta
	K01430	EC:3.5.1.5	ureA; urease subunit gamma
	K14048	EC:3.5.1.5	ureAB; urease subunit gamma/beta
精氨酸酶基因 Arginase gene	K01476	EC:3.5.3.1	E3.5.3.1, rocF, arg; arginase
谷氨酸脱氢酶基因 Glutamic dehydrogenase genes	K00260	EC:1.4.1.2	gudB, rocG; glutamate dehydrogenase
	K00261	EC:1.4.1.3	GLUD1_2, gdhA; glutamate dehydrogenase (NAD(P)+)
	K00262	EC:1.4.1.4	E1.4.1.4, gdhA; glutamate dehydrogenase (NADP+)

处理 Treatment	油菜Rapeseed		小麦Wheat		Two-way ANOVA (P value)
处理 Treatment	不施氮 -N	施氮 +N	不施氮 -N	施氮 +N	作物 Crop	氮肥 Nitrogen	作物×氮肥 Crop×Nitrogen
潜在矿化氮PMN (mg∙kg^-1)	53.12±4.74b	73.07±8.01a	46.17±5.31b	78.70±5.12a	0.852	0.000	0.104
无机氮SIN (mg∙kg^-1)	10.02±0.38b	10.61±0.88a	9.59±0.80b	10.97±0.50a	0.638	0.015	0.149
有机碳SOC (g∙kg^-1)	13.35±0.56b	15.74±0.74a	13.04±0.77b	15.67±0.94a	0.676	0.000	0.807
全氮TN (g∙kg^-1)	1.36±0.11b	1.57±0.07a	1.27±0.11b	1.51±0.13a	0.275	0.007	0.837
溶解性有机碳 DOM-C (mg∙kg^-1)	60.95±6.93b	75.70±8.92a	59.57±4.31b	75.87±6.74a	0.884	0.005	0.851
溶解性有机氮 DOM-N (mg∙kg^-1)	19.26±3.25c	25.07±1.62ab	21.25±2.96bc	26.71±2.63a	0.277	0.007	0.912
颗粒态有机碳POM-C (g∙kg^-1)	3.95±0.38b	5.26±0.6a	4.13±0.43b	5.29±0.72a	0.746	0.005	0.816
颗粒态有机氮POM-N (g∙kg^-1)	0.30±0.04b	0.37±0.05a	0.31±0.04b	0.36±0.06a	0.862	0.048	0.773
矿物结合态有机碳MAOM-C (g∙kg^-1)	9.41±0.35b	10.48±0.54a	8.91±0.50b	10.38±0.64a	0.343	0.003	0.540
矿物结合态有机氮MAOM-N (g∙kg^-1)	1.07±0.08b	1.20±0.03a	0.97±0.07b	1.15±0.07a	0.082	0.003	0.561

处理 Treatment	KO	油菜Rapeseed		小麦Wheat		Two-way ANOVA (P value)
处理 Treatment	KO	不施氮 -N	施氮 +N	不施氮 -N	施氮 +N	作物 Crop	氮肥 Nitrogen	作物×氮肥 Crop×Nitrogen
蛋白酶基因 Protease genes	K01342	21.17±2.72a	22.90±1.28a	22.00±0.36a	17.90±1.51b	0.066	0.261	0.017
蛋白酶基因 Protease genes	K14645	61.90±3.53a	59.07±2.40a	61.70±1.71a	49.17±3.01b	0.013	0.001	0.016
肽酶基因 Peptidase genes	K01255	136.42±18.1a	122.67±5.52a	120.62±3.59a	115.59±14.29a	0.137	0.212	0.547
肽酶基因 Peptidase genes	K01256	44.27±10.08a	37.10±1.57a	36.67±1.27a	34.30±3.96a	0.141	0.172	0.472
几丁质酶基因 Chitinase genes	K01183	38.70±5.74a	31.77±1.53b	34.33±0.95ab	32.43±0.65b	0.321	0.035	0.188
几丁质酶基因 Chitinase genes	K01207	138.09±7.22a	127.23±4.22ab	127.41±5.13ab	117.93±12.59b	0.062	0.058	0.886
脲酶基因 Urease genes	K01428	28.17±2.02a	26.73±0.32a	28.93±0.59a	27.93±2.01a	0.278	0.188	0.804
	K01429	25.40±2.16a	22.20±1.61b	23.77±1.10ab	22.43±0.95ab	0.451	0.033	0.322
	K01430	32.57±4.51a	25.90±2.00b	31.17±2.50ab	29.53±1.44ab	0.517	0.036	0.166
	K14048	14.53±1.19a	14.03±1.00a	14.73±0.60a	13.20±1.15a	0.604	0.121	0.404
精氨酸酶基因 Arginase gene	K01476	109.16±3.52a	99.42±3.60ab	104.10±4.20a	90.13±8.19b	0.045	0.004	0.505
谷氨酸脱氢酶基因 Glutamic dehydrogenase genes	K00260	22.30±5.00b	30.87±2.99a	24.87±2.42ab	26.13±3.43ab	0.615	0.045	0.116
	K00261	111.93±16.51a	117.48±4.61a	116.57±4.13a	101.96±8.49a	0.364	0.446	0.112
	K00262	18.07±2.97a	21.90±2.19a	18.57±0.98a	21.13±4.27a	0.938	0.089	0.712