生物炭施用对大豆轮连作系统土壤固氮微生物nifH基因丰度及群落组成的影响

doi:10.3864/j.issn.0578-1752.2026.06.010

中国农业科学 ›› 2026, Vol. 59 ›› Issue (6): 1272-1285.doi: 10.3864/j.issn.0578-1752.2026.06.010

• 土壤肥料·节水灌溉·农业生态环境 • 上一篇下一篇

生物炭施用对大豆轮连作系统土壤固氮微生物nifH基因丰度及群落组成的影响

李永娟¹^,²(), 张悦彤¹^,², 王艺博¹^,², 赵长江¹^,²^,³, 宋洁¹^,³, 陈雪丽⁴, 姚钦¹^,²^,³()

¹ 黑龙江八一农垦大学农学院，黑龙江大庆 163319
² 黑龙江省秸秆资源化利用工程技术研究中心，黑龙江大庆 163319
³ 农业农村部东北平原农业绿色低碳重点实验室，黑龙江大庆 163319
⁴ 黑龙江省黑土保护利用研究院，哈尔滨 150028

收稿日期:2025-05-02 接受日期:2025-07-22 出版日期:2026-03-16 发布日期:2026-03-24
通信作者:
姚钦，E-mail：yaoqin_byau@163.com
联系方式: 李永娟，E-mail：leeyongjuanj@163.com。
基金资助:
黑龙江省自然科学基金联合引导项目(LH2023C075); 黑龙江八一农垦大学引进人才科研启动计划(XYB202005); 黑龙江省生态环境保护科研项目(HST2024TR011); 黑龙江八一农垦大学研究生创新科研项目(NXYCX2024-Y14)

Effects of Biochar Application on the Abundance and Community Composition of Nitrogen-Fixing Microbial nifH Gene in Soybean Rotation and Continuous Cropping Systems

LI YongJuan¹^,²(), ZHANG YueTong¹^,², WANG YiBo¹^,², ZHAO ChangJiang¹^,²^,³, SONG Jie¹^,³, CHEN XueLi⁴, YAO Qin¹^,²^,³()

¹ College of Agriculture, Heilongjiang Bayi Agricultural University, Daqing 163319, Heilongjiang
² Heilongjiang Engineering Research Center of Straw Resource Utilization, Daqing 163319, Heilongjiang
³ Key Laboratory of Low-Carbon Green Agriculture in Northeastern China, Ministry of Agriculture and Rural Affairs, Daqing 163319, Heilongjiang
⁴ Heilongjiang Institute of Black Soil Protection and Utilization, Harbin 150028

Received:2025-05-02 Accepted:2025-07-22 Published:2026-03-16 Online:2026-03-24

摘要/Abstract

摘要：

【目的】分析连作与轮作条件下生物炭对土壤固氮微生物功能基因群落结构及多样性的影响，为东北黑土区退化土壤的可持续改良和不同耕作制度下生物炭的精准施用提供理论依据。【方法】基于生物炭施用长期定位试验，采集连作与轮作条件下大豆成熟期生物炭施用量分别为0（B0）、5 t·hm^-2（B5）、15 t·hm^-2（B15）和25 t·hm^-2（B25）处理的土壤样品，采用荧光定量PCR方法和Illumina Miseq高通量测序技术分析土壤固氮微生物nifH基因绝对丰度和群落结构多样性。【结果】连作、轮作条件下生物炭施用均提高了土壤pH、全氮（TN）、全磷（TP）、碱解氮（AN）、速效磷（AP）和速效钾（AK）含量，但降低了土壤全钾（TK）含量。生物炭施用显著影响了固氮微生物nifH基因绝对丰度，连作和轮作条件下高量生物炭B25处理的固氮微生物nifH基因丰度比B0处理分别提升了40.3%和149.6%，且轮作条件下整体丰度显著高于连作条件，主要受全量氮、磷、钾（TN、TP、TK）及速效养分（AN、AP、AK）的综合调控。另外，连作和轮作条件下，施加适量生物炭（B15）可以显著提高固氮微生物nifH基因群落丰富度和多样性。冗余分析表明，生物炭可通过影响土壤化学性质间接驱动土壤固氮微生物nifH基因群落结构变化。固氮微生物nifH基因群落组成中的优势菌纲为α-变形菌纲（Alphaproteobacteria），其中优势菌属为慢生根瘤菌属（Bradyrhizobium），其相对丰度随着生物炭施用量的增加而上升。连作条件下B15处理慢生根瘤菌属（Bradyrhizobium）相对丰度最高，轮作条件下B25处理相对丰度最高，但与理化性质无显著相关性。而第二优势菌属斯克尔曼氏菌属（Skermanella）与碱解氮和速效磷含量呈极显著正相关，与全氮含量显著正相关。【结论】生物炭通过调控土壤养分含量优化固氮微生物nifH基因群落多样性，改善群落结构和组成，提升连作、轮作系统养分利用效率，实现土壤生态系统的良性循环。

关键词: 生物炭, 固氮微生物, nifH基因, 土壤养分, 群落结构, 大豆, 玉米, 连作, 轮作

Abstract:

【Objective】This study aimed to analyze the effects of biochar on the community structure and diversity of nitrogen-fixing microorganisms (nifH gene) in soil under continuous cropping and crop rotation, and would provide a theoretical basis for the sustainable improvement of degraded soils in the black soil of Northeast China and the precise application of biochar under different cropping systems. 【Method】 This study collected soil samples under continuous and rotational cropping conditions with biochar application rates of 0 (B0), 5 t·hm^-2 (B5), 15 t·hm^-2 (B15), and 25 t·hm^-2 (B25) at the soybean maturity stage based on a long-term biochar application field experiment. Real-time PCR and Illumina Miseq high-throughput sequencing were used to analyze the absolute abundance and community structure diversity of the nifH gene of nitrogen-fixing microorganisms. 【Result】Biochar application under both continuous cropping and crop rotation increased soil pH, total nitrogen (TN), total phosphorus (TP), alkaline hydrolyzable nitrogen (AN), available phosphorus (AP), and available potassium (AK), but decreased total potassium (TK) content. Biochar significantly affected the absolute abundance of the nifH gene. Under continuous cropping and crop rotation, the nifH gene abundance under the high-dose biochar treatment (B25) increased by 40.3% and 149.6%, respectively, compared with the control (B0). Moreover, the nifH gene abundance under crop rotation was significantly higher than under continuous cropping, regulated by the combined effects of total nutrients (TN, TP, and TK) and available nutrients (AN, AP, and AK). Additionally, moderate biochar application (B15) significantly enhanced the richness and diversity of the nifH gene community under both cropping systems. Redundancy analysis indicated that biochar indirectly drove changes in the nifH gene community structure by altering soil chemical properties. The dominant bacterial class in the nifH gene community composition of nitrogen-fixing microorganisms was Alphaproteobacteria, with Bradyrhizobium being the dominant genus, and its relative abundance increased with the increase of biochar application rates. Under continuous cropping conditions, the Bradyrhizobium genus showed the highest relative abundance under the B15 treatment, while in crop rotation condition, the highest relative abundance occurred under the B25 treatment. However, no significant correlation was found between its abundance and soil physicochemical properties. The second dominant genus, Skermanella, exhibited a highly significant positive correlation with AN and AP contents, and a significant positive correlation with TN content. 【Conclusion】 Biochar optimized the diversity of the nifH gene community in nitrogen-fixing microorganisms by regulating soil nutrient content, improved community structure and composition of the nifH gene, enhanced nutrient use efficiency under continuous cropping and rotation systems, and then finally promoted a virtuous cycle in the soil ecosystem.

Key words: biochar, nitrogen-fixing microorganisms, nifH gene, soil nutrient, soybean, maize, community composition, continuous cropping, crop rotation

李永娟, 张悦彤, 王艺博, 赵长江, 宋洁, 陈雪丽, 姚钦. 生物炭施用对大豆轮连作系统土壤固氮微生物nifH基因丰度及群落组成的影响[J]. 中国农业科学, 2026, 59(6): 1272-1285.

LI YongJuan, ZHANG YueTong, WANG YiBo, ZHAO ChangJiang, SONG Jie, CHEN XueLi, YAO Qin. Effects of Biochar Application on the Abundance and Community Composition of Nitrogen-Fixing Microbial nifH Gene in Soybean Rotation and Continuous Cropping Systems[J]. Scientia Agricultura Sinica, 2026, 59(6): 1272-1285.

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处理 Treatment		总有机氮 TON (g·kg^-1)	全氮 TN (g·kg^-1)	碱解氮 AN (mg·kg^-1)	硝态氮 NO₃^--N (mg·kg^-1)	铵态氮 NH₄⁺-N (mg·kg^-1)
连作 Continuous cropping	B0	0.61±0.02b	1.45±0.04d	85.81±0.53c	2.08±0.01a	92.63±1.00a
	B5	0.63±0.02a	1.56±0.02c	86.39±2.11ab	2.12±0.37a	81.59±0.84c
	B15	0.61±0.64a	1.62±0.03b	90.59±1.99b	1.97±0.32a	82.67±2.46c
	B25	0.68±0.02a	1.71±0.02a	99.46±3.86a	2.06±0.23a	88.50±0.91b
轮作 Crop rotation	B0	0.65±0.02ab	1.82±0.00a	118.48±2.78b	2.00±0.18c	78.92±1.40bc
	B5	0.63±0.01b	1.73±0.01b	121.28±2.78ab	4.45±0.24a	74.82±4.17c
	B15	0.69±0.00a	1.83±0.03a	122.21±0.53ab	4.42±0.29a	88.45±0.36a
	B25	0.65±0.05ab	1.79±0.04a	123.61±1.07a	2.91±0.27b	82.74±1.51b

试验处理 Treatment		pH	含水量 W (%)	总有机碳 TOC (g·kg^-1)	全磷 TP (g·kg^-1)	全钾 TK (g·kg^-1)	速效磷 AP (mg·kg^-1)	速效钾 AK (mg·kg^-1)
连作 Continuous cropping	B0	8.44±0.01a	16.50±0.05a	1.86±0.10ab	0.67±0.02d	19.21±0.26a	7.45±0.31d	304.00±2.65c
	B5	8.45±0.01a	16.53±0.05a	1.78±0.04b	0.99±0.04a	17.48±0.41b	9.38±0.12c	285.00±1.73d
	B15	8.37±0.00b	16.66±0.01a	1.81±0.04b	0.82±0.02c	16.34±0.42c	14.57±0.35b	356.67±3.21b
	B25	8.43±0.01a	16.28±0.07b	1.94±0.03a	0.87±0.03b	16.80±0.43bc	18.19±1.00a	401.67±4.73a
轮作 Crop rotation	B0	8.38±0.01b	16.32±0.02c	2.02±0.04b	0.87±0.05b	17.90±0.37a	22.62±0.39c	344.67±2.08c
	B5	8.40±0.01b	16.99±0.01b	2.07±0.03b	0.84±0.03b	15.20±0.45c	15.44±0.22d	327.00±3.46d
	B15	8.41±0.02b	16.63±0.30bc	2.28±0.07a	1.05±0.05a	16.14±0.15b	35.75±1.16b	357.67±4.04b
	B25	8.43±0.01a	17.47±0.11a	2.02±0.06b	1.06±0.03a	15.68±0.12bc	41.05±1.76a	397.67±3.51a

参数 Parameter	处理 Treatment
参数 Parameter	B0	B5	B15	B25
节点Node	208	260	219	222
边数Edge	1470	2041	2357	2157
模块化Modularity	0.56	0.53	0.45	0.44
图密度Density	0.07	0.06	0.10	0.09
平均度Average degree	14.14	15.70	21.53	19.43
平均路径长度Average path length	3.96	4.25	3.26	3.76
平均聚类系数Average clustering coefficient	0.54	0.53	0.55	0.58

生物炭施用对大豆轮连作系统土壤固氮微生物nifH基因丰度及群落组成的影响

Effects of Biochar Application on the Abundance and Community Composition of Nitrogen-Fixing Microbial nifH Gene in Soybean Rotation and Continuous Cropping Systems

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