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

Abstract

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

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

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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