有机-无机配施对喀斯特旱地土壤硫循环功能微生物群落的影响

doi:10.13560/j.cnki.biotech.bull.1985.2025-1451

摘要/Abstract

摘要：

目的针对喀斯特旱地土壤硫素缺乏、硫循环功能微生物群落脆弱等问题，研究有机-无机配施对硫循环功能微生物群落结构、核心功能基因及土壤理化性质的影响，为区域土壤硫循环功能维持与合理施肥提供理论依据。方法基于2009-2024年长期田间定位试验，设置CK（不施肥）、CF（全量化学施肥）、30% MF（30%农家肥+70%化肥）和60% MF（60%农家肥+ 40%化肥）4个处理，结合宏基因组测序与土壤理化分析，比较不同施肥模式下硫循环微生物群落结构、功能基因表达及其与土壤理化性质的关联特征。结果 60% MF处理显著增加土壤有机碳、全氮、碱解氮含量；30% MF处理对硫素积累最为显著。主成分分析表明，有机肥添加是驱动硫循环功能微生物群落差异的关键因素，CK与CF集中在负轴，有机-无机配施处理分布于正轴。假单胞菌门（硫氧化优势类群）和放线菌门（硫酸盐还原优势类群）为优势功能菌门；30% MF处理显著提高无机与有机硫转化关联基因（ssuA）、有机硫转化相关基因（dmdA）、硫还原相关基因（ttrA）丰度。基于门水平微生物群落结构、硫循环功能基因及代谢途径的相关性分析表明，磷素是影响喀斯特旱地硫循环功能基因变化的关键因子。结论 30%有机配施利于土壤硫素积累，60%有机配施更具快速培肥优势，应重视磷素管理以维持硫循环功能，为喀斯特旱地差异化施肥提供依据。

关键词: 有机-无机配施, 喀斯特, 硫循环, 土壤微生物, 功能基因

Abstract:

Objective To address the issues of sulfur deficiency and the vulnerability of sulfur-cycling functional microbial communities in karst dryland soils, this study investigated the effects of organic-inorganic combined fertilization on the structure of sulfur-cycling functional microbial communities, core functional genes, and soil physicochemical properties, providing a theoretical basis for maintaining the sulfur-cycling function and rational fertilization in the region. Method Based on a long-term field experiment from 2009 to 2024, four treatments were set up: CK (no fertilization), CF (full chemical fertilization), 30% MF (30% farmyard manure + 70% chemical fertilizer), and 60% MF (60% farmyard manure + 40% chemical fertilizer). By integrating metagenomic sequencing and soil physicochemical analysis, the differences in sulfur-cycling microbial community structure, expression of functional genes, and their correlations with soil physicochemical properties under different fertilization patterns were compared. Result The 60% MF treatment significantly increased soil organic carbon, total nitrogen, and available nitrogen content; the 30% MF treatment was most effective in sulfur accumulation. The principal component analysis indicated that the addition of organic fertilizer was the key factor driving the differences in sulfur-cycling functional microbial communities, with CK and CF concentrated on the negative axis and organic-inorganic combined fertilization treatments distributed on the positive axis. The Pseudomonadota phylum (dominant sulfur-oxidizing group) and Actinomycetota phylum (dominant sulfate-reducing group) were the dominant functional phyla; the 30% MF treatment significantly increased the abundance of genes related to inorganic and organic sulfur transformation (ssuA), organic sulfur transformation (dmdA), and sulfur reduction (ttrA). Correlation analysis based on microbial community structure at the phylum level, sulfur-cycling functional genes, and metabolic pathways indicated that phosphorus was a key factor influencing the changes in sulfur-cycling functional genes in karst dryland soils. Conclusion A 30% organic-inorganic combined fertilization is beneficial for soil sulfur accumulation, while a 60% organic-inorganic combined fertilization has a more rapid soil fertility improvement advantage. Phosphorus management should be emphasized to maintain the sulfur-cycling function, providing a basis for differentiated fertilization in karst dryland soils.

Key words: organic-inorganic combined application, karst, sulfur-cycling, soil microorganisms, functional gene

岳源怡, 黄世雄, 刘坤平, 冯书珍. 有机-无机配施对喀斯特旱地土壤硫循环功能微生物群落的影响[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2025-1451.

YUE Yuan-yi, HUANG Shi-xiong, LIU Kun-ping, FENG Shu-zhen. Effect of Organic-inorganic Combined Fertilization on Sulfur-cycling Microbial Communities in Karst Dryland Soils[J]. Biotechnology Bulletin, doi: 10.13560/j.cnki.biotech.bull.1985.2025-1451.

图/表 12

表1 有机-无机配施条件下喀斯特旱地土壤的理化性质

Table 1 Soil physicochemical properties in karst dryland under organic-inorganic combined fertilization

项目 Item	CK	CF	30% MF	60% MF
pH	6.72±0.02a	7.16±0.11a	6.64±0.01a	6.93±0.04a
有机碳SOC（g/kg）	21.07±0.47a	20.64±0.46a	22.81±0.55a	26.86±0.52b
全氮TN（g/kg）	2.02±0.04a	2.03±0.07a	2.18±0.02a	2.54±0.04b
全磷TP（g/kg）	1.85±0.01a	0.94±0.03b	0.58±0.01c	0.52±0.04c
全钾TK（g/kg）	3.10±0.02a	2.92±0.07a	3.10±0.07a	3.18±0.05a
全硫TS（g/kg）	0.52±0.01c	0.54±0.02bc	0.62±0.03a	0.60±0.01ab
碱解氮AN（mg/kg）	110.79±4.69a	100.62±7.08a	124.99±6.28ab	134.69±2.81b
速效磷AP（mg/kg）	23.66±1.83b	7.97±0.77c	24.52±0.14a	31.53±2.01b
速效钾AK（mg/kg）	302.63±9.36a	60.80±0.69c	265.83±2.64b	310.85±4.65a
有效硫AS（mg/kg）	5.22±0.52c	6.00±0.91bc	9.91±1.31a	8.87±0.53ab
含水率SWC（%）	3.902 ± 0.313a	3.180 ± 0.083a	3.208 ± 0.365a	2.748 ± 0.074a

表2 有机-无机配施条件下土壤基因组DNA测序数据统计

Table 2 Statistics of soil genome DNA sequencing data under organic-inorganic combined fertilization

处理 Treatment	原始数据 Raw data		清除后数据 Clean data		比例 Percentage （%）
处理 Treatment	原始序列条数 Raw reads	原始总长度（碱基对） Raw total length （bp）	清除后序列条数 Clean reads	清除后总长度（碱基对） Clean total length （bp）	比例 Percentage （%）
CK	69 368 338	10 474 619 038	68 198 170	10 259 379 377	98.31
	73 160 698	11 047 265 398	71 871 992	10 809 932 382	98.24
	71 108 096	10 737 322 496	70 004 272	10 530 770 856	98.45
	66 933 406	10 106 944 306	65 969 148	9 928 087 529	98.56
CF	74 394 464	11 233 564 064	73 193 682	11 010 958 103	98.39
	73 464 634	11 093 159 734	72 232 630	10 864 136 939	98.32
	72 022 608	10 875 413 808	70 890 576	10 656 228 389	98.43
	74 793 672	11 293 844 472	73 515 758	11 057 358 499	98.29
30% MF	73 751 614	11 136 493 714	72 530 788	10 908 722 230	98.34
	71 971 366	10 867 676 266	70 829 750	10 655 528 925	98.41
	72 361 960	10 926 655 960	71 130 528	10 697 479 080	98.30
	78 549 904	11 861 035 504	77 175 014	11 608 945 637	98.25
60% MF	69 313 388	10 466 321 588	68 258 190	10 270 223 217	98.48
	70 273 496	10 611 297 896	69 233 192	10 417 451 166	98.52
	70 990 218	10 719 522 918	69 927 030	10 521 520 756	98.50
	66 020 820	9 969 143 820	64 929 700	9 769 923 406	98.35

图1 有机-无机配施条件下喀斯特旱地土壤微生物群落 OTUs 数

Fig. 1 OTUs of soil microbial community in karst dryland under organic-inorganic combined fertilization

表3 四个施肥处理土壤中共有物种中优势门类的组成分析

Table 3 Composition analysis of dominant phyla among common species in soils under four fertilization treatments

门类 Phylum	OTU数量 Number of OTUs	相对丰度 Relative abundance （%）
Pseudomonadota	10 963	31.70
Actinomycetota	9 932	28.71
Candidatus Rokubacteria	38 39	11.10
Acidobacteriota	1 643	4.75
Chloroflexota	1 184	3.42

图2 有机-无机配施条件下喀斯特旱地土壤30% MF和60% MF处理共有的微生物物种分布

Fig. 2 Distribution of common microbial species between 30% MF and 60% MF treatments under organic-inorganic combined fertilization in karst dryland soils

图 3 有机-无机配施条件下喀斯特旱地土壤微生物群落的 PCA 分析

Fig. 3 PCA analysis of soil microbial communities in karst dryland soils under organic-inorganic fertilization

图4 有机-无机配施条件下喀斯特旱地土壤硫循环微生物群落相对丰度A：门水平；B：纲水平；C：属水平；D：种水平

Fig. 4 Relative abundance of soil sulfur-cycling microbial communities in karst dryland under organic-inorganic fertilizationA: Phylum level; B: class level; C: genus level; D: species level

图5 有机-无机配施条件下喀斯特旱地土壤硫循环功能基因的相对表达量1：硫还原；2：硫歧化反应；3：硫氧化系统；4：其他；5：有机硫转化；6：无机硫与有机硫转化的关联；7：异化硫还原与氧化；8：同化硫酸盐还原。不同小写字母表示各处理平均值之间存在显著性差异（P<0.05）

Fig. 5 Relative expression of soil sulfur-cycling functional genes in karst dryland under organic-inorganic fertilization1: Sulfur reduction; 2: sulfur disproportionation; 3: SOX systems; 4: others; 5: organic sulfur transformation; 6: correlation between inorganic and organic sulfur transformation; 7: dissimilatory sulfur reduction and oxidation; 8:assimilatory sulfate reduction. Different lowercase letters indicate significant differences among the means of each treatment (P<0.05)

图6 有机-无机配施条件下喀斯特旱地土壤硫循环功能基因的相对丰度

Fig. 6 Relative abundance of soil sulfur-cycling functional genes in karst dryland under organic-inorganic fertilization

图 7 门水平上前 10种微生物与土壤理化性质的相关分析1: Acidobacteriota; 2: Actinomycetota; 3: Bacillota; 4: Candidatus_Binatota; 5: Candidatus_Rokubacteria; 6: Candidatus_Tectomicrobia; 7: Chloroflexota; 8: Cyanobacteriota; 9: Nitrososphaerota; 10: Pseudomonadota.*P<0.05; **P<0.01; ***P<0.001. The same below

Fig. 7 Correlation analysis between the top ten microorganisms and environmental factors at the phylum level

图8 土壤硫循环功能基因与土壤理化性质间的相关分析

Fig. 8 Correlation analysis between soil sulfur-cycling functional genes and environmental factors

图9 土壤硫循环途径与土壤理化性质之间的相关分析

Fig. 9 Correlation analysis between soil sulfur-cycling pathways and environmental factors

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