Effect of Organic-inorganic Combined Fertilization on Sulfur-cycling Microbial Communities in Karst Dryland Soils

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

Abstract

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

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, 2026, 42(5): 222-233.

Figures/Tables 12

References 31

[1]	Zheng CY, Li CL, Tian LB, et al. Mixture of controlled-release and normal urea to improve maize root development, post-silking plant growth, and grain filling [J]. Eur J Agron, 2023, 151: 126994.
[2]	刘传和, 贺涵, 邵雪花, 等. 不同覆盖处理对菠萝园土壤代谢物和细菌群落结构的影响 [J]. 生物技术通报, 2024, 40(7): 247-258.
	Liu CH, He H, Shao XH, et al. Analysis of differential metabolites and bacterial community structure in the soils of a pineapple orchard under different mulching treatments [J]. Biotechnol Bull, 2024, 40(7): 247-258.
[3]	Guo JH, Liu XJ, Zhang Y, et al. Significant acidification in major Chinese croplands [J]. Science, 2010, 327(5968): 1008-1010.
[4]	Xing YY, Xie YX, Wang XK. Enhancing soil health through balanced fertilization: a pathway to sustainable agriculture and food security [J]. Front Microbiol, 2025, 16: 1536524.
[5]	傅伟, 刘坤平, 陈洪松, 等. 等氮配施有机肥对喀斯特峰丛洼地农田作物产量与养分平衡的影响 [J]. 中国生态农业学报, 2017, 25(6): 812-820.
	Fu W, Liu KP, Chen HS, et al. Effect of partial replacement of inorganic N with organic manure on crop yield and soil nutrient balance in arable ecosystem in karst peak-cluster depression [J]. Chin J Eco Agric, 2017, 25(6): 812-820.
[6]	赵娜, 王小利, 何进, 等. 有机肥替代化学氮肥对黄壤活性有机碳组分、酶活性及作物产量的影响 [J]. 环境科学, 2024, 45(7): 4196-4205.
	Zhao N, Wang XL, He J, et al. Effects of replacing chemical nitrogen fertilizer with organic fertilizer on active organic carbon fractions, enzyme activities, and crop yield in yellow soil [J]. Environ Sci, 2024, 45(7): 4196-4205.
[7]	Zhang JJ, Huang JQ, Wen J, et al. Phosphorus fractions and their transformation in coupling with organic carbon cycling after seven-year manure application in subtropical soil [J]. Soil Tillage Res, 2025, 251: 106535.
[8]	路慧英, 周怀平, 杨振兴, 等. 长期氮磷化肥和有机肥配施对褐土钾素平衡及不同形态的影响 [J]. 山西农业科学, 2013, 41(1): 60-65.
	Lu HY, Zhou HP, Yang ZX, et al. Influence of long-term nitrogen, phosphorus fertilizers and organic complex fertilizations on cinnamon potassium balance and forms [J]. J Shanxi Agric Sci, 2013, 41(1): 60-65.
[9]	张伟. 西南喀斯特坡地土壤硫的生物地球化学循环研究 [D]. 贵阳: 中国科学院地球化学研究所, 2009.
	Zhang W. Study on biogeochemical cycle of soil sulfur in karst sloping land in southwest China [D]. Guiyang: Institute of Geochemistry, Chinese Academy of Sciences, 2009.
[10]	张伟, 张丽丽. 喀斯特小流域石灰土硫形态和硫酸盐还原菌分布特征 [J]. 生态与农村环境学报, 2017, 33(7): 645-652.
	Zhang W, Zhang LL. Forms of sulfur and distribution of sulfate-reducing bacteria in limestone soil of small karst catchment [J]. J Ecol Rural Environ, 2017, 33(7): 645-652.
[11]	鲍士旦. 土壤农化分析 [M]. 3版. 北京: 中国农业出版社, 2000.
	Bao SD. Soil and agricultural chemistry analysis [M]. 3rd ed. Beijing: China Agriculture Press, 2000.
[12]	Li DH, Liu CM, Luo RB, et al. MEGAHIT: an ultra-fast single-node solution for large and complex metagenomics assembly via succinct de Bruijn graph [J]. Bioinformatics, 2015, 31(10): 1674-1676.
[13]	Hyatt D, Chen GL, LoCascio PF, et al. Prodigal: prokaryotic gene recognition and translation initiation site identification [J]. BMC Bioinform, 2010, 11: 119.
[14]	Li WZ, Godzik A. Cd-hit: a fast program for clustering and comparing large sets of protein or nucleotide sequences [J]. Bioinformatics, 2006, 22(13): 1658-1659.
[15]	Kanehisa M, Goto S. KEGG Kyoto encyclopedia of genes and genomes [J]. Nucleic Acids Res, 2000, 28(1): 27-30.
[16]	Li JN, Zhao J, Liao XH, et al. Pathways of soil organic carbon accumulation are related to microbial life history strategies in fertilized agroecosystems [J]. Sci Total Environ, 2024, 927: 172191.
[17]	Zhang F, Chen MR, Xing YY, et al. Reduction of nitrogen fertilizer and simultaneously application of organic fertilizer optimizes yield, water productivity and nitrogen metabolism of spring maize by improving soil properties in the Loess Plateau of China [J]. J Agric Food Res, 2025, 19: 101634.
[18]	李青林. 喀斯特不同生境土壤理化性质差异及其空间分异特征 [D]. 贵阳: 贵州大学, 2021.
	Li QL. Differences of soil physical and chemical properties and their spatial differentiation characteristics in different karst habitats [D]. Guiyang: Guizhou University, 2021.
[19]	Geisseler D, Scow KM. Long-term effects of mineral fertilizers on soil microorganisms-A review [J]. Soil Biol Biochem, 2014, 75: 54-63.
[20]	Cui XW, Zhang YZ, Gao JS, et al. Long-term combined application of manure and chemical fertilizer sustained higher nutrient status and rhizospheric bacterial diversity in reddish paddy soil of Central South China [J]. Sci Rep, 2018, 8: 16554.
[21]	Li Y, Liu XM, Zhang L, et al. Effects of short-term application of chemical and organic fertilizers on bacterial diversity of cornfield soil in a karst area [J]. J Soil Sci Plant Nutr, 2020, 20(4): 2048-2058.
[22]	Leff JW, Jones SE, Prober SM, et al. Consistent responses of soil microbial communities to elevated nutrient inputs in grasslands across the globe [J]. Proc Natl Acad Sci USA, 2015, 112(35): 10967-10972.
[23]	Fontaine S, Mariotti A, Abbadie L. The priming effect of organic matter: a question of microbial competition? [J]. Soil Biol Biochem, 2003, 35(6): 837-843.
[24]	Niu GX, Zhong BQ, Wang RZ, et al. Effects of nitrogen and water addition on soil carbon, nitrogen, phosphorus, sulfur, and their stoichiometry along soil profile in a semi-arid steppe [J]. J Soils Sediments, 2023, 23(9): 3298-3309.
[25]	俞月凤, 韦建华, 张俊辉, 等. 喀斯特地区不同碳酸盐岩发育耕地土壤团聚体稳定性及有机碳剖面分布特征 [J]. 土壤通报, 2024, 55(5): 1292-1301.
	Yu YF, Wei JH, Zhang JH, et al. Stability of soil aggregates and profile distribution characteristics of organic carbon in farmland of different carbonate rocks [J]. Chin J Soil Sci, 2024, 55(5): 1292-1301.
[26]	陈运霜, 沈育伊, 曹杨, 等. 喀斯特湿地转变为农用地对土壤有机硫形态及芳基硫酸酯酶活性的影响 [J]. 植物营养与肥料学报, 2023, 29(12): 2232-2246.
	Chen YS, Shen YY, Cao Y, et al. Effects of natural wetland conversion to farmland on soil organic sulfur forms and arylsulphatase activities in karst areas [J]. Plant Nutr Fert Sci, 2023, 29(12): 2232-2246.
[27]	Liu HY, Dai ZC, Wang YJ, et al. Interacting effects of water and nitrogen addition on soil-plant sulfur dynamics in a semi-arid grassland [J]. Geoderma, 2024, 442: 116796.
[28]	Mitsuta A, Lourenço KS, de Oliveira BG, et al. Soil pH determines the shift of key microbial energy metabolic pathways associated with soil nutrient cycle [J]. Appl Soil Ecol, 2025, 208: 105992.
[29]	Xiao D, Xiao LM, Che RX, et al. Phosphorus but not nitrogen addition significantly changes diazotroph diversity and community composition in typical karst grassland soil [J]. Agric Ecosyst Environ, 2020, 301: 106987.
[30]	Zhang W, Zhao J, Pan FJ, et al. Changes in nitrogen and phosphorus limitation during secondary succession in a karst region in southwest China [J]. Plant Soil, 2015, 391(1/2): 77-91.
[31]	Zhang H, Dou ZX, Bi WH, et al. Multi-omics study of sulfur metabolism affecting functional microbial community succession during aerobic solid-state fermentation [J]. Bioresour Technol, 2023, 387: 129664.

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

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

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

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

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