Effects of Stropharia rugosoannulata Substrate on Tobacco Bacterial Wilt and Soil Microbial Function

doi:10.13560/j.cnki.biotech.bull.1985.2025-1210

Abstract

Abstract:

Objective To address the deterioration of soil physicochemical properties caused by continuous tobacco cropping and the aggravation of bacterial wilt caused by Ralstonia solanacearum, this study investigated the regulatory mechanisms of Stropharia rugosoannulata substrate (SMS) on the rhizosphere microecology and disease incidence of tobacco. Method A pot experiment was conducted using the cultivar ‘Yunyan 87’ planted in soil affected by continuous cropping. The experiment included a control group (CK) and a treatment group (BK) amended with 0.5 kg of S. rugosoannulata substrate. Forty-five days post-transplanting, the disease index was assessed, the physicochemical properties of rhizosphere soil were measured, and metagenomic sequencing was employed to analyze differences in microbial community structure and metabolic functional pathways. Result The SMS amendment significantly altered soil physicochemical properties. Compared to the CK group, the BK group demonstrated significantly higher levels of available phosphorus and organic matter (P<0.01) and nitrogen content significantly increased (P<0.05), while available potassium content significantly reduced (P<0.01). Furthermore, the occurrence of bacterial wilt was effectively inhibited (P<0.05). Microbial community analysis revealed that the Simpson diversity index and Pielou's evenness index in the rhizosphere soil of BK were significantly lower than those of the control. Metagenomic functional analysis indicated a significant upregulation of carbohydrate metabolism pathways in the BK (P<0.05). Specifically, there was a marked increase in the abundance of genes associated with fructose and mannose metabolism; cysteine and methionine metabolism; and alanine, aspartate, and glutamate metabolism. This included key genes involved in alginate synthesis (manB, alg44, and algG) and amino acid metabolism (asdA and asnA). Conclusion S. rugosoannulata substrate optimizes soil physicochemical properties by increasing organic matter, nitrogen, and phosphorus levels while reducing available potassium. It remodels the rhizosphere microecological structure by enriching beneficial microbial groups through directional selection. Concurrently, it activates key metabolic pathways involving fructose, mannose, and amino acids, which aids in inhibiting pathogen biofilm formation and enhancing plant immunity and growth, thereby significantly reducing the incidence of tobacco bacterial wilt.

Key words: bacterial wilt, mushroom substrate, rhizosphere microorganisms, metagenomics, metabolic pathway

HU Kuo-jun, HUANG Xiao-hui, HUANG Yi, ZHANG Yu-yu, DENG Zheng-yu, GUO Jun, ZENG Yin, YIN Hua-qun, ZHOU Xiang-ping, MENG De-long. Effects of Stropharia rugosoannulata Substrate on Tobacco Bacterial Wilt and Soil Microbial Function[J]. Biotechnology Bulletin, 2026, 42(5): 124-133.

Figures/Tables 7

Table 1 Soil physicochemical properties and disease index in control and S. rugosoannulata substrate-treated groups

土壤理化性质 Soil physicochemical properties	CK	BK	显著性 Significance
速效钾 AK （mg/kg）	655.00±108.70	281.25±34.73	**
速效磷 AP （mg/kg）	308.81±28.00	438.73±43.32	**
有机质 OM （mg/kg）	142.60±4.73	155.67±2.19	**
氮 N （mg/kg）	661.95±65.68	810.9±45.79	*
含水量 Water （%）	51.87±2.89	50.56±1.75
酸碱度 pH	7.15±0.19	7.43±0.1
总钾 TK （mg/kg）	13.36±0.49	14.04±0.42
总磷 TP （mg/kg）	2.44±0.18	2.4±0.14
病情指数 Disease index	61.1±9.65	30.6±2.25	*

Fig. 1 Diversity indices of the control group and S. rugosoannulata substrate treatment groupA: Simpson diversity index of the rhizosphere microbial community in the control and S. rugosoannulata substrate-treated groups. B: Pielou evenness index of the rhizosphere microbial community in the control and S. rugosoannulata substrate-treated groups. C: Bray-Curtis distance of the rhizosphere microbial community in the control and S. rugosoannulata substrate-treated groups. D: Principal coordinate analysis (PCoA) plot of the rhizosphere microbial community in the control and S. rugosoannulata substrate-treated groups. Different lowercase letters (a, b) indicate significant differences between treatments (P<0.05); the same letter indicates no significant difference. The same below

Fig. 2 Stacked bar plot of relative abundance at the genus level of rhizosphere soil microbial communities in control group and S. rugosoannulata substrate treatment group

Fig. 3 KEGG pathway abundance summary and pathway differential analysis between the control group and S. rugosoannulata substrate treatment groupA: KEGG Level 1 pathway RPKM summary of control group and S. rugosoannulata substrate treatment group. B: KEGG Level 2 pathway RPKM summary and differences of control group and S. rugosoannulata substrate treatment group. C: KEGG Level 3 pathway RPKM summary and differences of control group and S. rugosoannulata substrate treatment group

Fig. 4 Community functional diversity of control group and S. rugosoannulata substrate treatment groupA: Community functional Simpson diversity index of the control group and S. rugosoannulata substrate treatment group. B: Community functional Bray-Curtis distance of the control group and S. rugosoannulata substrate treatment group. C: Community functional principal coordinate analysis (PCoA) plot of the control group and S. rugosoannulata substrate treatment group

Fig. 5 Fructose and mannose metabolism pathway (A), cysteine and methionine metabolism pathway (B), and alanine, aspartate and glutamate metabolism pathway (C)

Fig. 6 Mantel test and Pearson correlation heatmapDI: Disease index; Ralstonia: Ralstonia solanacearum; Ko00051: fructose and mannose metabolism pathway; Ko00250: cysteine and methionine metabolism pathway; Ko00270: alanine, aspartate, and glutamate metabolism pathway

References 40

[1]	Yang BY, Feng CC, Jiang H, et al. Effects of long-term continuous cropping on microbial community structure and function in tobacco rhizosphere soil [J]. Front Microbiol, 2025, 16: 1496385.
[2]	Chen YD, Yang L, Zhang LM, et al. Autotoxins in continuous tobacco cropping soils and their management [J]. Front Plant Sci, 2023, 14: 1106033.
[3]	Gu KY, Liu XL, Liu M, et al. Tobacco intercropping enhances soil fertility by improving synergic interactions between soil physicochemical and microbial properties [J]. Front Microbiol, 2025, 16: 1647493.
[4]	Xia H, Jiang CQ, Riaz M, et al. Impacts of continuous cropping on soil fertility, microbial communities, and crop growth under different tobacco varieties in a field study [J]. Environ Sci Eur, 2025, 37(1): 5.
[5]	Gong B, He Y, Luo ZB, et al. Response of rhizosphere soil physicochemical properties and microbial community structure to continuous cultivation of tobacco [J]. Ann Microbiol, 2024, 74(1): 4.
[6]	Li GT, Gong PF, Zhou J, et al. The succession of rhizosphere microbial community in the continuous cropping soil of tobacco [J]. Front Environ Sci, 2024, 11: 1251938.
[7]	Ding MJ, Dai HX, He Y, et al. Continuous cropping system altered soil microbial communities and nutrient cycles [J]. Front Microbiol, 2024, 15: 1374550.
[8]	Zhang ZH, Huang WX, Jia W, et al. Combined use of green fertilizer and soil conditioner improves soil physicochemical properties, microbial community structure and tobacco quality in continuous cropping [J]. Ann Microbiol, 2025, 75(1): 14.
[9]	Feng JN, Chen LL, Xia TY, et al. Microbial fertilizer regulates C: N: P stoichiometry and alleviates phosphorus limitation in flue-cured tobacco planting soil [J]. Sci Rep, 2023, 13: 10276.
[10]	Jiang GF, Wei Z, Xu J, et al. Bacterial wilt in China: history, current status, and future perspectives [J]. Front Plant Sci, 2017, 8: 1549.
[11]	Li YY, Feng J, Liu HL, et al. Genetic diversity and pathogenicity of Ralstonia solanacearum causing tobacco bacterial wilt in China [J]. Plant Dis, 2016, 100(7): 1288-1296.
[12]	Wang ZJ, Luo WB, Cheng SJ, et al. Ralstonia solanacearum-A soil borne hidden enemy of plants: Research development in management strategies, their action mechanism and challenges [J]. Front Plant Sci, 2023, 14: 1141902.
[13]	Wang ZB, Zhang YZ, Bo GD, et al. Ralstonia solanacearum infection disturbed the microbiome structure throughout the whole tobacco crop niche as well as the nitrogen metabolism in soil [J]. Front Bioeng Biotechnol, 2022, 10: 903555.
[14]	Yao T, Wang CY, Ren Q, et al. Bacterial wilt alters the microbial community characteristics of tobacco root and rhizosphere soil [J]. Rhizosphere, 2024, 32: 100995.
[15]	苏文英, 纪伟, 刘晓梅, 等. 连云港地区秸秆-大球盖菇-大豆-豆丹种养循环模式 [J]. 农业与技术, 2022, 42(22): 53-56.
	Su WY, Ji W, Liu XM, et al. Cyclic model of straw-Stropharia rugoso-soybean-Doudan in Lianyungang area [J]. Agric & Technol, 2022, 42(22): 53-56.
[16]	杨晓波, 郑光耀, 徐丽红, 等. 不同栽培料栽培大球盖菇比较试验 [J]. 食用菌, 2021, 43(2): 32-33, 9.
	Yang XB, Zheng GY, Xu LH, et al. Comparative test on cultivating Stropharia rugosoannulata with different cultivation materials [J]. Edible Fungi, 2021, 43(2): 32-33, 9.
[17]	He Y, Gong B, Liang TB, et al. Effects of reductive soil disinfestation on microbiological and physicochemical properties of continuous cropping soils in karst areas of Guizhou Province [J]. Ann Microbiol, 2025, 75(1): 6.
[18]	Shi HQ, Xu PW, Wu SX, et al. Analysis of rhizosphere bacterial communities of tobacco resistant and non-resistant to bacterial wilt in different regions [J]. Sci Rep, 2022, 12: 18309.
[19]	Wang XJ, Li ZH, Jin HG, et al. Effects of mushroom-tobacco rotation on microbial community structure in continuous cropping tobacco soil [J]. J Appl Microbiol, 2023: lxad088.
[20]	国家质量监督检验检疫总局中国国家标准化管理委员会. 烟草病虫害分级及调查方法: [S]. 北京: 中国标准出版社, 2009.
	Standardization Administration of the People's Republic of China. Grade and investigation method of tobacco diseases and insect pests: [S]. Beijing: Standards Press of China, 2009.
[21]	Chen SF, Zhou YQ, Chen YR, et al. Fastp: an ultra-fast all-in-one FASTQ preprocessor [J]. Bioinformatics, 2018, 34(17): i884-i890.
[22]	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.
[23]	Seemann T. Prokka: rapid prokaryotic genome annotation [J]. Bioinformatics, 2014, 30(14): 2068-2069.
[24]	Steinegger M, Söding J. MMseqs2 enables sensitive protein sequence searching for the analysis of massive data sets [J]. Nat Biotechnol, 2017, 35(11): 1026-1028.
[25]	Aroney STN, Newell RJP, Nissen JN, et al. CoverM: read alignment statistics for metagenomics [J]. Bioinformatics, 2025, 41(4): btaf147.
[26]	Cao YF, Shen ZZ, Zhang N, et al. Phosphorus availability influences disease-suppressive soil microbiome through plant-microbe interactions [J]. Microbiome, 2024, 12(1): 185.
[27]	Wu K, Yuan SF, Wang LL, et al. Effects of bio-organic fertilizer plus soil amendment on the control of tobacco bacterial wilt and composition of soil bacterial communities [J]. Biol Fertil Soils, 2014, 50(6): 961-971.
[28]	Wang S, Wang L, Li SC, et al. The win-win effects of an invasive plant biochar on a soil-crop system: controlling a bacterial soilborne disease and stabilizing the soil microbial community network [J]. Microorganisms, 2024, 12(3): 447.
[29]	Yang BY, Zhang CQ, Guan CW, et al. Analysis of the composition and function of rhizosphere microbial communities in plants with tobacco bacterial wilt disease and healthy plants [J]. Microbiol Spectr, 2024, 12(12): e0055924.
[30]	Suresh P, Shanmugaiah V, Rajagopal R, et al. Pseudomonas fluorescens VSMKU3054 mediated induced systemic resistance in tomato against Ralstonia solanacearum [J]. Physiol Mol Plant Pathol, 2022, 119: 101836.
[31]	Jiao WL, Wen J, Li N, et al. The biocontrol potentials of rhizospheric bacterium Bacillus velezensis K0T24 against mulberry bacterial wilt disease [J]. Arch Microbiol, 2024, 206(5): 213.
[32]	Wang NQ, Ping L, Mei XL, et al. Succinic acid reduces tomato bacterial wilt disease by recruiting Sphingomonas sp. [J]. Environ Microbiome, 2025, 20(1): 85.
[33]	Posas MB, Toyota K, Islam TM. Inhibition of bacterial wilt of tomato caused by Ralstonia solanacearum by sugars and amino acids [J]. Microb Environ, 2007, 22(3): 290-296.
[34]	Chen Z, Lin EQ, Lin X, et al. Metabolomic profiling of tomato root exudates induced by Ralstonia solanacearum strains of different pathogenicity: screening for metabolites conferring bacterial wilt resistance [J]. J Microbiol Biotechnol, 2025, 35: e2501033.
[35]	Wen T, Xie PH, Liu HW, et al. Tapping the rhizosphere metabolites for the prebiotic control of soil-borne bacterial wilt disease [J]. Nat Commun, 2023, 14: 4497.
[36]	Asimakis E, Shehata AA, Eisenreich W, et al. Algae and their metabolites as potential bio-pesticides [J]. Microorganisms, 2022, 10(2): 307.
[37]	Yang L, Wei ZL, Valls M, et al. Metabolic profiling of resistant and susceptible tobaccos response incited by Ralstonia pseudosolanacearum causing bacterial wilt [J]. Front Plant Sci, 2022, 12: 780429.
[38]	Murti RH, Afifah EN, Nuringtyas TR. Metabolomic response of tomatoes (Solanum lycopersicum L.) against Bacterial Wilt (Ralstonia solanacearum) using ¹H-NMR spectroscopy [J]. Plants, 2021, 10(6): 1143.
[39]	Yang XH, Huang J, Yang HX, et al. Transcriptome and metabolome profiling in different stages of infestation of Eucalyptus urophylla clones by Ralstonia solanacearum [J]. Mol Genet Genom, 2022, 297(4): 1081-1100.
[40]	Niu LL, Hu W, Wang FZ, et al. Integrated transcriptomic and metabolomic analyses elucidate the mechanism by which grafting impacts potassium utilization efficiency in tobacco [J]. BMC Plant Biol, 2025, 25(1): 94.