大球盖菇基质对烟草青枯病和土壤微生物功能的影响

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

摘要/Abstract

摘要：

目的针对烟草连作导致的土壤理化性质恶化及青枯雷尔氏菌（Ralstonia solanacearum）引起的青枯病加剧问题，探讨大球盖菇（Stropharia rugosoannulata）基质对烟草根际土壤微生态及病害发生的调控机制。方法采用盆栽试验，以云烟87为供试品种，利用连作患病土壤设置对照组（CK）与添加0.5 kg大球盖菇基质的处理组（BK），移栽45 d后调查烟草病情指数，测定根际土壤理化性质，并利用宏基因组测序技术，分析微生物群落结构与其代谢功能通路的差异。结果大球盖菇基质处理显著改变了土壤理化性质，与对照组相比，处理组土壤速效磷、有机质含量显著提高（P<0.01），氮含量显著增加（P<0.05），而速效钾含量显著降低（P<0.01），并有效抑制了青枯病的发生（P<0.05）。微生物群落分析显示，处理组根际土壤微生物Simpson多样性指数和Pielou均匀度指数显著降低。宏基因组功能分析表明，处理组在碳水化合物代谢通路上的表达显著上调（P<0.05），具体表现为果糖与甘露糖代谢、半胱氨酸与甲硫氨酸代谢、以及丙氨酸、天冬氨酸和谷氨酸代谢通路的基因丰度显著增加，包括海藻酸盐合成的关键基因（manB、alg44、algG）及氨基酸代谢基因（asdA、asnA）。结论大球盖菇基质通过提高土壤有机质和氮磷含量、降低速效钾含量来优化土壤理化性质；通过定向选择作用富集有益菌群，重塑根际微生态结构；同时激活果糖、甘露糖及氨基酸等关键代谢通路，抑制病原菌生物膜形成并增强植株免疫与生长，从而显著降低烟草青枯病的发病率。

关键词: 青枯病, 蘑菇基质, 根际微生物, 宏基因组, 代谢通路

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

胡阔均, 黄晓辉, 黄谊, 张宇宇, 邓征宇, 郭军, 曾银, 尹华群, 周向平, 孟德龙. 大球盖菇基质对烟草青枯病和土壤微生物功能的影响[J]. 生物技术通报, doi: 10.13560/j.cnki.biotech.bull.1985.2025-1210.

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

图/表 7

表1 对照组与大球盖菇基质处理组土壤理化性质和病情指数

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	*

图1 对照组与大球盖菇基质处理组多样性指数A：对照组与大球盖菇基质处理组根际微生物群落Simpson多样性指数；B：对照组与大球盖菇基质处理组根际微生物群落Pielou均匀度指数；C：对照组与大球盖菇基质处理组根际微生物群落Bray-Curtis距离；D：对照组与大球盖菇基质处理组根际微生物群落主坐标分析（PCoA）图。图中不同小写字母（a、b）表示处理间差异显著（P<0.05），相同字母表示差异不显著。下同

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

图2 对照组与大球盖菇基质处理组根际土壤微生物属水平相对丰度堆积图

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

图3 对照组与大球盖菇基质处理组KEGG通路丰度汇总及通路差异分析A：对照组与大球盖菇基质处理组KEGG一级通路RPKM（reads per kilobase per million mapped reads）汇总；B：对照组与大球盖菇基质处理组KEGG二级通路RPKM汇总及差异性；C：对照组与大球盖菇基质处理组KEGG三级通路RPKM汇总及差异性

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

图4 对照组与大球盖菇基质处理组群落功能多样性A：对照组与大球盖菇基质处理组群落功能Simpson 多样性指数；B：对照组与大球盖菇基质处理组群落功能布雷-柯蒂斯距离；C：对照组与大球盖菇基质处理组群落功能主坐标分析图（PCoA）

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

图5 果糖与甘露糖代谢通路（A）、半胱氨酸和甲硫氨酸代谢通路（B）以及丙氨酸、天冬氨酸和谷氨酸代谢通路（C）

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

图6 Mantel检验与Pearson相关性热图DI：病情指数；Ralstonia：青枯雷尔氏菌；Ko00051：果糖与甘露糖代谢通路；Ko00250：半胱氨酸和甲硫氨酸代谢通路；Ko00270：丙氨酸、天冬氨酸和谷氨酸代谢通路

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

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