ASE|栽培灵芝绿霉病发生的根际生态学机制

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来源：生态学与环境

2025-12-12 16:14:58

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核心提示：本研究为灵芝病害的管理以及灵芝的可持续生产提供了新的见解。

作物连作障碍

论文概要

研究背景

早在宋代，《重修政和经史证类备用本草》中就有“赤芝生霍山”的明确记载，印证了安徽霍山、金寨一带作为赤芝道地产区的悠久渊源。东汉《神农本草经》将灵芝列为上品药材，强调其“益心气、补中、增聪明”的药用价值，而李时珍《本草纲目》更明确大别山区是赤芝的重要发源地。1988年，金寨县沙河乡率先建起灵芝药材生产基地，标志着安徽开启规模化椴木栽培灵芝的序幕。1993年，短椴木灵芝引种栽培获得成功，技术突破推动产业规模快速扩张，霍山、岳西等多地随后建成标准化生产基地，其中霍山县1997年实现赤灵芝大面积种植，形成多点布局的产业格局。然而，椴木灵芝栽培过程中会遭受绿霉病（又称青霉病）的侵害，其主要由木霉菌等真菌引起，对灵芝产量与品质影响显著，造成严重经济损失。

微生物群是宿主不可分割的组成部分，在作物生长和健康中发挥着重要作用。越来越多的证据表明，植物根际招募了有益的微生物来抑制土壤病害菌的传播。虽然土传病害与微生物的相互关系在植物中已得到广泛研究，但对于真菌系统中宿主-微生物组-病原菌相互作用的了解却很少。而灵芝作为一种药用真菌，在传统中医中具有重要价值。因此，探究栽培灵芝的微生物群落及其与病原菌的相互关系，不仅有助于理解栽培灵芝的发病机制，也为精准制定和实施生物防治策略提供重要支持。

01 主要结果

本研究采用16S、ITS 和 18S 扩增子测序技术，对健康和患病灵芝根际土壤、菌丝际土壤和灵芝内生菌中的细菌、真菌和原生动物群落进行了分析，并分析了绿霉病对灵芝微生物群落组装、共现网络的影响，以及木霉属真菌侵染灵芝的潜在机制。研究结果表明，在灵芝健康和患病状态下相关的微生物群落表现出显著差异。中性群落模型结果显示，在健康的灵芝中，微生物群落的组装主要受随机过程控制，而在患病的灵芝中，确定性过程则更为突出。分子生态网络结果表明，与患病的灵芝相比，健康的灵芝共现网络表现出更高的稳定性，这可能归因于其对病原体诱导的压力具有更强的抵抗力。相反，患病灵芝的共现网络表现出更高的复杂性，并能招募有益细菌（Pseudomonas ASV2 和ASV22）和消费者原生动物（Proleptomons ASV28和Colpoda ASV240）。此外，体外实验还表明灵芝多糖和三萜类化合物能显著促进灵芝致病菌（木霉菌）的生长。本研究揭示了宿主微生物组在灵芝菌丝体深层培养中的重要作用，为宿主、细菌、真菌和原生动物之间复杂的相互作用提供了有价值的见解，为利用微生物资源提高农业生产力提供新策略。

Fig. 1. Differences in microbial community composition between the healthy and diseased G. lucidum at the genus level. A, B and C, Relative abundances of bacterial (A), fungal (B) and protistan (C) genus shown in Stacked graphs. The relative abundance of bacterial (D, G and J), fungal (E, H and K), and protistan (F, I and L) genus on the three sampling sites (bulk soils, hyphosphere soils, and endosphere of G. lucidum, respectively) using Statistical Analysis of Metagenomic Profiles (STAMP). P < 0.05 indicates significant differences between the healthy and diseased G. lucidum. Error bars represent Welch's t-interval. * means P < 0.05; ** means P < 0.01, *** means P < 0.001. HB and DB: Bulk soil of healthy and diseased G. lucidum; HH and DH: Hyphosphere soil of healthy and diseased G. lucidum; HE and DE: Endosphere of healthy and diseased G. lucidum.

Fig. 2. Manhattan plot showing the differential abundance ASVs enriched or depleted in the bacterial (A), fungal (B), and protistan (C) communities between the healthy and diseased G. lucidum on the three niches (bulk soil, hyphosphere soil, and endosphere of G. lucidum, respectively). Each dot or triangle represents each ASV (ASV abundance >0.01 %, P < 0.05, FDR adjustment). ASVs were colored according to the taxonomic family. The size of each dot or triangle represents the relative abundance of each ASV. Solid upward triangles indicate the ASVs enriched in the diseased G. lucidum. Hollow downward triangles represent the ASVs depleted in the diseased G. lucidum.

Fig. 3. Fit of the NCMs of the bacterial (A, B), fungal (C, D), and protistan (E, F) communities assemblage between the healthy and diseased G. lucidum. The solid blue lines indicate the best fit to the NCM, and the dashed blue lines represent 95 % confidence intervals around the model prediction. ASVs that occur more or less frequently than those predicted by the NCM are shown in different colors. R2 indicates the fit of the model. Nm denoted the metacommunity size times immigration. m indicated estimated migration rate.

Fig. 5. Intra-kingdom co-occurrence networks in the bulk soils, hyphosphere soils, and hyphal endosphere. A, The co-occurrence networks of the microbial communities between the healthy and diseased G. lucidum. The nodes are colored according to taxonomic information at the phylum level. Node size indicates the degree of connection. Edge colour represents positive (green) and negative (red) correlations. B and C, The microbial communities node-level co-occurrence networks topological properties (degree and clustering coeffcient) of the healthy and diseased G. lucidum. ns indicates no significant difference; * means P < 0.05; ** means P < 0.01, *** means P < 0.001. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Fig. 6. Interkingdom co-occurrence networks. A, Interdomain ecological networks of the bacterial-fungal-protistan associations of the healthy and diseased G. lucidum. The nodes were colored according to the different kingdom (bacteria in yellow, fungi in purple and protists in blue). Green numbers inside parentheses represent positive edge numbers and red numbers inside parentheses represent negative edge numbers. The networks in the bulk soil, hyphosphere soil, and endosphere of G. lucidum are presented in Fig. S5. B, Number of nodes for bacterial, fungal and protistan taxa in the healthy and diseased networks. C, Number of correlations in the healthy and diseased networks. The green and red lines signify positive and negative correlations, respectively. D and E, Degree (D) and clustering coefficient (E) values in the healthy and diseased networks. F, Contributions of keystone taxa to the dissimilarities of microbial communities based on correlation and best multiple regression model. *** means P < 0.001. Circle size represents the variable importance (that is, proportion of explained variability calculated via multiple regression modeling and variance decomposition analysis). Colors represent Spearman correlations.

02 主要结论

椴木栽培灵芝在健康和患病状态下相关的微生物群落表现出显著差异；我们的研究还发现，在不同的采样点，细菌、真菌和原生动物受到灵芝病害的影响程度不同，灵芝内生微生物可能首先对灵芝病害做出反应。此外，通过提取物添加实验，我们还研究了木霉菌侵染灵芝的可能机制，发现灵芝提取物在体外可被病原体利用，并与微生物群落的变化有关。总之，本研究为灵芝病害的管理以及灵芝的可持续生产提供了新的见解。