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. 2025 Mar 18:16:1570200.
doi: 10.3389/fmicb.2025.1570200. eCollection 2025.

Transcriptome analysis unveils the functional effects of ectomycorrhizal fungal colonization on cadmium tolerance of willow saplings

Lijiao Wang #  1 , Baoshan Yang #  1   2 , Hui Wang  1   2 , Jiaxing Shi  1 , Jinhao Dong  1 , Xiaoxia Zhao  3 , Guanghua Qin  4 , Xinhua He  5 , Meiyuan Wang  1
Affiliations

Transcriptome analysis unveils the functional effects of ectomycorrhizal fungal colonization on cadmium tolerance of willow saplings

Lijiao Wang et al. Front Microbiol. .

Abstract

Introduction: Ectomycorrhizal fungus (ECMF) could enhance plant tolerance to heavy metal toxicity by altering metal accumulation and protecting plants from oxidative injury. However, the molecular mechanisms underlying ECMF-mediated detoxification of cadmium (Cd) in willow sapling are not well known. This study aimed to unveil the roles of Cenococcum geophilum (CG) and Suillus luteus (SL) in regulating Cd toxicity tolerance in willow (Salix psammophila 'Huangpi1') saplings.

Methods: This study systematically evaluated physiological and biochemical parameters in the leaf and root tissues of 18 willow saplings, while concurrently conducting transcriptomic analysis of the roots under Cd stress. The specific treatments were labeled as follows: NF (no ECMF inoculation and no Cd addition), CG (CG colonization only), SL (SL colonization only), NF+Cd (no ECMF inoculation with 100 μM Cd addition), CG+Cd (CG colonization with 100 μM Cd addition), and SL+Cd (SL colonization with 100 μM Cd addition).

Results: The results showed the growth, photosynthesis, antioxidant system and transcriptome of 2-month-old willow saplings responded differently to ECMFs colonization under Cd stress. S. luteus markedly increased the aerial parts biomass, while C. geophilum significantly enhanced the root property indices of willow saplings under Cd stress. The highest number of differentially expressed genes (DEGs) was observed in the comparison between CG+Cd (CG colonization with 100 μM Cd addition) and NF+Cd (no ECMF inoculation with 100 μM Cd addition). C. geophilum colonization activated plant hormone signal transduction and carbohydrate metabolism pathways, while S. luteus enhanced the synthesis of secondary metabolites.

Discussion: This study provides a molecular perspective on the mechanism of interaction between ECMFs and willow saplings under Cd stress and supports the application of ECMFs for phytoremediation of Cd-contaminated soil.

Keywords: antioxidant defense; differentially expressed genes; physiochemical responses; secondary metabolites; willow sapling.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Effects of no-ECMF inoculation (NF), Cenococcum geophilum (CG) or Suillus luteus (SL) colonization and Cd addition (100 μM) on leaf and root CAT (A and B), POD (C and D) and SOD (E and F) activity of willow saplings. Abbreviations: CAT, Catalase; POD, Peroxidase and SOD, Superoxide dismutase. Different letters indicate a significant difference at p < 0.05 by Duncan’s multiple range test.
Figure 2
Figure 2
Effects of no-ECMF inoculation (NF), Cenococcum geophilum (CG) or Suillus luteus (SL) colonization and Cd addition (100 μM) on leaf and root MDA (A and B), GSH (C and D) and ASA (E and F) of willow sapling. Abbreviations: MDA, Malondialdehyde; GSH, Reduced glutathione and ASA, Ascorbic acid. Different letters indicate a significant difference at p < 0.05 by Duncan’s multiple range test.
Figure 3
Figure 3
Volcano plot of different groups in (A) CG + Cd vs. NF + Cd and (B) SL + Cd vs. NF + Cd. (C) Venn diagram of DEGs between two groups. NF + Cd represent no ECMF inoculation (NF) with 100 μM Cd addition; CG + Cd, Cenococcum geophilum (CG) colonization with 100 μM Cd addition; SL + Cd, Suillus luteus (SL) colonization with 100 μM Cd addition.
Figure 4
Figure 4
DEGs of GO and KEGG enrichment analysis. The enriched DEGs in GO analysis (A) CG + Cd vs. NF + Cd and (B) SL + Cd vs. NF + Cd. The enriched DEGs in KEGG analysis in (C) CG + Cd vs. NF + Cd and (D) SL + Cd vs. NF + Cd. NF + Cd represent no ECMF inoculation (NF) with 100 μM Cd addition; CG + Cd, Cenococcum geophilum (CG) colonization with 100 μM Cd addition; SL + Cd, Suillus luteus (SL) colonization with 100 μM Cd addition.
Figure 5
Figure 5
Plant hormone signaling transduction pathway in Salix psammophila ‘Huangpi1’ under Cd stress. NF + Cd represent no ECMF inoculation (NF) with 100 μM Cd addition; CG + Cd, Cenococcum geophilum (CG) colonization with 100 μM Cd addition; SL + Cd, Suillus luteus (SL) colonization with 100 μM Cd addition.
Figure 6
Figure 6
Transcriptome analysis of different treatments in Salix psammophila ‘Huangpi1’ under Cd stress. NF + Cd represent no ECMF inoculation (NF) with 100 μM Cd addition; CG + Cd, Cenococcum geophilum (CG) colonization with 100 μM Cd addition; SL + Cd, Suillus luteus (SL) colonization with 100 μM Cd addition.
Figure 7
Figure 7
Network heatmap of growth parameters and physiological indices of Salix psammophila ‘Huangpi1’. DW refers dry weight of saplings. (S) meant the aerial part and (R) meant the underground part.
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