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. 2023 Jan 28;9(2):175.
doi: 10.3390/jof9020175.

The Effect of Trichoderma harzianum Hypovirus 1 (ThHV1) and Its Defective RNA ThHV1-S on the Antifungal Activity and Metabolome of Trichoderma koningiopsis T-51

Jiaqi You  1 , Zheng Hu  2 , Chaohan Li  1 , Hongjuan Yang  1 , Lihua Zhu  1 , Biting Cao  1 , Ronghao Song  3 , Weihong Gu  1
Affiliations

The Effect of Trichoderma harzianum Hypovirus 1 (ThHV1) and Its Defective RNA ThHV1-S on the Antifungal Activity and Metabolome of Trichoderma koningiopsis T-51

Jiaqi You et al. J Fungi (Basel). .

Abstract

Mycoviruses widely exist in filamentous fungi and sometimes cause phenotypic changes in hosts. Trichoderma harzianum hypovirus 1 (ThHV1) and its defective RNA ThHV1-S were found in T. harzianum and exhibited high transmissibility. In our previous study, ThHV1 and ThHV1-S were transferred to an excellent biological control agent T. koningiopsis T-51 to form a derivative strain 51-13. In this study, we assessed the metabolic changes in strain 51-13 and antifungal activity of its culture filtrate (CF) and volatile organic compounds (VOCs). The antifungal activity of CF and VOCs of T-51 and 51-13 was different. Compared with the CF of T-51, that of 51-13 exhibited high inhibitory activity against B. cinerea, Sclerotinia sclerotiorum, and Stagonosporopsis cucurbitacearum but low inhibitory activity against Leptosphaeria biglobosa and Villosiclava virens. The VOCs of 51-13 exhibited high inhibitory activity against F. oxysporum but low inhibitory activity against B. cinerea. The transcriptomes of T-51 and 51-13 were compared; 5531 differentially expressed genes (DEGs) were identified in 51-13 with 2904 up- and 2627 downregulated genes. In KEGG enrichment analysis, 1127 DEGs related to metabolic pathways (57.53%) and 396 DEGs related to biosynthesis of secondary metabolites (20.21%) were clearly enriched. From the CF of T-51 and 51-13, 134 differential secondary metabolites (DSMs) were detected between T-51 and 51-13 with 39 up- and 95 downregulated metabolites. From these, 13 upregulated metabolites were selected to test their antifungal activity against B. cinerea. Among them, indole-3-lactic acid and p-coumaric acid methyl ester (MeCA) exhibited strong antifungal activity. The IC50 of MeCA was 657.35 μM and four genes possibly related to the synthesis of MeCA exhibited higher expression in 51-13 than in T-51. This study revealed the mechanism underlying the increase in antifungal activity of T-51 because of the mycovirus and provided novel insights in fungal engineering to obtain bioactive metabolites via mycoviruses.

Keywords: Trichoderma; antifungal activity; biological control; metabolome; mycovirus.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Mycelial morphology of T-51 and 51-13 observed using SEM.
Figure 2
Figure 2
Inhibition of mycelial growth of various plant pathogenic fungi by the CF of T-51 and 51-13. * p < 0.05; student’s t-test.
Figure 3
Figure 3
Images obtained by scanning electron microscopy showing the mycelial morphology of B. cinerea growing on PDA media (CK) and PDA added with the CF of T-51 or 51-13 (10% v/v).
Figure 4
Figure 4
The effect of the volatile organic compounds produced by T-51 and 51-13 on the mycelial growth of B. cinerea (A) and F. oxysporum (B). Their mycelial morphology (C) was scanned under an SEM. * p < 0.05; student’s t-test. The red arrow showed the extracellular substance.
Figure 5
Figure 5
Transcriptome analysis of T-51 and 51-13. (A) Volcano plot of differentially expressed genes (DEGs); red and green dots represent significantly up- and downregulated genes, respectively. (B) Heatmap of the DEGs based on hierarchical clustering analysis. (C) Top 50 terms of the gene ontology (GO) enrichment of the DEGs. The percentage of the column represents the ratio of the enriched DEGs to the total annotated genes. (D) KEGG enrichment of the DEGs. Rich factor represents the ratio of the enriched DEGs to the total genes annotated in the corresponding pathway. (E) KEGG classification of the DEGs. The percentage of the column represents the ratio of the enriched DEGs to the total annotated genes.
Figure 6
Figure 6
Metabolomic profiling of T-51 and 51-13. (A) Volcano plot of differential secondary metabolites (DSMs). Red and green dots represent the significantly up- and downregulated metabolites, respectively. (B) Heatmap of the DSMs based on hierarchical clustering analysis. (C) KEGG enrichment of the DSMs. Rich factor represents the ratio of the enriched DSMs to the total metabolites annotated in the corresponding pathway.
Figure 7
Figure 7
Effect of 13 compounds (A–M; Table 1; 1 mM) on the mycelial growth of B. cinerea. The same lowercase letters indicate that the difference was not significant at p < 0.05, according to Duncan’s multiple range tests.
Figure 8
Figure 8
Inhibition curve of p-coumaric acid methyl ester against B. cinerea.
Figure 9
Figure 9
Relative quantification of four genes probably involved in the synthesis of p-coumaric acid methyl ester, which were selected by upregulation in transcriptome analysis. The data are expressed as means ± SD (n = 4). * p < 0.05; student’s t-test. (A) E1, (B) E2, (C) E3, (D) E4.
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