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. 2022 Nov 8:13:887880.
doi: 10.3389/fmicb.2022.887880. eCollection 2022.

Diversity of endophytic fungi isolated from different plant parts of Acacia mangium, and antagonistic activity against Ceratocystis fimbriata, a causal agent of Ceratocystis wilt disease of A. mangium in Malaysia

Mohd Farid Ahmad  1 , Rozihawati Zahari  2 , Mastura Mohtar  3 , Wan Azhar Wan-Muhammad-Azrul  4 , Muhammad Syahmi Hishamuddin  2 , Nik Iskandar Putra Samsudin  5   6 , Affendy Hassan  7 , Razak Terhem  2
Affiliations

Diversity of endophytic fungi isolated from different plant parts of Acacia mangium, and antagonistic activity against Ceratocystis fimbriata, a causal agent of Ceratocystis wilt disease of A. mangium in Malaysia

Mohd Farid Ahmad et al. Front Microbiol. .

Abstract

Acacia mangium is an important wood for commercial products especially pulp and medium-density fibreboard. However, it is susceptible to Ceratocystis fimbriata infection, leading to Ceratocystis wilt. Therefore, the present work aimed to (i) establish the diversity of endophytic fungi in different plant parts of A. mangium,and (ii) evaluate the antifungal potentials of the isolated and identified endophytic fungi against C. fimbriata. Endophytic fungal identification was conducted by PCR amplification and sequencing of the internal transcribed spacer 1 (ITS1) and ITS4 regions of nuclear ribosomal DNA. A total of 66 endophytic fungi were successfully isolated from different parts of A. mangium; leaf (21), stem (13), petiole (12), root (9), flower (6), and fruit (5). The endophytic fungal isolates belonged to Ascomycota (95.5%) and Zygomycota (4.5%). For Ascomycota 13 genera were identified: Trichoderma (28.6%), Nigrospora (28.6%), Pestalotiopsis (12.7%), Lasiodiplodia (9.5%), Aspergillus (6.3%), Sordariomycetes (3%), and Neopestalotiopsis, Pseudopestalotiopsis, Eutiarosporella, Curvularia, Fusarium, Penicillium, and Hypoxylon each with a single isolate. For Zygomycota, only Blakeslea sp. (5%) was isolated. Against C. fimbriata, Trichoderma koningiopsis (AC 1S) from stem, Nigrospora oryzae (AC 7L) from leaf, Nigrospora sphaerica (AC 3F) from the flower, Lasiodiplodia sp. (AC 2 U) from fruit, Nigrospora sphaerica (AC 4P) from petiole, and Trichoderma sp. (AC 9R) from root exhibited strong inhibition for C. fimbriata between 58.33 to 69.23%. Thus, it can be concluded that certain endophytic fungi of A. mangium have the potential to be harnessed as anti-Ceratocystis agent in future biotechnological applications.

Keywords: Acacia mangium; Ceratocystis fimbriata; Ceratocystis wilt; antagonism; endophytic fungi.

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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
Maximum likelihood (ML) phylogenetic tree based on rDNA ITS sequences of endophytic fungal isolates and fungal ITS sequences from the GenBank. ML tree was constructed using the Kimura 2-parameter (K2) model and gamma distributed (+G) model. All positions containing gaps and missing data were included for analysis. Clade supports were calculated based on 1,000 bootstrap.
Figure 2
Figure 2
Dual culture plate assay between six endophytic fungal isolates against the pathogen C. fimbriata (A). (B) Trichoderma koningiopsis AC1S – stem; (C) Nigoshora spharenica AC7L – leaf; (D) Nigoshora spharenica AC3F - flower; (E) Lasiodiplodia sp. AC2U – fruit; (F) Nigoshora spharenica AC4P-petiole; (G) Trichoderma sp. AC9R-root. The plates were cultivated for 5 days at 27°C. Radial growths were measured and interaction were observed.
Figure 3
Figure 3
Inhibition percentages (I%) of endophytic fungi against the pathogen Ceratocystis fimbriata in dual culture assay. Data are mean ± standard error (SE) of triplicates.
Figure 4
Figure 4
The inhibition percentages (I%) of endophytic fungi isolated from root (A), stem (B), petiole (C), leaf (D), flower (E), and fruit (F) against the pathogen Ceratocystis fimbriata. Data are mean ± standard error (SE) of triplicates. Means followed by the same letter in each group are not significantly different at α = 0.05 according to DuncanLSD.
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