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Ribosomal ITS sequence-directed selection for endophytic Myrothecium strains antagonistic to Ralstonia solanacearum to control patchouli bacterial wilt

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Abstract

Ralstonia solanacearum (RS) wilt is a soil-borne devastating disease that causes enormous loss to patchouli in tropical and subtropical areas of Asia. The aim of this study was to develop a feasible method to isolate antagonistic fungal endophytes against RS from patchouli and to investigate their effects on biocontrol of patchouli RS wilt. Diversity of endophytic fungi in roots, stems and leaves of healthy patchouli was estimated with Illumina-based sequencing strategies. The number of fungal operational taxonomic units (OTUs) at 97% similarity level was 465, 542 and 298 in roots, stems and leaves, respectively. The most detected genus in patchouli stems and leaves was Aspergillus, while the most dominant genera in roots were Athelia (11.1%) and Myrothecium (10.1%). Due to wide applications of Myrothecium species for biocontrol, 40 isolates of Myrothecium were isolated from patchouli roots using the selection medium for Myrothecium. At the species level, the strain (SWG1) with the most potent activity against RS in vitro was further identified and used to control patchouli RS wilt in vivo. The results of pot experiments showed that fresh weight, dry weight and height of patchouli seedlings inoculated with SWG1 increased by 58.2%, 65.5% and 11.9%, respectively, compared to seedlings without inoculation. The inoculation of SWG1 reduced the disease incidence and disease index of patchouli RS wilt by 66.7% and 71.0%, respectively. The results indicated the directed isolation of antagonistic endophytic fungi based on Illumina sequencing analysis of the ribosomal fungal ITS should be valuable to screen biological agents to control plant diseases.

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Data availability

The 16S rDNA data has been deposited in NCBI GenBank under the accession numbers of MW940739.

Code availability

Not applicable.

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Acknowledgements

We thank Prof. Renduo Zhang and Lixiang Cao (Sun Yat-sen University) for their help in the English writing of this manuscript. This work was supported by grants from the Chinese National Natural Science Foundation (Grant Number 31971384).

Funding

This work was supported by grants from the Chinese National Natural Science Foundation (Grant Number 31971384).

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Contributions

Zujun Deng and Wenguang Shi conceived and designed the experiments. Wenguang Shi, Jiaqi Li, HuaXiong Yao, Jianbin Li, Meiqi Chen, Danting Chen, Yinghao Fu, Huarong Xie performed the experiments. Wenguang Shi analyzed the data. Wenguang Shi and Zujun Deng wrote the manuscript with critical input from all authors. All authors read and approved the manuscript.

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Correspondence to Zujun Deng.

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The authors declare that they have no conflict of interests. All authors edited the manuscript and approved the final version.

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Handling Editor: Sotiris Tjamos

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Shi, W., Li, J., Yao, H. et al. Ribosomal ITS sequence-directed selection for endophytic Myrothecium strains antagonistic to Ralstonia solanacearum to control patchouli bacterial wilt. BioControl 67, 237–248 (2022). https://doi.org/10.1007/s10526-021-10125-3

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