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Isolation and characterization of a mycosubtilin homologue antagonizing Verticillium dahliae produced by Bacillus subtilis strain Z15

Bacillus subtilis strain Z15 (BS-Z15) was isolated from the cotton field of Xinjiang, China, and characterized as an effective biocontrol agent antagonizing plant pathogen Verticillium dahliae 991 (VD-991). However, the chemical substance produced by BS-Z15 for resistance to VD-991 remains elusive....

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Autores principales: Lin, Rongrong, Zhang, Qi, Yin, Li, Zhang, Yiwen, Yang, Qilin, Liu, Kai, Wang, Yingdian, Han, Shengcheng, Zhao, Huixin, Zhao, Heping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9191732/
https://www.ncbi.nlm.nih.gov/pubmed/35696380
http://dx.doi.org/10.1371/journal.pone.0269861
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author Lin, Rongrong
Zhang, Qi
Yin, Li
Zhang, Yiwen
Yang, Qilin
Liu, Kai
Wang, Yingdian
Han, Shengcheng
Zhao, Huixin
Zhao, Heping
author_facet Lin, Rongrong
Zhang, Qi
Yin, Li
Zhang, Yiwen
Yang, Qilin
Liu, Kai
Wang, Yingdian
Han, Shengcheng
Zhao, Huixin
Zhao, Heping
author_sort Lin, Rongrong
collection PubMed
description Bacillus subtilis strain Z15 (BS-Z15) was isolated from the cotton field of Xinjiang, China, and characterized as an effective biocontrol agent antagonizing plant pathogen Verticillium dahliae 991 (VD-991). However, the chemical substance produced by BS-Z15 for resistance to VD-991 remains elusive. Here, a serial purification methods including HCl precipitation, organic solvent extraction, and separation by semi-preparative High-Performance Liquid Chromatography were performed to obtain a single compound about 3.5 mg/L from the fermentation broth of BS-Z15, which has an antifungal activity against VD-991. Moreover, Fourier Transform Infrared spectrum, Nuclear Magnetic Resonance Spectroscopy, and Tandem Mass Spectrometry analyses were carried out to finally confirm that the active compound from BS-Z15 is a mycosubtilin homologue with C17 fatty acid chain. Genomic sequence prediction and PCR verification further showed that the BS-Z15 genome contains the whole mycosubtilin operon comprising four ORFs: fenF, mycA, mycB, and mycC, and the expression levels of mycA-N, mycB-Y and mycC-N reached a peak at 32-h fermentation. Although mycosubtilin homologue at 1 μg/mL promoted the germination of cotton seed, that with high concentration at 10 μg/mL had no significant effect on seed germination, plant height and dry weight. Furthermore, mycosubtilin homologue sprayed at 10 μg/mL on two-week-old cotton leaves promotes the expression of pathogen-associated genes and gossypol accumulation, and greatly decreases VD-991 infection in cotton with disease index statistics. This study provides an efficient purification strategy for mycosubtilin homologue from BS-Z15, which can potentially be used as a biocontrol agent for controlling verticillium wilt in cotton.
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spelling pubmed-91917322022-06-14 Isolation and characterization of a mycosubtilin homologue antagonizing Verticillium dahliae produced by Bacillus subtilis strain Z15 Lin, Rongrong Zhang, Qi Yin, Li Zhang, Yiwen Yang, Qilin Liu, Kai Wang, Yingdian Han, Shengcheng Zhao, Huixin Zhao, Heping PLoS One Research Article Bacillus subtilis strain Z15 (BS-Z15) was isolated from the cotton field of Xinjiang, China, and characterized as an effective biocontrol agent antagonizing plant pathogen Verticillium dahliae 991 (VD-991). However, the chemical substance produced by BS-Z15 for resistance to VD-991 remains elusive. Here, a serial purification methods including HCl precipitation, organic solvent extraction, and separation by semi-preparative High-Performance Liquid Chromatography were performed to obtain a single compound about 3.5 mg/L from the fermentation broth of BS-Z15, which has an antifungal activity against VD-991. Moreover, Fourier Transform Infrared spectrum, Nuclear Magnetic Resonance Spectroscopy, and Tandem Mass Spectrometry analyses were carried out to finally confirm that the active compound from BS-Z15 is a mycosubtilin homologue with C17 fatty acid chain. Genomic sequence prediction and PCR verification further showed that the BS-Z15 genome contains the whole mycosubtilin operon comprising four ORFs: fenF, mycA, mycB, and mycC, and the expression levels of mycA-N, mycB-Y and mycC-N reached a peak at 32-h fermentation. Although mycosubtilin homologue at 1 μg/mL promoted the germination of cotton seed, that with high concentration at 10 μg/mL had no significant effect on seed germination, plant height and dry weight. Furthermore, mycosubtilin homologue sprayed at 10 μg/mL on two-week-old cotton leaves promotes the expression of pathogen-associated genes and gossypol accumulation, and greatly decreases VD-991 infection in cotton with disease index statistics. This study provides an efficient purification strategy for mycosubtilin homologue from BS-Z15, which can potentially be used as a biocontrol agent for controlling verticillium wilt in cotton. Public Library of Science 2022-06-13 /pmc/articles/PMC9191732/ /pubmed/35696380 http://dx.doi.org/10.1371/journal.pone.0269861 Text en © 2022 Lin et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Lin, Rongrong
Zhang, Qi
Yin, Li
Zhang, Yiwen
Yang, Qilin
Liu, Kai
Wang, Yingdian
Han, Shengcheng
Zhao, Huixin
Zhao, Heping
Isolation and characterization of a mycosubtilin homologue antagonizing Verticillium dahliae produced by Bacillus subtilis strain Z15
title Isolation and characterization of a mycosubtilin homologue antagonizing Verticillium dahliae produced by Bacillus subtilis strain Z15
title_full Isolation and characterization of a mycosubtilin homologue antagonizing Verticillium dahliae produced by Bacillus subtilis strain Z15
title_fullStr Isolation and characterization of a mycosubtilin homologue antagonizing Verticillium dahliae produced by Bacillus subtilis strain Z15
title_full_unstemmed Isolation and characterization of a mycosubtilin homologue antagonizing Verticillium dahliae produced by Bacillus subtilis strain Z15
title_short Isolation and characterization of a mycosubtilin homologue antagonizing Verticillium dahliae produced by Bacillus subtilis strain Z15
title_sort isolation and characterization of a mycosubtilin homologue antagonizing verticillium dahliae produced by bacillus subtilis strain z15
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9191732/
https://www.ncbi.nlm.nih.gov/pubmed/35696380
http://dx.doi.org/10.1371/journal.pone.0269861
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