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Molecular Detection of Biosynthetic Genes for Anti-fungal Metabolite Production by Yam (Dioscorea sp.) Rhizobacteria

Received: Oct. 16, 2023    Accepted: Nov. 06, 2023    Published: Nov. 17, 2023
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Abstract

Biosynthetic genes are responsible for antibiotic production by rhizobacterial antagonists. Detection of antibiotic biosynthetic genes of an antagonist is therefore important for the identification of genes and antibiotics responsible for disease suppression. The study was carried out to detect the antibiotic producing genes of eight rhizobacteria responsible for fungal disease suppression. Eight yam (Dioscorea sp.) rhizobacterial isolates which were found to possess antifungal properties against several plant pathogenic fungi and identified as Bacillus species were tested for the presence of genes for biosynthesis of antifungal lipopeptides; bacillomycin D, iturin A, surfactin, fengycin, and aminopolyols, zwettemycin A as possible antibiotic tools for biocontrol using specific primers. The detection of bacillomycin D gene by PCR amplification, gene sequencing, and BLAST analysis, was achieved through the use of the primer pair, BACC1-F/BACC1-R, capable of detecting 875-bp region, iturin A through the use of ITUD1-F/ITUD1-R primer pair, capable of detecting 647-bp region while primer pair SUR3-F/SUR3-R, capable of detecting 441-bp region was also used for the detection of surfactin. Three separate primer pairs were used for fengycin viz. FEND1-F/FEND1-R, FENA1-F/FENA1-R, and FENB2-F/FENB2-R, capable of detecting 964-bp region corresponding to fengycin D, fengycin A and fengycin B respectively. Zwettermycin A was detected through the use of ZWET-F2/ZWET-R1 primer pair, capable of detecting 1-kb region. The outcome of the study shows that all the eight rhizobacteria possessed biosynthetic genes for the production of bacillomycin D, iturin A, and surfactin, however, neither the three types of fengycin nor the zwettermycin A were detected. Sequenced data of these antibiotics have been deposited with GenBank and the following accession numbers assigned to bacillomycin D (MW263002-MW263009), iturin A (MW263010-MW263017), and surfactin (MW263018-MW263025). All the eight rhizobacteria tested were found to possess three out of the five biosynthetic genes namely bacillomycin D, iturin A and surfactin. The detection of these biosynthetic genes confirms and justify why these rhizobacteria are potential biocontrol agents of plant pathogens.

DOI 10.11648/j.plant.20231104.12
Published in Plant ( Volume 11, Issue 4, December 2023 )
Page(s) 122-129
Creative Commons

This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2024. Published by Science Publishing Group

Keywords

Antifungal, Lipidopeptides, Iturin A, Bacillomycin D, Surfactin, Fengycin and Zwettermycin A

References
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[37] Zeriouh, H., de Vicente, A., Perez-Garcia, A., Romero, D. Surfactin triggers biofilm formation of Bacillus subtilis in melon phylloplane and contributes to the biocontrol activity. Environmental Microbiology, 2014, 16: 2196–2211.
[38] Ramarathnam, R. Phyllosphere bacterial biological control of Leptosphaeria maculans, the blackleg pathogen of canola (Brassica napus L.): screening for potential antibiotic producers, investigation of the mechanism of control, biochemical detection of the antifungal compounds, and establishment of the role of antibiosis. Ph.D. thesis, University of Manitoba, Winnipeg, Manitoba, 2007.
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    Stephen, L., Richard, A. T., Dorcas, Q. M., Frederick, K., Abdulai, M., et al. (2023). Molecular Detection of Biosynthetic Genes for Anti-fungal Metabolite Production by Yam (Dioscorea sp.) Rhizobacteria. Plant, 11(4), 122-129. https://doi.org/10.11648/j.plant.20231104.12

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    ACS Style

    Stephen, L.; Richard, A. T.; Dorcas, Q. M.; Frederick, K.; Abdulai, M., et al. Molecular Detection of Biosynthetic Genes for Anti-fungal Metabolite Production by Yam (Dioscorea sp.) Rhizobacteria. Plant. 2023, 11(4), 122-129. doi: 10.11648/j.plant.20231104.12

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    AMA Style

    Stephen L, Richard AT, Dorcas QM, Frederick K, Abdulai M, et al. Molecular Detection of Biosynthetic Genes for Anti-fungal Metabolite Production by Yam (Dioscorea sp.) Rhizobacteria. Plant. 2023;11(4):122-129. doi: 10.11648/j.plant.20231104.12

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  • @article{10.11648/j.plant.20231104.12,
      author = {Larbi-Koranteng Stephen and Awuah Tuyee Richard and Quain Marian Dorcas and Kankam Frederick and Muntala Abdulai and Yükselbaba Utku},
      title = {Molecular Detection of Biosynthetic Genes for Anti-fungal Metabolite Production by Yam (Dioscorea sp.) Rhizobacteria},
      journal = {Plant},
      volume = {11},
      number = {4},
      pages = {122-129},
      doi = {10.11648/j.plant.20231104.12},
      url = {https://doi.org/10.11648/j.plant.20231104.12},
      eprint = {https://download.sciencepg.com/pdf/10.11648.j.plant.20231104.12},
      abstract = {Biosynthetic genes are responsible for antibiotic production by rhizobacterial antagonists. Detection of antibiotic biosynthetic genes of an antagonist is therefore important for the identification of genes and antibiotics responsible for disease suppression. The study was carried out to detect the antibiotic producing genes of eight rhizobacteria responsible for fungal disease suppression. Eight yam (Dioscorea sp.) rhizobacterial isolates which were found to possess antifungal properties against several plant pathogenic fungi and identified as Bacillus species were tested for the presence of genes for biosynthesis of antifungal lipopeptides; bacillomycin D, iturin A, surfactin, fengycin, and aminopolyols, zwettemycin A as possible antibiotic tools for biocontrol using specific primers. The detection of bacillomycin D gene by PCR amplification, gene sequencing, and BLAST analysis, was achieved through the use of the primer pair, BACC1-F/BACC1-R, capable of detecting 875-bp region, iturin A through the use of ITUD1-F/ITUD1-R primer pair, capable of detecting 647-bp region while primer pair SUR3-F/SUR3-R, capable of detecting 441-bp region was also used for the detection of surfactin. Three separate primer pairs were used for fengycin viz. FEND1-F/FEND1-R, FENA1-F/FENA1-R, and FENB2-F/FENB2-R, capable of detecting 964-bp region corresponding to fengycin D, fengycin A and fengycin B respectively. Zwettermycin A was detected through the use of ZWET-F2/ZWET-R1 primer pair, capable of detecting 1-kb region. The outcome of the study shows that all the eight rhizobacteria possessed biosynthetic genes for the production of bacillomycin D, iturin A, and surfactin, however, neither the three types of fengycin nor the zwettermycin A were detected. Sequenced data of these antibiotics have been deposited with GenBank and the following accession numbers assigned to bacillomycin D (MW263002-MW263009), iturin A (MW263010-MW263017), and surfactin (MW263018-MW263025). All the eight rhizobacteria tested were found to possess three out of the five biosynthetic genes namely bacillomycin D, iturin A and surfactin. The detection of these biosynthetic genes confirms and justify why these rhizobacteria are potential biocontrol agents of plant pathogens.
    },
     year = {2023}
    }
    

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  • TY  - JOUR
    T1  - Molecular Detection of Biosynthetic Genes for Anti-fungal Metabolite Production by Yam (Dioscorea sp.) Rhizobacteria
    AU  - Larbi-Koranteng Stephen
    AU  - Awuah Tuyee Richard
    AU  - Quain Marian Dorcas
    AU  - Kankam Frederick
    AU  - Muntala Abdulai
    AU  - Yükselbaba Utku
    Y1  - 2023/11/17
    PY  - 2023
    N1  - https://doi.org/10.11648/j.plant.20231104.12
    DO  - 10.11648/j.plant.20231104.12
    T2  - Plant
    JF  - Plant
    JO  - Plant
    SP  - 122
    EP  - 129
    PB  - Science Publishing Group
    SN  - 2331-0677
    UR  - https://doi.org/10.11648/j.plant.20231104.12
    AB  - Biosynthetic genes are responsible for antibiotic production by rhizobacterial antagonists. Detection of antibiotic biosynthetic genes of an antagonist is therefore important for the identification of genes and antibiotics responsible for disease suppression. The study was carried out to detect the antibiotic producing genes of eight rhizobacteria responsible for fungal disease suppression. Eight yam (Dioscorea sp.) rhizobacterial isolates which were found to possess antifungal properties against several plant pathogenic fungi and identified as Bacillus species were tested for the presence of genes for biosynthesis of antifungal lipopeptides; bacillomycin D, iturin A, surfactin, fengycin, and aminopolyols, zwettemycin A as possible antibiotic tools for biocontrol using specific primers. The detection of bacillomycin D gene by PCR amplification, gene sequencing, and BLAST analysis, was achieved through the use of the primer pair, BACC1-F/BACC1-R, capable of detecting 875-bp region, iturin A through the use of ITUD1-F/ITUD1-R primer pair, capable of detecting 647-bp region while primer pair SUR3-F/SUR3-R, capable of detecting 441-bp region was also used for the detection of surfactin. Three separate primer pairs were used for fengycin viz. FEND1-F/FEND1-R, FENA1-F/FENA1-R, and FENB2-F/FENB2-R, capable of detecting 964-bp region corresponding to fengycin D, fengycin A and fengycin B respectively. Zwettermycin A was detected through the use of ZWET-F2/ZWET-R1 primer pair, capable of detecting 1-kb region. The outcome of the study shows that all the eight rhizobacteria possessed biosynthetic genes for the production of bacillomycin D, iturin A, and surfactin, however, neither the three types of fengycin nor the zwettermycin A were detected. Sequenced data of these antibiotics have been deposited with GenBank and the following accession numbers assigned to bacillomycin D (MW263002-MW263009), iturin A (MW263010-MW263017), and surfactin (MW263018-MW263025). All the eight rhizobacteria tested were found to possess three out of the five biosynthetic genes namely bacillomycin D, iturin A and surfactin. The detection of these biosynthetic genes confirms and justify why these rhizobacteria are potential biocontrol agents of plant pathogens.
    
    VL  - 11
    IS  - 4
    ER  - 

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Author Information
  • Department of Crop and Soil Sciences, Akenten Appiah–Menka University of Skills Training and Entrepreneurial Development-AAMUSTED), Mampong Campus, Asante Mampong, Ghana

  • Crop and Soil Sciences Department, University of Science and Technology (KNUST), Kumasi, Ghana

  • Crop Research Institute (CRI), Council for Scientific and Industrial Research (CSIR), Kumasi, Ghana

  • Department of Agronomy, Faculty of Agriculture, Food and Consumer Sciences, University for Development Studies, Tamale, Ghana

  • Department of Horticulture and Crop Production, School of Agriculture and Technology, University of Energy and Natural Resources (UENR), Dormaa Ahenkro, Ghana

  • Department of Plant Protection, Faculty of Agriculture, Akdeniz University, Antalya, Turkey

  • Section