Skip to main content
SpringerLink
Log in

Isolation and partial characterization of Bacillus subtilis ME488 for suppression of soilborne pathogens of cucumber and pepper

  • Applied Microbial and Cell Physiology
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

Environmentally friendly control measures are needed for suppression of soilborne pathogens of vegetable crops in the Republic of Korea. In vitro challenge assays were used to screen approximately 500 bacterial isolates from 20 Korean greenhouse soils for inhibition of diverse plant pathogens. One isolate, Bacillus subtilis ME488, suppressed the growth of 39 of 42 plant pathogens tested. Isolate ME488 also suppressed the disease caused by Fusarium oxysporum f. sp. cucumerinum on cucumber and Phytophthora capsici on pepper in pot assays. Polymerase chain reaction was used to screen isolate ME488 for genes involved in biosynthesis of 11 antibiotics produced by various isolates of B. subtilis. Amplicons of the expected sizes were detected for bacD and bacAB, ituC and ituD, and mrsA and mrsM involved in the biosynthesis of bacilysin, iturin, and mersacidin, respectively. The identity of these genes was confirmed by DNA sequence analysis of the amplicons. Bacilysin and iturin were detected in culture filtrates from isolate ME488 by gas chromatography coupled with mass spectroscopy and by thin layer chromatography, respectively. Detection of mersacidin in ME488 culture filtrates was not attempted. Experiments reported here indicate that B. subtilis ME488 has potential for biological control of pathogens of cucumber and pepper possibly due to the production of antibiotics.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1

Similar content being viewed by others

References

  • Ahn IP, Chung HS, Lee YH (1998) Vegetative compatibility groups and pathogenicity among isolates of Fusarium oxysporum f. sp. cucumerinum. Plant Dis 82:244–246

    Article  Google Scholar 

  • Altena K, Guder A, Cramer C, Bierbaum G (2000) Biosynthesis of the lantibiotic mersacidin: organization of a Type B lantibiotic gene cluster. Appl Environ Microbiol 66:2565–2571

    Article  Google Scholar 

  • Armstrong GM Armstrong GK (1978) Formae speciales and races of Fusarium oxysporum causing wilts of the Cucurbitacea. Phytopathology 68:19–28

    Google Scholar 

  • Brannen PM Kenney DS (1997) Kodiak-A successful biological-control product for suppression of soil-borne plant pathogens of cotton. J Ind Microbiol Biotech 19:169–171

    Google Scholar 

  • Chae DH, De Jin R, Hwangbo H, Kim YW, Kim YC, Park RD, Krishnan HB, Kim KY (2006) Control of late blight (Phytophthora capsici) in pepper plants with a compost containing a multitude of chitinase-producing bacteria. BioControl 51:339–351

    Article  Google Scholar 

  • Chellemi DO (2006) Effect of urban plant debris and soil management practices on plant parasitic nematodes, Phytophthora blight and Pythium root rot of bell pepper. Crop Prot 25:1109–1116

    Article  Google Scholar 

  • Cho S-J, Lee SK, Byeong JC, Kim YH, Shin K-S (2003) Detection and characterization of the Gleosporium gleosporioides growth inhibitory compound iturin A from Bacillus subtilis strain KS03. FEMS Microbiol Lett 223:47–51

    Article  Google Scholar 

  • Chung S, Kim S-D (2005) Biological control of phytopathogenic fungi by Bacillus amyloliquefaciens 7079 provides suppression rates superior to chemical fungicides. J Microbiol Biotechnol 15:1011–1021

    Google Scholar 

  • Chung S, Kim S-D (2007) Escherichia coli can produce recombinant chitinase in the soil to control pathogenesis by Fusarium oxysporum without colonization. J Microbiol Biotechnol 17:474–480

    Google Scholar 

  • Govan J, Deretic V (1996) Microbial pathogenesis in cystic fibrosis: mucoid Pseudomonas aeruginosa and Burkholderia cepacia. Microbiol Rev 60:539–574

    PubMed Central  Google Scholar 

  • Hsieh F-C, Li M-C, Lin T-C, Kao S-S (2004) Rapid detection and characterization of surfactin-producing Bacillus subtilis and closely related species based on PCR. Curr Microbiol 49:186–191

    Article  Google Scholar 

  • Jacobsen BJ, Zidack NK, Larson BJ (2004) The role of Bacillus-based biological control agents in integrated pest management systems: plant diseases. Phytopathology 94:1272–1275

    Article  Google Scholar 

  • Kim KD, Nemec S, Musson G (1997) Effects of composts and soil amendments on soil microflora and Phytophthora root and crown rot of bell pepper. Crop Prot 16:165–172

    Article  Google Scholar 

  • Lamour KH, Hausbeck MK (2001a) The dynamics of Mefenoxam insensitivity in a recombining population of Phytophthora capsici characterized with amplified fragment length polymorphism markers. Phytopathology 91:553–557

    Google Scholar 

  • Lamour KH, Hausbeck MK (2001b) Investigating the spatiotemporal genetic structure of Phytophthora capsici in Michigan. Phytopathology 91:973–980

    Google Scholar 

  • Larkin RP, Roberts DP, Gracia-Garza JA (1998) Biological control of fungal diseases. In: Hutson D, Miyamoto J (eds) Fungicidal activity. Chemical and biological approaches to plant protection. Wiley, New York, p 149–191

    Google Scholar 

  • Mao W, Lewis JA, Lumsden RD, Hebbar KP (1998) Biocontrol of selected soilborne diseases of tomato and pepper plants. Crop Prot 17:535–542

    Article  Google Scholar 

  • Martin FN (2003) Development of alternative strategies for management of soilborne pathogens currently controlled with methyl bromide. Annu Rev Phytopathol 41:325–350

    Article  Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Mitchell DJ, Kannwischer-Mitchell ME (1992) Phytophthora. In: Singleton LL, Mihail JD, Rush CM (eds) Methods for research on soilborne phytopathogenic fungi. American Phytopathological Society, St Paul, MN, p 31–38

    Google Scholar 

  • Myung I-S, Hong S-G, Lee Y-G, Choi H-W, Shim H-S, Park J-W, Park K-S, Lee S-Y, See S-D, Lee S-H, Choi H-S, Kim Y-G, Shin D-B, Yae W-H, Han S-S, Cho W-D (2006) Review of disease incidences of major crops of the South Korea in 2005. Res Plant Dis 12:153–157

    Article  Google Scholar 

  • Parra G, Ristaino JB (2001) Resistance to mefenoxam and metalaxyl among field isolates of Phytophthora blight of bell pepper. Plant Dis 85:1069–1075

    Article  Google Scholar 

  • Pavlou GC, Vakalounakis DJ (2005) Biological control of root and stem rot of greenhouse cucumber caused by Fusarium oxysporum f. sp. radicis-cucumerinum, by lettuce soil amendment. Crop Prot 24:135–140

    Article  Google Scholar 

  • Pavlou GC, Vakalounakis DJ, Ligoxigakis EK (2002) Control of root and stem rot of cucumber, caused by Fusarium oxysporum f. sp. radicis-cucumerinum, by grafting onto resistant rootstocks. Plant Dis 86:379–382

    Article  Google Scholar 

  • Rajkovic A, Uyttendaele M, Ombregt SA, Jaaskelainen E, Salkinoja-Salonen M, Debevere J (2006) Influence of type of food on the kinetics and overall production of Bacillus cereus emetic toxin. J Food Prot 69:847–852

    Article  Google Scholar 

  • Ristaino JB, Johnston SA (1999) Ecologically based approaches to management of Phytophthora blight on bell pepper. Plant Dis 83:1080–1089

    Article  Google Scholar 

  • Roscoe J, Abraham EP (1966) Experiments relating to biosynthesis of bacilysin. Biochem J 99:793–800

    PubMed  PubMed Central  Google Scholar 

  • Rozen S, Skaletsky HJ (2000) Primer3 on the WWW for general users and for biologist programmers. In: Krawetz S, Misener S (eds) Bioinformatics methods and protocols: methods in molecular biology. Humana, Totowa, New Jersey, p 365–386

    Google Scholar 

  • Russell DW (1960) Révélation des N-méthylaminoacides. J Chromatogr 4:251–252

    Article  Google Scholar 

  • Sambrook J, Russell DW (2001) Molecular cloning. A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor New York

    Google Scholar 

  • Sherf AF, MacNab AA (1986) Fusarium wilt of muskmelon. In: Vegetable diseases and their control, 2nd ed. Wiley, New York, pp 334–337

  • Stein T (2005) Bacillus subtilis antibiotics: structures, syntheses and specific functions. Mol Microbiol 56:845–857

    Google Scholar 

  • Vakalounakis DJ (1996) Allelism of the Fcu-1 and Foc genes conferring resistance to Fusarium wilt in cucumber. Eur J Plant Pathol 102:855–858

    Article  Google Scholar 

  • Walker JE, Abraham EP (1970) The structure of bacilysin and other products of Bacillus subtilis. Biochem J 118:563–570

    PubMed Central  Google Scholar 

  • Weisburg WG, Barns SM, Lelletier DA, Lane DJ (1991) 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 173:697–670

    Article  Google Scholar 

Download references

Acknowledgements

The authors are grateful to K.-M. Kim, Yeungnam University; K.-C. Park, Gyeongbuk Agricultural Research and Extension Services; and Laurie McKenna, Sustainable Agricultural Systems Laboratory, USDA–ARS. This research was supported in part by the Korean Innovative Technology Fund from the Small and Mid-Size Business Administration to S. Chung (S0050382). A Korean patent was obtained for B. subtilis ME488 (Patent Registration Number: 10-045725-00-00).

Author information

Authors and Affiliations

  1. Department of Applied Microbiology, Yeungnam University, Gyeongsan, 712-749, Gyeongbuk, Republic of Korea

    Soohee Chung & Sang-Dal Kim

  2. Laboratory of Methods Development, CBER, FDA, Rockville, MD, 20852, USA

    Hyesuk Kong

  3. Sustainable Agricultural Systems Laboratory, USDA—Agricultural Research Service, Bldg. 001, Room 140, Beltsville Agricultural Research Service, 10300 Baltimore Avenue, Beltsville, MD, 20705-2350, USA

    Jeffrey S. Buyer, John Lydon & Daniel P. Roberts

  4. Florist and Nursery Plants Unit, USDA—Agricultural Research Service, Beltsville, MD, 20705, USA

    Dilip K. Lakshman

Authors
  1. Soohee Chung
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hyesuk Kong
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jeffrey S. Buyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Dilip K. Lakshman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. John Lydon
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Sang-Dal Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Daniel P. Roberts
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Daniel P. Roberts.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Chung, S., Kong, H., Buyer, J.S. et al. Isolation and partial characterization of Bacillus subtilis ME488 for suppression of soilborne pathogens of cucumber and pepper. Appl Microbiol Biotechnol 80, 115–123 (2008). https://doi.org/10.1007/s00253-008-1520-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-008-1520-4

Keywords

Search

Navigation