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Journal of Integrative Agriculture  2019, Vol. 18 Issue (9): 2080-2092    DOI: 10.1016/S2095-3119(19)62578-5
Special Issue: 植物细菌真菌合辑Plant Bacteria/Fungus
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Fermentation, formulation and evaluation of PGPR Bacillus subtilis isolate as a bioagent for reducing occurrence of peanut soil-borne diseases
Abdel-Gayed M. Ahmad1, Abo-Zaid G. Attia2, Matar S. Mohamed2, 3, Hafez E. Elsayed4
1?Onion, Garlic and Oil Crops Research Department, Plant Pathology Research Institute, Agricultural Research Center, Giza 12619, Egypt
2 Bioprocess Development Department, Genetic Engineering and Biotechnology Research Institute, City of Scientific Research and Technological Applications, New Borg El-Arab City, Alexandria 21934, Egypt
3 Chemical Engineering Department, Faculty of Engineering, Jazan University, Jazan 45142, Saudi Arabia 
4 Plant Protection and Biomolecular Diagnosis Department, Arid Lands Cultivation Research Institute, City of Scientific Research and Technological Applications, New Borg El-Arab City, Alexandria 21934, Egypt
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Four isolates of Bacillus subtilis coded, B4, B7, B8 and B10 were examined as biocontrol agents for their abilities and antagonistic effect on the in vitro growth of certain phytopathogenic fungi of peanut, Rhizoctonia solani and Sclerotium rolfsii.  Bacillus subtilis isolate B4 (GenBank accession no. EF150884) was the highly effective one for inhibiting the fungal mycelial growth.  Batch fermentation of B. subtilis isolate B4 was carried out and the maximum biomass achieved was 4.53 g L–1 at 11 h.  Bacillus subtilis isolate B4 was formulated and evaluated as a biofungicide to reduce peanut soil-borne diseases under greenhouse and field conditions at the side of Rizolex-T (fungicide) as standard.  Treatments by formulated plant growth-promoting rhizobacteria (PGPR) B. subtilis B4 and Rizolex-T in a soil infested with R. solani, S. rolfsii and mixture of them were more effective in decreasing percentage of damping-off, root and pod rot disease incidence (%) in greenhouse and open field environment during the two seasons 2015 and 2016.  Treatments by PGPR gave highly dry weight and number of healthy pods compared to control of fungi treatment which was nearby to dry weights of healthy pods achieved by treatments by Rizolex-T in a soil infested with S. rolfsii, R. solani and mixture of them.  Formulated PGPR B. subtilis B4 gave higher increasing of yield percentage than treatment by Rizolex-T in the two evaluated seasons 2015 and 2016.  It can conclude that the produced bioforumlated agent was more efficient as fungicide when compared with the other chemical synthesized fungicides, safe for human and the environment and economy. 
Keywords:  peanut        soil-borne diseases        Bacillus subtilis        biocontrol        fermentation       formulation  
Received: 25 July 2018   Accepted:
Corresponding Authors:  Correspondence Abo-Zaid G. Attia, Tel: +203-4593422, Fax: +203-4593423, E-mail:   
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Abdel-Gayed M. Ahmad, Abo-Zaid G. Attia, Matar S. Mohamed, Hafez E. Elsayed. 2019. Fermentation, formulation and evaluation of PGPR Bacillus subtilis isolate as a bioagent for reducing occurrence of peanut soil-borne diseases. Journal of Integrative Agriculture, 18(9): 2080-2092.

Abbasdokht H, Gholami A. 2010. The effect of seed inoculation (Pseudomonas putida+Bacillus lentus) and different levels of fertilizer on yield and yield components of wheat (Tritium aestivum L.) cultivars. International Journal of Agricultural and Biosystems Engineering, 4, 678–682.
Abd-Allah E F, El Didamony G. 2007. Effect of seed treatment of Arachis hypogaea with Bacillus subtilis on nodulation in biological control of southern blight (Sclerotium rolfsii) disease. Phytoparasitica, 35, 8–12.
Abd-El-Khair H, Haggag K H, Elshahawy I E. 2016. Soil application of Bacillus pumilus and Bacillus subtilis for suppression of Macrophomina phaseolina and Rhizoctonia solani and yield enhancement in peanut. International Journal of ChemTech Research, 9, 142–152.
Ahmed A M, Hoda A, Abdel-Gayed M A. 2017. Safe approach to control Fusarium oxysporum in sesame crop. Zagazig Journal of Agricultural Research, 44, 2529–2540.
Amer A G, Utkhede R S. 2007. Development of formulation of biological agents for management of root rot of lettuce and cucumber. Canadian Journal of Microbiology, 46, 809–816.
Asaka O, Shoda M. 1996. Biocontrol of Rhizoctonia solani damping-off of tomato with Bacillus subtilis RB14. Applied and Environmental Microbiology, 62, 4081–4085.
Ashour A Z A, Afify A H. 1999. Use of Bacillus strains mixture to improve biological control of cotton seedling disease. African Journal of Mycology and Biotechnolgy, 7, 109–120.
Baker R, Paulitz T C. 1996. Theoretical basis for microbial interactions leading to biological control of soil borne plant pathogens. In: Hall R, ed., Principles and Practice of Managing Soil Borne Plant Pathogens. The American Phytopathological Society, St. Paul, MN. pp. 50–79.
Chakraborty U, Chakraborty B N, Chowdhury P R, Tongden C, Basnet M. 2006. Investigation on Plant Growth Promoting Rhizobacteria of Tea Rhizosphere. 6th ed. International Workshop on PGPR, IISR, Calicut, Kerala.
Chaur T. 1998. General mechanisms of action of microbial biocontrol agents. Plant Pathology Bulletin, 7, 155–166.
Chen X H, Koumoutsi A, Scholz R, Schneider K, Vater J, Süssmuth R, Piel J, Borriss R J. 2009. Genome analysis of Bacillus amyloliquefaciens FZB42 reveals its potential for biocontrol of plant pathogens. Biotechnology, 140, 27–37.
Collee J G, Fraser A G, Marmion B P, Simmons A. 1996. Practical Medical Microbiology. 14th ed. Churchill Livingstone, New York.
Cook R J, Baker K F. 1983. The Nature and Practice of Biological Control of Plant Pathogens. 2nd ed. American Phytopathological Society, Saint Paul, MN, USA.
Damicone J P, Melouk H A. 2009. Soilborne Diseases of Peanut. Tech-Rep. EPP-7664, Oklahoma Cooperative Extension Service.
Van Dam-Mieras M C E, Jeu W H, Vries J, Currell B R, James J W, Leach C K, Patmore R A. 1992. Techniques Used in Bioproduct Analysis. Butterworth-Heinemann, Oxford.
Gabr M R, Hussein N A, Saleh O I, Khalil M A. 1998. Susceptibility of certain varieties and genotypes and control of wilt and root rot diseases of sesame attributed to Fusarium oxysporum f. sp. sesami and Macrophomina phaseolina. Egyptian Journal of Microbiology, 33, 403–428.
Hang N T, Oh S, Kim G H, Hur J, Koh Y J. 2005. Bacillus subtilis as a biocontrol agent against Botrytis cinerea in strawberries. Plant Pathology Journal, 21, 59–63.
Hussien Z N. 2011. New approaches for controlling peanut root rot and pod rots caused by Rhizoctonia solani in Egypt and Nigeria. Ph D thesis, Institute of African Research and Studies, Cairo University, Egypt.
Hussien Z N, Mahmoud E Y, Metwaly A H, Sobhy H M. 2012. Effect of some antagonistic bacteria in reducing of peanut damping-off, root and pod rot incidence caused by Rhizoctonia solani. Journal of Plant Protection and Pathology, 3, 1173–1187.
Ibrahim M M, Mahmoud E Y, Wagida A M. 2008. The ability of some antagonistic bacteria on control of peanut root rots diseases compared to fungicides efficiency. Minufiya Journal of Agriculture Research, 33, 1107–1125.
Kinsella K, Schulthess C P, Morris T F, Stuart J D. 2009. Rapid quantification of Bacillus subtilis antibiotics in the rhizosphere. Soil Biology and Biochemistry, 41, 374–379.
Kloepper J W, Ryu C M, Zhang S. 2004. Induced systemic resistance and promotion of plant growth by Bacillus spp. Phytopathology, 94, 1259–1266.
Leon M, Yaryura P M, Montecchia M S, Hernandez A I, Correa O S, Pucheu N L, Kerber N L, Garc?a A F. 2009. Antifungal activity of selected indigenous Pseudomonas and Bacillus from the soybean rhizosphere. International Journal of Microbiology, 2009, 1–9.
Mahmoud E Y. 2014. Performance of some antagonistic bacteria in minimizing occurrence of peanut damping-off, root and pod rot diseases. Egyptian journal of Phytopathology, 42, 205–220.
Mahmoud E Y, Shokry Y M, Zeinab N H. 2006. Efficiency of some antagonistic bacteria to reduce incidence of damping-off, wilt and peanut root rot. Journal of Agriculture Science Mansoura University, 31, 3525–3536.
Matar S M, El-Kazzaz S A, Wagih E E, El-Diwany A I, Hafez E E, Moustafa H E, Abo-Zaid G A, Serour E A. 2009a. Molecular characterization and batch fermentation of Bacillus subtilis as biocontrol agent, II. Biotechnology, 8, 35–43.
Matar S M, El-Kazzaz S A, Wagih E E, El-Diwany A I, Moustafa H E, Abo-Zaid G A, Abd-Elsalam H E, Hafez E E. 2009b. Antagonistic and inhibitory effect of Bacillus subtilis against certain plant pathogenic fungi. Biotechnology, 8, 53–61.
Maurhofer M, Keel C, Hass D, Defago G. 1995. Influence of plant species on disease suppression by Pseudomonas fluorescens strain CHAO with enhanced antibiotic production. Plant Pathology, 44, 40–50.
Meena B, Marimuthu T, Vidhyasekaran P, Velazhahan R. 2001. Biological control of root rots of groundnut with antagonistic Pseudomonas fluorescens strains. Zeitschrift für Pflanzenkrankheiten und Pflanzenschutz, 108, 369–381.
Mishra D S, Kumar A, Prajapati S E, Singh A K, Sharma S D. 2013. Identification of the compatible bacterial and fungal isolates and their effectiveness against plant diseases. Journal of Environmental Biology, 34, 183–189.
Nagorska K, Bikowski M, Obuchowski M. 2007. Multicellular behavior and production of a wide variety of toxic substances support usage of Bacillus subtilis as a powerful biocontrol agent. Acta Biochimica Polonica, 54, 495–508.
Nandakumar R, Babu S, Viswanathan R, Sheela J, Raguchander T, Samiyappan R. 2001. A new bio-formulation containing plant growth promoting rhizobacterial mixture for the management of sheath blight and enhanced grain yield in rice. Biocontrol, 46, 493–510.
Nemec S, Datnoff L E, Strandberg J. 1996. Efficacy of biocontrol agents in planting mixes to colonize plant roots and control root diseases of vegetables and citrus. Crop Protection, 15, 735–742.
Ongena M, Jourdan E, Adam A, Paquot M, Brans A, Joris B, Arpigny J L, Thornart T. 2007. Surfactin and fengycin lipopeptides of Bacillus subtilis as elicitors of induced systemic resistance in plants. Environmental Microbiology, 9, 1084–1090.
Peypoux F, Bonmatin J M, Wallach J. 1999. Recent trends in the biochemistry of surfactin. Applied Microbiology and Biotechnology, 51, 553–563.
Romero D, De Vicente A, Rakotoaly R H, Dufour S E, Veening J W, Arrebola E, Cazorla F M, Kuipers O P, Paquot M, Pérez-García A. 2007. The iturin and fengycin families of lipopeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca. Molecular Plant-Microbe Interactions, 20, 430–440.
Sailaja P R, Podile A R. 1998. A phytoalexin is modified to less fungitoxic substances by crown rot pathogen in groundnut (Arachis hypogaea L.). Indian Journal of Experimental Biology, 36, 631–634.
Sailaja P R, Podile A R, Reddauna P. 1998. Biocontrol strain of Bacillus subtilis AF1 rapidly induces lipoxygenase in groundnut Arachis hypogea L. compared with crown rot pathogen Aspergillus niger. Europian Journal of Plant Pathology, 104, 125–132.
Siddiqui Z. 2006. PGPR: Biocontrol and Biofertilization. Springer, The Netherlands.
Sneath P H A, Mair N S, Sharpe M E, Holt J G. 1986. Bergey’s manual of systematic bacteriology. In: Endospore-Forming Gram-Positive Rods and Cocci. Williams & Wilkins, Baltimore, USA. pp. 1104–1139.
Stein T. 2005. Bacillus subtilis antibiotics: Structures, syntheses and specific functions. Molecular Microbiology, 56, 845–857.
Tsuge K, Akiyama T, Shoda M J. 2001. Cloning, sequencing, and characterization of the iturin a operon. Journal of Bacteriology, 183, 6265–6273.
Vidhyasekaran P, Muthamilan M. 1995. Development of formulation of Pseudomonas fluorescens for control of chickpea wilt. Plant Disease, 79, 782–786.
Wei Y H, Wang L F, Chang J S, Kung S S. 2003. Identification of induced acidification in iron-enriched cultures of Bacillus subtilis during biosurfactant fermentation. Journal of Bioscience and Bioengineering, 96, 174–178.
Yeh M, Wei Y, Chang J. 2006. Bioreactor design for enhanced carrier-assisted surfactin production with Bacillus subtilis. Process Biochemistry, 4, 1799–1805.
Yi G, Liu Q, Lin J, Wang W, Huang H, Lia S. 2016. Repeated batch fermentation for surfactin production with immobilized Bacillus subtilis BS–37: Two-stage pH control and foam fractionation. Journal of Chemicals Technology and Biotechnology, 92, 530–535.
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