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Elucidation of the antagonistic effect of Bacillus species against white mold fungus Sclerotinia sclerotiorum

Afsana Hossain, Rakibul Hasan, Md. Monirul Islam, Juel Datta, Md. Mahidul Islam Masum

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Int. J. Biosci.13(4), 195-207, October 2018

DOI: http://dx.doi.org/10.12692/ijb/13.4.195-207


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Sclerotinia sclerotiorum is one of the most devastating and cosmopolitan soil-borne fungus and causes white mold diseases on several economically important crops, such as oilseed rape, soybean, cotton and numerous vegetable crops. The disease has been controlled primarily through the use of several chemicals; however, the fungicides are often ineffective and not environmentally safe. The use of Bacillus species has attracted much attention because of their biocontrol potential for combating plant fungal diseases. The aims of this study were therefore to evaluate the antifungal activity of Bacillus species and to elucidate the underlying inhibitory mechanism against S. sclerotiorum. The inhibitory effect of Bacillus species on S. sclerotiorum was screened by agar diffusion assay. In dual culture agar diffusion experiment, strong antifungal activity was observed against mycelial growth of S. sclerotiorum by four isolates of Bacillus, while Bacillus amyloliquefaciens isolate A1 showed highest antifungal activity (67%). Moreover, cell free culture filtrate of isolate A1 exhibited strong mycelial growth inhibition of fungus (77.5%). Microscopic studies revealed the morphological alteration of fungal hyphae after treated with isolate A1. The abilities of biofilm, siderophore and lip peptides production were also investigated to clarify the underlying inhibitory mechanism of the most effective strains. Results indicated that isolate A1 showed prolific production of biofilm (0.2129 0f OD570) and strong siderophore activity (1.69 cm) as compared to other Bacillus isolates. Moreover, three metabolites groups namely iturin, fengycin and surfactin were detected in the single cell of isolate A1 using MALDI-TOF-MS spectrum analysis. Taken together, these results demonstrated that B. amyloliquefaciens isolate A1 may have potential as an effective biocontrol agent for fighting against white mold fungus S. sclerotiorum.


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Elucidation of the antagonistic effect of Bacillus species against white mold fungus Sclerotinia sclerotiorum

Almoneafy AA, Kakar KU, Nawaz Z, Li B, Saand MA, Chun-lan Y, Xie GL. 2014. “Tomato plant growth promotion and antibacterial related-mechanisms of four rhizobacterial Bacillus strains against Ralstonia solanacearum.” Symbiosis 63, 59-70. http://dx.doi.org/10.1007/s13199-014-0288-9

Arrebola E, Jacobs R, Korsten L. 2010. “Iturin A is the principal inhibitor in the biocontrol activity of Bacillus amyloliquefaciens PPCB004 against postharvest fungal pathogens.” Journal of Applied Microbiology 108, 386-395.  http://dx.doi.org/10.1111/j.1365-2672.2009.04438.x

Asaka O, Shoda M. 1996. “Biocontrol of Rhizoctonia solani damping-off of tomato with Bacillus subtilis RB14.” Appllied Environmental Microbiology 62, 4081-4085.

Ashwini N, Srividya S. 2014. “Potentiality of Bacillus subtilis as biocontrol agent for management of anthracnose disease of chilli caused by Colletotrichum gloeosporioides OGC1.” 3 Biotech 4, 127-136. http://dx.doi.org/10.1007/s13205-013-0134-4

Bais HP, Fall R, Vivanco JM. 2004. “Biocontrol of Bacillus subtilis against infection of Arabidopsis roots by Pseudomonas syringae is facilitated by biofilm formation and surfactin production.” Plant Physiology 134, 307-319. http://dx.doi.org/10.1104/pp.103.028712

Ben Ayed H, Hmidet N, Bechet M, Chollet M, Chataigne G, Leclere V, Jacques P, Nasri M. 2014. “Identification and biochemical characteristics of Lip opeptides from Bacillus mojavensis A21.” Process Biochemestry 49, 1699-1707. http://dx.doi.org/10.1016/j.procbio.2014.07.001

Davey ME, O’Toole GA. 2000. “Microbial biofilms: from ecology to molecular genetics.” Microbiology and Molecular Biology Reviews 64, 847-+. http://dx.doi.org/10.1128/mmbr.64.4.847-867.2000

de Boer M, Born P, Kindt F, Keurentjes JJB, Van der Sluis I, Van Loon LC, Bakker P. 2003. “Control of Fusarium wilt of radish by combining Pseudomonas putida strains that have different disease-suppressive mechanisms.” Phytopathology 93, 626-632. http://dx.doi.org/10.1094/phyto.2003.93.5.626

Dutta S, Ghosh PP,  Kuiry SP. 2009. “Stem rot, a new disease of potato in West Bengal, India.” Australian Plant Disease Notes 4, 80-81. http://dx.doi.org/10.1071/dn09034

El-shakh ASA, Kakar KU, Wang X, Almoneafy AA, Ojaghian MR, Li B, Anjum SI, Xie GL. 2015. “Controlling bacterial leaf blight of rice and enhancing the plant growth with endophytic and rhizobacterial Bacillus strains.” Toxicologicl Environmental Chemistry 97, 766-785. http://dx.doi.org/10.1080/02772248.2015.1066176

Elshakh ASA, Anjum SI, Qiu W, Almoneafy AA, Li W, Yang Z, Cui ZQ, Li B. 2016. “Controlling and Defence-related Mechanisms of Bacillus Strains Against Bacterial Leaf Blight of Rice.” Journal of Phytopathology 164, 534-546. http://dx.doi.org/10.1111/jph.12479

Fernando WGD, Nakkeeran S, Zhang Y, Savchuk S. 2007. “Biological control of Sclerotinia sclerotiorum (Lib.) de Bary by Pseudomonas and Bacillus species on canola petals.” Crop Protection 26, 100-107. http://dx.doi.org/10.1016/j.cropro.2006.04.007

Gerlagh M, Goossen-Van de Geijn HM, Fokkema NJ, Vereijken PFG. 1999. “Long-term biosanitation by application of Coniothyrium minitans on Sclerotinia sclerotiorum infected crops.” Phytopathology 89, 141-147.  http://dx.doi.org/10.1094/phyto.1999.89.2.141

Heckel DG. 2012. “Insecticide Resistance After Silent Spring.” Science 337, 1613-1614. http://dx.doi.org/10.1126/science.1226994

Jagadeesh KS, Kulkarni JH, Krishnaraj PU. 2001. “Evaluation of the role of fluorescent siderophore in the biological control of bacterial wilt in tomato using Tn(5) mutants of fluorescent Pseudomonas sp.” Current Science 81, 882-883.

Ji SH, Paul NC, Deng JX, Kim YS, Yun BS,  Yu SH. 2013. “Biocontrol Activity of Bacillus amyloliquefaciens CNU114001 against Fungal Plant Diseases.” Mycobiology 41, 234-242. http://dx.doi.org/10.5941/myco.2013.41.4.234

Kheiri A, Jorf SAM, Malihipour A, Saremi H, Nikkhah M. 2016. “Application of chitosan and chitosan nanoparticles for the control of Fusarium head blight of wheat (Fusarium graminearum) in vitro and greenhouse.” International Journal of Biological Macromolecules 93, 1261-1272.  http://dx.doi.org/10.1016/j.ijbiomac.2016.09.072

McGrath MT. 2001. “Fungicide resistance in cucurbit powdery mildew: Experiences and challenges.” Plant Disease 85, 236-245. http://dx.doi.org/10.1094/pdis.2001.85.3.236

Mizumoto S, Hirai M, Shoda M. 2006. “Production of lipopeptide antibiotic iturin A using soybean curd residue cultivated with Bacillus subtilis in solid-state fermentation.” Applied Microbiology and Biotechnology 72, 869-875. http://dx.doi.org/10.1007/s00253-006-0389-3

Mora I, Cabrefiga J, Montesinos E. 2011. “Antimicrobial peptide genes in Bacillus strains from plant environments.” International Microbiology 14, 213-223. http://dx.doi.org/10.2436/20.1501.01.151

Nelson B. 1998. “Biology of Sclerotinia.” Proceedings of the 10th International Sclerotinia workshop, Fargo, ND, USA, North Dakota State University Department of Plant Pathology, Fargo, ND (1998), 1-5.  http://dx.doi.org/10.1080/21691401.2016.1241793

Ojaghian MR. 2009. “First report of Sclerotinia sclerotiorum on potato plants in Iran.” Australian Plant Disease Notes 4, 39-41. http://dx.doi.org/10.1071/dn09016

Ojaghian MR, Zhang JZ, Zhang F, Qiu W, Li X. N, Xie GL, Zhu SJ. 2016. “Early detection of white mold caused by Sclerotinia sclerotiorum in potato fields using real-time PCR.” Mycological Progress 15, 959-965. http://dx.doi.org/10.1007/s11557-016-1222-8

Perez-Garcia A, Romero D,  de Vicente A. 2011. “Plant protection and growth stimulation by microorganisms: biotechnological applications of Bacilli in agriculture.” Current Opinion in Biotechnology 22, 187-193. http://dx.doi.org/10.1016/j.copbio.2010.12.003

Rahman MA, Kadir J, Mahmud TMM, Rahman RA, Begum MM. 2007. “Screening of antagonistic bacteria for biocontrol activities on Colletotrichum gloeosporioides in papaya.” Asian Journal of Plant Sciences 6, 12-20.

Ramzan N, Noreen N, Shahzad S. 2014. “Inhibition of in vitro growth of soil-borne pathogen by compost-inhabiting indigenous bacteria and fungi.” Pakistan Journal Botany 46, 1093-1099.

Raton TO, Giro ZG, Diaz MS, Perez SR. 2012. “In vitro growth inhibition of Curvularia gudauskasii by Bacillus subtilis.” Annual Microbiology 62, 545-551. http://dx.doi.org/10.1007/s13213-011-0290-x

Romero D, de Vicente A, Olmos JL, Davila J. C, Perez-Garcia A. 2007. “Effect of Lip opeptides of antagonistic strains of Bacillus subtilis on the morphology and ultrastructure of the cucurbit fungal pathogen Podosphaera fusca.” Journal of Applied Microbiology 103, 969-976. http://dx.doi.org/10.1111/j.1365-2672.2007.03323.x

Romero D, de Vicente A, Rakotoaly RH, Dufour SE, Veening JW, Arrebola E, Cazorla FM, Kuipers OP. 2007. “The iturin and fengycin families of Lip opeptides are key factors in antagonism of Bacillus subtilis toward Podosphaera fusca.” Molecular Plant-Microbe Interactions 20, 430-440.  http://dx.doi.org/10.1094/mpmi-20-4-0430

Santos S, Neto IFF, Machado MD, Soares H. MVM, Soares V. 2014. “Siderophore Production by Bacillus megaterium: Effect of Growth Phase and Cultural Conditions.” Applied Biochemestry Biotechnology 172, 549-560. http://dx.doi.org/10.1007/s12010-013-0562-y

Saravanakumar D, Clavorella A, Spadaro D, Garibaldi A, Gullino ML. 2008. “Metschnikowia pulcherrima strain MACH1 outcompetes Botrytis cinerea, Alternaria alternata and Penicillium expansum in apples through iron depletion.” Postharvest Biology Tecnology 49, 121-128. http://dx.doi.org/10.1016/j.postharvbio.2007.11.006

Schwyn B, Neilands JB. 1987. “Universal chemical-assay for the detection and determination of siderophores.” Analytical Biochemistry 160, 47-56.  http://dx.doi.org/10.1016/0003-2697(87)90612-9

Sharma N, Sharma S. 2008. “Control of foliar diseases of mustard by Bacillus from reclaimed soil.” Microbiological Research 163, 408-413. http://dx.doi.org/10.1016/j.micres.2006.06.011

Sotoyama K, Akutsu K, Nakajima M. 2016. “Biological control of Fusarium wilt by Bacillus amyloliquefaciens IUMC7 isolated from mushroom compost.” Journal of General Plant Pathology 82, 105-109. http://dx.doi.org/10.1007/s10327-015-0641-8

Spadaro D, Droby S. 2016. “Development of biocontrol products for postharvest diseases of fruit: The importance of elucidating the mechanisms of action of yeast antagonists.” Trends in Food Sciencs andTechnology 47, 39-49. http://dx.doi.org/10.1016/j.tifs.2015.11.003

Tendulkar SR, Saikumari YK, Patel V, Raghotama S, Munshi TK, Balaram P, Chattoo BB. 2007. “Isolation, purification and characterization of an antifungal molecule produced by Bacillus licheniformis BC98, and its effect on phytopathogen Magnaporthe grisea.” Journal of Applied Microbiology 103, 2331-2339. http://dx.doi.org/10.1111/j.1365-2672.2007.03501.x

Timmusk S, Grantcharova N, Wagner EGH. 2005. “Paenibacillus polymyxa invades plant roots and forms biofilms.” Applied and Environmental Microbiology 71, 7292-7300. http://dx.doi.org/10.1128/aem.71.11.7292-7300.2005

Yang H, Li X, Li X, Yu H, Shen Z. 2015. “Identification of lipopeptide isoforms by MALDI-TOF-MS/MS based on the simultaneous purification of iturin, fengycin, and surfactin by RP-HPLC.” Analytcal and Bioanalytical Chemistry 407, 2529-2542. http://dx.doi.org/10.1007/s00216-015-8486-8

Yu XM, Ai CX, Xin L, Zhou GF. 2011. “The siderophore-producing bacterium, Bacillus subtilis CAS15, has a biocontrol effect on Fusarium wilt and promotes the growth of pepper.” European Journal of Soil Biology 47, 138-145. http://dx.doi.org/10.1016/j.ejsobi.2010.11.001

Zohora US, Ano T, Rahman MS. 2016. “Biocontrol of Rhizoctonia solani K1 by Iturin A Producer Bacillus subtilis RB14 Seed Treatment in Tomato Plants.” Advances in Microbiology 6, 424-431.  http://dx.doi.org/10.4236/aim.2016.66042


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