Abstract
Fire blight caused by Erwinia amylovora is an important disease affecting most types of rosaceous trees and represents a significant threat to fruit cultivation in many parts of the world. The effectiveness of antagonistic bacteria Bacillus amyloliquefaciens strain MB40 to inhibit the growth of Erwinia amylovora strain 1E IMIV was evaluated under in vitro conditions. B. amyloliquefaciens MB40 was isolated from leaves of fire blight host mature apple trees (cultivar ‘Zarya Alatau’) sampled in the Almaty region of Kazakhstan. Local isolate of the fire blight causative agent E. amylovora was used as pathogen inoculum in laboratory trials. Erwinia amylovora and Bacillus amyloliquefaciens were identified using morphological and biochemical tests, and the pathogenicity of E. amylovora 1E IMIV was demonstrated in a bioassay on immature pear fruits. B. amyloliquefaciens MB40 showed high antagonistic activity against the pathogen. Biological efficiency of the applied culture broth of B. amyloliquefaciens was 90.6% (95% confidence interval [CI] = 74.1 to 107.2). Analysis of active substances produced by the antagonistic bacterium disclosed high amounts of acetoin (3-hydroxybutanone) - 53.7 ± 1.8% and and 2,3-butanedione - 34.4 ± 3.6%. The inhibitory activity of the latter was demonstrated. The possible mechanism of its action is discussed, suggesting that B. amyloliquefaciens MB40 is a novel antibacterial strain with potential application in controlling E. amylovora in fruit growing.
Similar content being viewed by others
References
Acimovic S, McGhee G, Sundin G, Wise J, Zeng Q (2015) Control of fire blight (Erwinia amylovora) on apple trees with trunk-injected plant resistance inducers and antibiotics and assessment of induction of pathogenesis-related protein genes. Front Plant Sci 6:1–10. https://doi.org/10.3389/fpls.2015.00016
Adaskaveg J, Förster H, Wade M (2010) Effectiveness of Kasugamycin against Erwinia amylovora and its potential use for managing fire blight of pear. Plant Dis 95:448–454. https://doi.org/10.1094/pdis-09-10-0679
Altschul S, Madden T, Schaffer A, Zhang J, Zhang Z, Miller W, Lipman D (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402. https://doi.org/10.1093/nar/25.17.3389
Bahadou S, Ouijja A, Karfach A, Tahiri A, Lahlali R (2018) New potential bacterial antagonists for the biocontrol of fire blight disease (Erwinia amylovora) in Morocco. Microb Pathog 117:7–15. https://doi.org/10.1016/j.micpath.2018.02.011
Balouiri M, Sadiki M, Ibnsouda S (2016) Methods for in vitro evaluating antimicrobial activity: a review. J Pharm Anal 6:71–79. https://doi.org/10.1016/j.jpha.2015.11.005
Beris D, Theologidis I, Skandalis N, Vassilakos N (2018) Bacillus amyloliquefaciens strain MBI600 induces salicylic acid dependent resistance in tomato plants against tomato spotted wilt virus and potato virus y. Sci Rep 8:1–11. https://doi.org/10.1038/s41598-018-28677-3
Broggini G, Duffy B, Holliger E, Scharer H, Gessler C, Patocchi A (2005) Detection of the fire blight biocontrol agent Bacillus subtilis BD170 (biopro (R)) in a Swiss apple orchard. Eur J Plant Pathol 111:93–100. https://doi.org/10.1007/s10658-004-1423-x
Cabrefiga J, Montesinos E (2017) Lysozyme enhances the bactericidal effect of BP100 peptide against Erwinia amylovora, the causal agent of fire blight of rosaceous plants. BMC Microbiol 17:1–10. https://doi.org/10.1186/s12866-017-0957-y
Cabrefiga J, Bonaterra A, Montesinos E (2007) Mechanisms of antagonism of Pseudomonas fluorescens EPS62e against Erwinia amylovora, the causal agent of fire blight. Int Microbiol 10:123–132. https://doi.org/10.2436/20.1501.01.18
Cesbron S, Thomson S, Paulin J, Morel R (2000) Acibenzolar-S-methyl induces the accumulation of defense-related enzymes in apple and protects from fire blight. Eur J Plant Pathol 106:529–536
Chen X, Scholz R, Borriss M, Junge H, Mögel G (2009) Difficidin and bacilysin produced by plant-associated Bacillus amyloliquefaciens are efficient in controlling fire blight disease. J Biotechnol 140:38–44. https://doi.org/10.1016/j.jbiotec.2008.10.015
Daryaei H, Balasubramaniam V, Yousef A, Legan J, Tay A (2015) Lethality enhancement of pressure-assisted thermal processing against Bacillus amyloliquefaciens spores in low-acid media using antimicrobial compounds. Food Control 59:234–242. https://doi.org/10.1016/j.foodcont.2015.05.029
Doolotkeldieva T, Bobusheva S (2016) Fire blight disease caused by Erwinia amylovora on Rosaceae plants in Kyrgyzstan and biological agents to control this disease. Adv Microbiology 6:831–851. https://doi.org/10.4236/aim.2016.611080
Drenova N, Isin MM, Dzhaimurzina AA, Zharmukhamedova GA, Aitkulov AK (2013) Bacterial fire blight in the Republic of Kazakhstan. Plant Health Research and Practice 1:39–48
Duffy B, Holliger E, Walsh F (2014) Streptomycin use in apple orchards did not increase abundance of mobile resistance genes. FEMS Microbiol Lett 350:180–189. https://doi.org/10.1111/1574-6968.12313
Förster H, McGhee G, Sundin G, Adaskaveg J (2015) Characterization of streptomycin resistance in isolates of Erwinia amylovora in California. Phytopathology 105:1302–1310. https://doi.org/10.1094/phyto-03-15-0078-r
Gerami E, Hassanzadeh N, Abdollahi H, Ghasemi A, Heydari A (2013) 1. Evaluation of some bacterial antagonists for biological control of fire blight disease. J. Plant Pathol 95:127–134
Hassan M, Buchenauer H (2016) Induction of resistance to fire blight in apple by acibenzolar-S-methyl and DL-3-aminobutyric acidResistenzinduktion gegenüber Feuerbrand an Apfel durch Acibenzolar-S-methyl und DL-3-Aminobuttersäure. J Plant Dis Prot 114:151–158. https://doi.org/10.1007/bf03356211
Holt J, Krieg N, Sneat P, Staley J, Williams S (1994) Bergey’s manual of determinative bacteriology. Williams & Wilkins, Baltimore
Jock S, Wensing A, Pulawska J, Drenova N, Dreo T, Geider K (2013) Molecular analyses of Erwinia amylovora strains isolated in Russia, Poland, Slovenia and Austria describing further spread of fire blight in Europe. Microbiol Res 168:447–454. https://doi.org/10.1016/j.micres.2013.01.008
Kearns L, Mahanty H (1998) Antibiotic production by Erwinia herbicola Eh1087: its role in inhibition of Erwinia amylovora and partial characterization of antibiotic biosynthesis genes. Appl Environ Microbiol 64:1837–1844
Kumar S, Nei M (2004) MEGA3: integrated software for molecular evolutionary genetics. Brief Bioinform 5:150–163
Lešnik M, Vajs S, Gaberšek V, Kurnik V (2013) Investigation of phytotoxicity regarding copper fungicides applied to apples. Agricultura 1–2:55–59
Mazzucchi A, Brunelli A (2008) Resistance to Erwinia amylovora in immature pears induced by acibenzolar-S-methyl in the orchard. Phytopathol Mediterr 47:272–276
Nischwitz C, Dhiman C (2013) Streptomycin resistance of Erwinia amylovora isolated from apple ( Malus domesticus ) in Utah. Plant Heal Prog 14:1. https://doi.org/10.1094/php-2013-1025-01-rs
Ordax M, Marco-Noales E, López M, Biosca E (2006) Survival strategy of Erwinia amylovora against copper: induction of the viable-but-nonculturable state. Appl Environ Microbiol 72:3482–3488. https://doi.org/10.1128/AEM.72.5.3482-3488.2006
Paternoster T, Défago G, Duffy B, Gessler C, Pertot I (2010) Selection of a biocontrol agent based on a potential mechanism of action: degradation of nicotinic acid, a growth factor essential for Erwinia amylovora. Int Microbiol 13:195–206. https://doi.org/10.2436/20.1501.01.126
Roberts I, Coleman M (2009) The virulence of Erwinia amylovora: molecular genetic perspectives. J Gen Microbiol 137:1453–1457. https://doi.org/10.1099/00221287-137-7-1453
Roselló M, Peñalver J, Llop P, Gorris MT, Cambra M, López M, Chartier R, García F, Montón C (2006) Identification of an Erwinia sp. different from Erwinia amylovora and responsible for necrosis on pear blossoms. Can. J Plant Pathol 28:30–41. https://doi.org/10.1080/07060660609507268
Roselló G, Bonaterra A, Francés J, Montesinos L, Badosa E, Montesinos E (2013) Biological control of fire blight of apple and pear with antagonistic lactobacillus plantarum. Eur J Plant Pathol 137:621–633. https://doi.org/10.1007/s10658-013-0275-7
Sabatini E, Spinelli F, Costa G, Ponti L, Spada G, Geider K, Andreotti C (2015) Chemical control of fire blight in pear: application of Prohexadione-calcium, Acibenzolar-S-methyl, and copper preparations in vitro and under field conditions. Acta Hortic:233–238. https://doi.org/10.17660/actahortic.2006.704.31
Sadanov A, Ismailova E, Iskandarova K, Shemshura O, Seitbattalova A, Molzhigitova A (2018) The structure of the microbial cenosis of phyllosphere of fruit crops affected by fire blight in southern Kazakhstan. Proc. NAS RK. Ser. Biol. Med. 5:71–77. https://doi.org/10.32014/2018.2518-1629.10
Sharifazizi M, Harighi B, Sadeghi A (2017) Evaluation of biological control of Erwinia amylovora, causal agent of fire blight disease of pear by antagonistic bacteria. Biol Control 104:28–34. https://doi.org/10.1016/j.biocontrol.2016.10.007
Shoeib A, Ashmawy N, Hammad S, Hafez E (2017) Molecular and Biological Identification of Erwinia amylovora Egyptian Isolates Compared with Other German Strains. J Plant Physiol Pathol 05. https://doi.org/10.4172/2329-955x.1000156
Soukainen M, Santala J, Tegel J (2015) First report of Erwinia amylovora, the causal agent of fire blight, on pear in Finland. Plant Dis 99:1033–1034. https://doi.org/10.1094/PDIS-11-14-1221-PDN
Tancos K, Cox K (2016) Exploring diversity and origins of streptomycin-resistant Erwinia amylovora isolates in New York through CRISPR spacer arrays. Plant Dis 100:1307–1313. https://doi.org/10.1094/pdis-01-16-0088-re
Turner S, Pryer KM, Miao VPW, Palmer JD (1999) Investigating deep phylogenetic relationships among cyanobacteria and plastids by small subunit rRNA sequence analysis. J Eukaryot Microbiol 46:327–338
Wisniewski M, Evans K, Norelli J, Harshman J, Aldwinckle H, Flamenco J, Potts R, Allen H (2017) Fire Blight Resistance in Wild Accessions of Malus sieversii . Plant Dis. 101, PDIS-01-17-0077. doi:https://doi.org/10.1094/pdis-01-17-0077-re
Woese C, Kandlert O, Wheelis M (1990) Towards a natural system of organisms: proposal for the domains Archaea, Bacteria, and Eucarya (Euryarchaeota/Crenarchaeota/kingdom/evolution). Proc Nati Acad Sci USA 87:4576–4579. https://doi.org/10.1136/bmj.1.4540.46
Yashiro E, McManus P (2012) Effect of streptomycin treatment on bacterial community structure in the apple phyllosphere. PLoS One 7. https://doi.org/10.1371/journal.pone.0037131
Yuan B, Wang Z, Qin S, Zhao G-H, Feng Y-J, Wei L-H, Jiang J-H (2012) Study of the anti-sapstain fungus activity of Bacillus amyloliquefaciens CGMCC 5569 associated with Ginkgo biloba and identification of its active components. Bioresour Technol 114:536–541. https://doi.org/10.1016/j.biortech.2012.03.062
Zengerer V, Bieri M, Remus-Emsermann M, Müller D, Pelludat C, Schmid M, Ahrens C (2018) Pseudomonas orientalis F9: a potent antagonist against Phytopathogens with phytotoxic effect in the apple flower. Front Microbiol 9:1–13. https://doi.org/10.3389/fmicb.2018.00145
Zhang S, Wang Y, Meng L, Li J, Zhao X, Cao X, Chen X, Wang A, Li J (2012) Isolation and characterization of antifungal lipopeptides produced by endophytic Bacillus amyloliquefaciens TF28. African J Microbiol Res 6:1747–1755. https://doi.org/10.5897/ajmr11.1025
Zhang Y, Li S, Liu L, Wu J (2013) Acetoin production enhanced by manipulating carbon flux in a newly isolated Bacillus amyloliquefaciens. Bioresour Technol 130:256–260. https://doi.org/10.1016/j.biortech.2012.10.036
Acknowledgments
This study was funded bythe Ministry of Education and Science of the Republic of Kazakhstan (Grant no. AP0131543). Author Amankeldy Sadanov has received research grants from Ministry of Education and Science of the Republic of Kazakhstan. The authors declare that they have no conflict of interest. This article does not contain any studies with human participants or animals performed by any of the authors. The authors are grateful to Dr. Lars Carlsen (Awareness Center, Denmark) for assistance in preparation of the final version of the paper.
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Shemshura, O., Alimzhanova, M., Ismailova, E. et al. Antagonistic activity and mechanism of a novel Bacillus amyloliquefaciens MB40 strain against fire blight. J Plant Pathol 102, 825–833 (2020). https://doi.org/10.1007/s42161-020-00515-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s42161-020-00515-4