Effect of antimicrobial peptides and monoterpenes on control of fire blight

  • Mahdi Akhlaghi Ferdowsi University of Mashhad, Laboratory of Phytopathology, Dept. Crop Protection, Mashhad
  • Saeed Tarighi Ferdowsi University of Mashhad, Laboratory of Phytopathology, Dept. Crop Protection, Mashhad
  • Parissa Taheri Ferdowsi University of Mashhad, Laboratory of Phytopathology, Dept. Crop Protection, Mashhad
Keywords: antibacterial, biological control, central composite design, Erwinia amylovora, guaiacol peroxidase, induced resistance


Aim of study: Antimicrobial peptides and monoterpenes are safe compounds that have been used for control of many plant diseases. Herein, the effects of two recombinant antibacterial peptides (AMPs) were compared with two monoterpenes for control of Erwinia amylovora directly or via induction of plant defense enzyme guaiacol peroxidase (GPOD).

Area of study: The experiments were performed at the Ferdowsi University of Mashhad (Iran).

Material and methods: The central composite design (CCD) method was used to study the effect of mixing the compounds and copper compound (Nordox) in controlling the pathogen. The resistance level was studied on shoots of tolerant (‘Dargazi’) and semi-susceptible (‘Spadona’) pear cultivars treated with the antibacterial compounds.

Main results: Thanatin and 1,8-cineole showed the highest and lowest antibacterial effects. All treatments reduced E. amylovora pathogenicity on blossom. The CCD analysis revealed that the best reduction in colony number obtained by mixing Lfc, thanatin, thymol, 1,8-cineole and Nordox at concentrations of 32, 16, 24, 250 and 250 μg/mL. Thymol and 1,8-cineole at 500 μg/mL decreased disease severity significantly compared to that of AMPs. The level of GPOD enzyme in ‘Dargazi’ was higher than in ‘Spadona’. All treatments increased the GPOD levels in both cultivars. Furthermore, resistance level and GPOD ratio were negatively correlated.

Research highlights: Antimicrobial peptides showed better effect on growth inhibition of E. amylovora than monoterpenes. Mixing of these peptides and monoterpens at special dosage enhanced their antimicrobial efficacy against E. amylovora; that could represent a new method in control of fire blight disease.


Download data is not yet available.


Akhlaghi M, Tarighi S, Taheri P, 2018. Evaluating antibacterial effect of plant extracts against Erwinia amylovora and their role in resistance induction in pear. BCPPD 7 (2): 31-47.

Al-Daoude, A, Arabi M, Ammouneh H, 2009. Studying Erwinia amylovora isolates from Syria for copper resistance and streptomycin sensitivity. J Plant Pathol 91: 203-205.

Awais Khan M, Zhao Y, Korban SS, 2011. Molecular mechanisms of pathogenesis and resistance to the bacterial pathogen Erwinia amylovora, causal agent of fire blight disease in Rosaceae. Plant Mol Biol Rep 30: 247-260. https://doi.org/10.1007/s11105-011-0334-1

Bassolé IHN, Juliani HR, 2012. Essential oils in combination and their antimicrobial properties. Molecules 17: 3989-4006. https://doi.org/10.3390/molecules17043989

Bell AC, Ranney TG, Eaker TA, 2004. Resistance to fire blight among flowering pear and quince. Hortscience 40 (2): 413-415. https://doi.org/10.21273/HORTSCI.40.2.413

Bellemann P, Bereswill S, Berger S, Geider K, 1994. Visualization of capsule formation by Erwinia amylovora and assays to determine amylovoran synthesis. Int J Biol Macromol 16: 290-296. https://doi.org/10.1016/0141-8130(94)90058-2

Ben Kaab S, Rebey IB, Hanafi M, Berhal C, Fauconnier ML, de Clerck C, Ksouri R, Jijakli H, 2019. Rosmarinus officinalis essential oil as an effective antifungal and herbicidal agent. Span J Agric Res 17 (2): 1-9. https://doi.org/10.5424/sjar/2019172-14043

Biswaro LS, Da Costa Sousa MG, Rezende TMB, Dias SC, Franco OL, 2018. Antimicrobial peptides and nanotechnology, recent advances and challenges. Front Microbiol 9: 855. https://doi.org/10.3389/fmicb.2018.00855

Bolscher JG, Adão R, Nazmi K, van den Keybus PA, van't Hof W, Nieuw Amerongen AV, Bastos M, Veerman EC, 2009. Bactericidal activity of LF chimera is stronger and less sensitive to ionic strength than its constituent lactoferricin and lactoferrampin peptides. Biochimie 91 (1): 123-132. https://doi.org/10.1016/j.biochi.2008.05.019

Brown SK, Garver, WS, Orlando RA, 2017. 1,8-cineole: An underappreciated anti-inflammatory therapeutic. J Biomol Res Ther 6 (154): 1-6. https://doi.org/10.4172/2167-7956.1000154

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 (39): 1-10. https://doi.org/10.1186/s12866-017-0957-y

Calvet C, Pinochet J, Camprubi A, Estaun V, Rodriguez-Kabana R, 2001. Evaluation of natural chemical compounds against root lesion and root-knot nematodes and side-effects on the infectivity of arbuscular mycorrhizal fungi. Eur J Plant Pathol 107: 601-605. https://doi.org/10.1023/A:1017954315942

Chouhan S, Sharma K, Guleria S, 2017. Antimicrobial activity of some essential oils present status and future perspectives. Medicines 4 (58): 1-21. https://doi.org/10.3390/medicines4030058

Demirel M, Kayan B, 2012. Application of response surface methodology and central composite design for the optimization of textile dye degradation by wet air oxidation. Int J Ind Chem 3 (24): 1-10. https://doi.org/10.1186/2228-5547-3-24

Ebadi A, Erfani J, Abdollahi H, Fattahi Moghaddam J, 2014. Investigation of changes in antioxidant enzyme and total phenol level in some pear cultivars inoculated with fire blight disease. Ir J Hortic Sci 45 (2): 127-136.

Emeriewen OF, Wöhner T, Flachowsky H, Peil A, 2019. Malus hosts-Erwinia amylovora interactions: strain pathogenicity and resistance mechanisms. Front Plant Sci 10 (551): 1-7. https://doi.org/10.3389/fpls.2019.00551

Fischer TC, Gosch C, Mirbeth B, Gselmann M, Thallmair V, Stich K, 2012. Potent and specific bactericidal effect of juglone (5-hydroxy-1, 4-naphthoquinone) on the fire blight pathogen Erwinia amylovora. J Agr Food Chem 60: 12074-12081. https://doi.org/10.1021/jf303584r

Fukuta S, Kawamoto KI, Mizukami Y, Yoshimura Y, Ueda JI, Kanbe M, 2012. Transgenic tobacco plants expressing antimicrobial peptide bovine lactoferricin show enhanced resistance to phytopathogens. Plant Biotechnol 29: 383-389. https://doi.org/10.5511/plantbiotechnology.12.0619a

Gerami E, Hassanzadeh N, Abdollahi H, Ghasemi A, Heydari A, 2013. Evaluation of some bacterial antagonists for biological control of fire blight disease. J Plant Pathol 95 (1): 127-134.

Gusberti M, Klemm U, Meier MS, Maurhofer M, Hunger-Glaser I, 2015. Fire blight control: The struggle goes on. A comparison of different fire blight control methods in Switzerland with respect to biosafety, efficacy and durability. Int J Environ Res Public Health 12: 11422-11447. https://doi.org/10.3390/ijerph120911422

Hassanzadeh N, 2005. The essential oil of thyme as a natural plant extract for fire blight control. Proc 1 Int Conf on biological control of bacterial plant diseases, Seeheim/Darmstadt (Germany), Oct 23-26. pp: 290-292.

Holaskova E, Galuszka P, Frebort I, Oz MT, 2015. Antimicrobial peptide production and plant-based expression systems for medical and agricultural biotechnology. Biotechnol Adv 33: 1005-1023. https://doi.org/10.1016/j.biotechadv.2015.03.007

Iacobellis NS, Lo Cantore P, Capasso F, Senatore F, 2005. Antibacterial activity of Cuminum cyminum L. and Carum carvi L. essential oils. J Agric Food Chem 53: 57-61. https://doi.org/10.1021/jf0487351

Imamura T, Yasuda M, Kusano H, Nakashita H, Ohno Y, Kamakura T, Taguchi S, Shimada H, 2010. Acquired resistance to the rice blast in transgenic rice accumulating the antimicrobial peptide thanatin. Transgenic Res 19: 415-424. https://doi.org/10.1007/s11248-009-9320-x

Ivanović M, Gasić K, Prokić A, Kuzmanović N, Zlatković N, Obradović A, 2016. Screening for copper and antibiotic resistance in Erwinia amylovora population from Serbia. Acta Hortic 1139: 715-720. https://doi.org/10.17660/ActaHortic.2016.1139.122

Ji P, Momol, MT, Olson SM, Pradhanang PM, Jones JB, 2005. Evaluation of thymol as bio fumigant for control of bacterial wilt of tomato under field conditions. Plant Dis 89: 497-500. https://doi.org/10.1094/PD-89-0497

Johnson KB, Temple TN, 2013. Evaluation of strategies for fire blight control in organic pome fruit without antibiotics. Plant Dis 97: 402-409. https://doi.org/10.1094/PDIS-07-12-0638-RE

Kar M, Mishra D, 1976. Catalase, peroxidase and polyphenol oxidase activities during rice leaf senescence. Plant Physiol 57: 315-319. https://doi.org/10.1104/pp.57.2.315

Karami-Osboo R, Khodaverdi M, Ali-Akbari F, 2010. Antibacterial effect of effective compounds of Satureja hortensis and Thymus vulgaris essential oils against Erwinia amylovora. J Agr Sci Tech 12: 35-45.

Koczan JM, McGrath MJ, Zhao Y, Sundin GW, 2009. Contribution of Erwinia amylovora exopolysaccharides amylovoran and levan to biofilm formation: implications in pathogenicity. Phytopathol 99: 1237-1244. https://doi.org/10.1094/PHYTO-99-11-1237

Koul O, Walia S, Dhaliwal GS, 2008. Essential oil as green pesticides: Potential and constraints. Biopestic Int 4: 63-84.

Langeveld WT, Veldhuizen EJA, Burt SA, 2014. Synergy between essential oil components and antibiotics: a review. Crit Rev Microbiol 40 (1): 76-94. https://doi.org/10.3109/1040841X.2013.763219

Lelliot RA, Stead DE, 1987. Methods for the diagnosis of bacterial diseases of plants. In: Methods in plant pathology; Preece TF (eds.). pp. 1-216. Blackwell Sci Publ, Oxford, UK.

Li Y, 2011. Recombinant production of antimicrobial peptides in Escherichia coli: A review. Protein Expr Purif 80 (2): 260-267. https://doi.org/10.1016/j.pep.2011.08.001

Mamarabadi M, Tanhaeian A, Ramezany Y, 2018. Antifungal activity of recombinant thanatin in comparison with two plant extracts and a chemical mixture to control fungal plant pathogens. AMB Expr 8 (180): 1-12. https://doi.org/10.1186/s13568-018-0710-4

Mandal S, Mitra A, Mallick, N, 2008. Biochemical characterization of oxidative burst during interaction between Solanum lycopersicum and Fusarium oxysporum f. sp. Lycopersici. Physiol Mol Plant Pathol 72: 56-61. https://doi.org/10.1016/j.pmpp.2008.04.002

Mandard N, Sodano P, Labbe H, Bonmatin JM, Bulet P, Hetru C, Ptak M, Vovelle F, 1998. Solution structure of thanatin, a potent bactericidal and fungicidal insect peptide, determined from proton two-dimensional nuclear magnetic resonance data. Eur J Biochem 256: 404-410. https://doi.org/10.1046/j.1432-1327.1998.2560404.x

Mansfield J, Genin S, Magori S, Citovsky V, Sriariyanum M, Ronald P, Dow M, Verdier V, Beer SV, Machado MA, et al., 2012. Top 10 plant pathogenic bacteria in molecular plant pathology. Mol Plant Pathol 13: 614-629. https://doi.org/10.1111/j.1364-3703.2012.00804.x

Marcos JF, Munoz A, Perez-Paya E, Misra S, Lopez-Garcıa B, 2008. Identification and rational design of novel antimicrobial peptides for plant protection. Annu Rev Phytopathol 46: 273-301. https://doi.org/10.1146/annurev.phyto.121307.094843

Mirzaei-Najafgholi H, Tarighi S, Golmohammadi M, Taheri P, 2017. The effect of citrus essential oils and their constituents on growth of Xanthomonas citri subsp. citri. Molecules 22 (591): 2-14. https://doi.org/10.3390/molecules22040591

Mittler R, 2002. Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7: 405-410. https://doi.org/10.1016/S1360-1385(02)02312-9

Ozrenk K, Balta F, Celik F, 2011. Levels of fire blight (Erwinia amylovora) susceptibility of native apple, pear and quince germplasm from Lake Van Basin, Turkey. Eur J Plant Pathol 132 (2): 229-236. https://doi.org/10.1007/s10658-011-9866-3

Patel RR, Sundin GW, Yang CH, Wang J, Huntley RB, Yuan X, Zeng Q, 2017. Exploration of using antisense peptide nucleic acid (PNA)-cell penetrating peptide (CPP) as a novel bactericide against fire blight pathogen Erwinia amylovora. Front Microbiol 8 (687): 1-12. https://doi.org/10.3389/fmicb.2017.00687

Piqué N, Miñana-Galbis D, Merino S, Tomás JM, 2015. Virulence factors of Erwinia amylovora: A review. Int J Mol Sci 16: 12836-12854. https://doi.org/10.3390/ijms160612836

Plewa MJ, Smith SR, Wanger ED, 1991. Diethyldithiocarbamate suppresses the plant activation of aromatic amines into mutagens by inhabitation tobacco cell peroxidase. Mutat Res 247 (1): 57-64. https://doi.org/10.1016/0027-5107(91)90033-K

Salehi B, Mishra AP, Shukla I, Sharifi‐Rad M, Contreras MDM, Segura‐Carretero A, Fathi H, Nasri Nasrabadi N, Kobarfard F, Sharifi‐Rad J, 2017. Thymol, thyme, and other plant sources: health and potential Uses. Phytother Res 32 (9): 1688-1706. https://doi.org/10.1002/ptr.6109

Skłodowska M, Gajewska E, Kuźniak E, Wielanek M, Mikicinski A, Sobiczewski P, 2011. Antioxidant profile and polyphenol oxidase activities in apple leaves after Erwinia amylovora infection and pretreatment with a benzothiadiazole-type resistance inducer (BTH). J Phytopathol 159: 495-504. https://doi.org/10.1111/j.1439-0434.2011.01793.x

Soares V, Rodrigues FB, Vieira MF, Silva MS, 2011. Validation of a protocol to evaluate maximal expiratory pressure using a pressure transducer and a signal conditioner. An Acad Bras Cienc 83: 967-971. https://doi.org/10.1590/S0001-37652011005000021

Sokovic M, Van Griensven L, 2006. Antimicrobial activity of essential oils and their components against the three major pathogens of the cultivated button mushroom, Agaricus bisporus. Eur J Plant Pathol 116: 211-224. https://doi.org/10.1007/s10658-006-9053-0

Somayajula A, Asaithambi P, Susree M, Matheswaran M, 2011. Sonoelectrochemical oxidation for decolorization of reactive red 195. Ultrason Sonochem 19: 803-811. https://doi.org/10.1016/j.ultsonch.2011.12.019

Steiner P, 2000. Integrated orchard and nursery management for the control of fire blight. In: Fire blight: The disease and its causative agent, Erwinia amylovora; Vanneste JL (eds.). pp. 339-358. CABI Publ, NY, USA. https://doi.org/10.1079/9780851992945.0339

Sticher L, MauchMani B, Metraux, JP, 1997. Systemic acquired resistance. Ann Rev Phytopathol 35: 235-270. https://doi.org/10.1146/annurev.phyto.35.1.235

Tang XS, Tang ZR, Wang SP, Feng ZM, Zhou D, Li TJ, Yin YL, 2012. Expression, purification, and antibacterial activity of bovine Lactoferrampin-Lactoferricin in Pichia pastoris. Appl Biochem Biotechnol 166 (3): 640-651. https://doi.org/10.1007/s12010-011-9455-0

Tanhaeian A, Shahriari Ahmadi F, Sekhavati MH, Mamarabadi M, 2018. Expression and purification of the main component contained in camel milk and its antimicrobial activities against bacterial plant pathogens. Probiotics Antimicrob Proteins 10 (4): 787-793. https://doi.org/10.1007/s12602-018-9416-9

Tanhaeian A, 2018. Production of the recombinant chimeric peptide, Lactoferrampin-Lactoferricin of camel milk in different expression systems and its bioactivity evaluation. Doctoral thesis. Univ. Ferdowsi, Mashhad, Iran.

Thomson SV, 1986. The role of the stigma in fire blight infections. Phytopathol 76: 476-482. https://doi.org/10.1094/Phyto-76-476

Van Vuuren SF, Viljoen AM, 2007. Antimicrobial activity of limonene enantiomers and 1,8-Cineole alone and in combination. Flavour Fragr J 22: 540-544. https://doi.org/10.1002/ffj.1843

Viljevac M, Dugali K, Stolfa I, Dermi E, Cvjetkovi B, Sudar R, Kovacevi J, Cesar V, Lepedus H, Jurkovic Z, 2009. Biochemical basis of apple leaf resistance to Erwinia amylovora infection. Food Technol Biotechnol 47 (3): 281-287.

Wang WY, Wong JH, Ip DTM, Wan DCC, Cheung RC, Ng TB, 2016. Bovine lactoferrampin, human lactoferricin, and lactoferrin 1-11 inhibit nuclear translocation of HIV integrase. Appl Biochem Biotechnol 179 (7): 1202-1212. https://doi.org/10.1007/s12010-016-2059-y

Winslow CEA, Broadhurst J, Buchanan RE, Krumwiede C, Rogers LA, Smith GH, 1920. The families and genera of the bacteria. Final report of the committee of the society of American bacteriologists on characterization and classification of bacterial types. J Bacteriol 5: 191-229. https://doi.org/10.1128/JB.5.3.191-229.1920

Wu T, Tang D, Chen W, Huang H, Wang R, Chen Y, 2013. Expression of antimicrobial peptides thanatin (S) in transgenic Arabidopsis enhanced resistance to phytopathogenic fungi and bacteria. Gene 527: 235-242. https://doi.org/10.1016/j.gene.2013.06.037

Yumlembam RA, Borkar SG. 2014. Assessment of antibacterial properties of medicinal plants having bacterial leaf endophytes against plant pathogenic Xanthomonads. Ind Phytopath 67 (4): 353-357.

Zhang Z, Zheng H, 2009. Optimization for decolorization of azo dye acid green 20 by ultrasound and H2O2 using response surface methodology. J Hazard Mater 172: 1388-1393. https://doi.org/10.1016/j.jhazmat.2009.07.146

Zhang Z, Nakano K, Maezawa S, 2009. Comparison of the antioxidant enzymes of broccoli after cold or heat shock treatment at different storage temperatures. Postharvest Biol Technol 54: 101-105. https://doi.org/10.1016/j.postharvbio.2009.05.006

How to Cite
Akhlaghi, M., Tarighi, S., & Taheri, P. (2020). Effect of antimicrobial peptides and monoterpenes on control of fire blight. Spanish Journal of Agricultural Research, 18(2), e1002. https://doi.org/10.5424/sjar/2020182-15629
Plant protection