scholarly journals Evaluation of Three Antimicrobial Peptides Mixtures to Control the Phytopathogen Responsible for Fire Blight Disease

Plants ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 2637
Author(s):  
Rafael Mendes ◽  
Sara Sario ◽  
João Pedro Luz ◽  
Natália Tassi ◽  
Cátia Teixeira ◽  
...  
Keyword(s):  
Antimicrobial Peptides ◽  
Fire Blight ◽  
Green Revolution ◽  
Fruit Trees ◽  
Control Measures ◽  
Phytopathogenic Bacterium ◽  
Viable Cells ◽  
Efficient Control ◽  
High Reduction ◽  
Blight Disease

Fire blight is a severe bacterial plant disease that affects important chain-of-value fruit trees such as pear and apple trees. This disease is caused by Erwinia amylovora, a quarantine phytopathogenic bacterium, which, although highly distributed worldwide, still lacks efficient control measures. The green revolution paradigm demands sustainable agriculture practices, for which antimicrobial peptides (AMPs) have recently caught much attention. The goal of this work was to disclose the bioactivity of three peptides mixtures (BP100:RW-BP100, BP100:CA-M, and RW-BP100:CA-M), against three strains of E. amylovora representing distinct genotypes and virulence (LMG 2024, Ea 630 and Ea 680). The three AMPs’ mixtures were assayed at eight different equimolar concentrations ranging from 0.25 to 6 μM (1:1). Results showed MIC and MBC values between 2.5 and 4 μM for every AMP mixture and strain. Regarding cell viability, flow cytometry and alamarBlue reduction, showed high reduction (>25%) of viable cells after 30 min of AMP exposure, depending on the peptide mixture and strain assayed. Hypersensitive response in tobacco plants showed that the most efficient AMPs mixtures and concentrations caused low to no reaction of the plant. Altogether, the AMPs mixtures studied are better treatment solutions to control fire blight disease than the same AMPs applied individually.

scholarly journals In Vitro Evaluation of Five Antimicrobial Peptides against the Plant Pathogen Erwinia amylovora

Biomolecules ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 554
Author(s):  
Rafael J. Mendes ◽  
Laura Regalado ◽  
João P. Luz ◽  
Natália Tassi ◽  
Cátia Teixeira ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Antimicrobial Peptides ◽  
Erwinia Amylovora ◽  
Growth Curves ◽  
Fruit Trees ◽  
Membrane Permeabilization ◽  
Control Measures ◽  
Minimal Bactericidal Concentration ◽  
Pome Fruit

Fire blight is a major pome fruit trees disease that is caused by the quarantine phytopathogenic Erwinia amylovora, leading to major losses, namely, in pear and apple productions. Nevertheless, no effective sustainable control treatments and measures have yet been disclosed. In that regard, antimicrobial peptides (AMPs) have been proposed as an alternative biomolecule against pathogens but some of those AMPs have yet to be tested against E. amylovora. In this study, the potential of five AMPs (RW-BP100, CA-M, 3.1, D4E1, and Dhvar-5) together with BP100, were assessed to control E. amylovora. Antibiograms, minimal inhibitory, and bactericidal concentrations (minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC), growth and IC50 were determined and membrane permeabilization capacity was evaluated by flow cytometry analysis and colony-forming units (CFUs) plate counting. For the tested AMPs, the higher inhibitory and bactericidal capacity was observed for RW-BP100 and CA-M (5 and 5–8 µM, respectively for both MIC and MBC), whilst for IC50 RW-BP100 presented higher efficiency (2.8 to 3.5 µM). Growth curves for the first concentrations bellow MIC showed that these AMPs delayed E. amylovora growth. Flow cytometry disclosed faster membrane permeabilization for CA-M. These results highlight the potential of RW-BP100 and CA-M AMPs as sustainable control measures against E. amylovora.

scholarly journals Effect of antimicrobial peptides and monoterpenes on control of fire blight

2020 ◽  
Vol 18 (2) ◽  
pp. e1002
Author(s):  
Mahdi Akhlaghi ◽  
Saeed Tarighi ◽  
Parissa Taheri
Keyword(s):  
Antimicrobial Peptides ◽  
Fire Blight ◽  
Erwinia Amylovora ◽  
Plant Diseases ◽  
Guaiacol Peroxidase ◽  
Resistance Level ◽  
Defense Enzyme ◽  
Colony Number ◽  
Blight Disease ◽  
Central Composite

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.

scholarly journals Causative Pathogen of Fire Blight of Fruit Trees in Georgia

2021 ◽  
Author(s):  
Nanuli Amashukeli ◽  
Dali Gaganidze ◽  
Mariam Aznarashvili ◽  
Shorena Kharadze ◽  
Neli Sturua ◽  
...  
Keyword(s):  
Fire Blight ◽  
Erwinia Amylovora ◽  
Fruit Trees ◽  
Visual Observation ◽  
Pcr Analysis ◽  
Causative Pathogen ◽  
Plant Samples ◽  
Immunological Test ◽  
Blight Disease

Fire blight is a devastating disease of fruit trees that first appeared in Georgia in 2016. Above 40 samples – fruit trees twigs, buds, flowers and fruitlets, suspected on the fire blight disease were collected in the Mtskheta-Mtianeti and Shida Kartli Regions, eastern Georgia in Summer of 2020. Based on visual observation and immunological test (Ea AgriStrip), 20 plant samples were selected for further study. PCR analysis was performed to detect pathogen in these samples with primers pair A: 5′-CGG TTT TTA ACG CTG GG-3′ and B: 5′-GGG CAA ATA CTC GGA TT-3 ′ and 2XPCR BIOTAG Mix using PCR BIOSYSTEMS. The pathogen Erwinia amylovora was detected in 6 samples: N10 and N11 apple samples (Ksovrisi);

scholarly journals Effectiveness of a Peptide-rich Fraction from Xenorhabdus budapestensis Culture against Fire Blight Disease on Apple Blossoms

2015 ◽  
Vol 43 (2) ◽  
pp. 547-553 ◽  
Author(s):  
Dávid VOZIK ◽  
Katalin BÉLAFI-BAKÓ ◽  
Mária HEVESI ◽  
Erzsébet BÖSZÖRMÉNYI ◽  
András FODOR
Keyword(s):  
Fire Blight ◽  
Erwinia Amylovora ◽  
Control Method ◽  
Plant Protection ◽  
Test Method ◽  
In Planta ◽  
Phytopathogenic Bacterium ◽  
Protection Potential ◽  
Blight Disease

Erwinia amylovora is one of the most frequently occurred plant pathogenic bacterium. It causes necrosis and blight symptoms on host plantsand it lead to considerable yield losses throughout the world on apple trees. There is no effective chemical treatment is currently available against fire blight. The purpose of the present study was to search a new, alternative control method. The evaluation of the plant protection potential of an enriched fraction of Xenorhabdus budapestensis cell-free conditioned media was investigated. Purified samples were tested in vitro and in plantaagainst the phytopathogenic bacterium. A reproducible method for isolation of a peptide-rich fraction from Xenorhabdus cell-free conditionedmedia was established. The process resulted in 400 mg of dry sample prepared from three litres of Xenorhabdus cell culture. Significant correlation was found between the concentration of the purified preparation and the induced inactivation zones against Erwinia amylovora Ea1 in agardiffusion test method. The minimum inhibitory concentration and minimum bactericidal concentration of the purified fraction against Ea1strain were 8 ;g/mL and 16 ;g/mL, respectively. In planta experiments were tested on an apple cultivar (‘Watson Jonathan’) susceptible to fireblight. The effective range of concentration was 62-200 μg/mL, while treatment with 300 μg/mL and larger amounts caused necrotic symptomson the petals of flowers. Current study pointed to the effectiveness of the compounds produced by X. budapestensis against fire blight. Thedevelopment of a commercially applicable formulation of these compounds would allow growers to effectively control fire blight in apple and pearorchards.

scholarly journals Evaluation of host resistance inducers and conventional products for fire blight management in loquat and quince

2008 ◽  
Vol 88 (3) ◽  
pp. 93-101 ◽  
Author(s):  
Kubilay Kurtulus Bastas ◽  
Salih Maden
Keyword(s):  
Humic Acid ◽  
Disease Severity ◽  
Fire Blight ◽  
Fruit Trees ◽  
Pome Fruit ◽  
Copper Salts ◽  
Harpin Protein ◽  
Shoot Blight ◽  
Resistance Inducers ◽  
Blight Disease

Abstract Fire blight disease is one of the most destructive diseases of pome fruits. Due to the lack of effective, non-phytotoxic and publicly acceptable materials for controlling fire blight in pome fruit trees, new strategies to manage Erwinia amylovora fire blight are being sought. The resistance-inducing compounds prohexadione-Ca, harpin protein and benzothiadiazole (acibenzolar-S-methyl), the fertilizer humic acid, the bactericides streptomycin and copper salts, and combinations of copper with chemicals were evaluated for their ability to control fire blight on quince and loquat cultivars. Prohexadione-Ca was applied at a rate of 125 mg L-1 at two shoot lengths (6-12 cm and 15-20 cm), while benzothiadiazole + metalaxyl (135 mg L-1) and harpin (50 mg L-1) were applied when the shoots measured between 15-20 cm, and again at 30-35 cm. On loquat cv. Cukurgobek, benzothiadiazole + metalaxyl showed about 60% effectiveness. The addition of copper salts reduced the effectiveness of benzothiadiazole + metalaxyl. On quince cultivars, streptomycin (P ≤ 0.05) was the most effective treatment during both years, followed by the harpin protein alone and in combination with copper salts. Prohexadione-Ca, benzothiadiazole + metalaxyl, and harpin protein applications reduced disease severity on inoculated shoots compared with copper and untreated controls. Prohexadione-Ca reduced both shoot length and shoot blight on the two hosts. Humic acid applications were ineffective in controlling fire blight on loquat and quince cultivars. Quince cv. Eşme showed lower disease severity than cv. Ekmek (P ≤ 0.05). The use of resistance-inducing substances during the early phase of shoot growth may offer a means of managing the shoot blight phase of fire blight disease on quince and loquat.

Characteristics of Erwinia amylovora bacteriophage and its possible role in the epidemiology of fire blight

1973 ◽  
Vol 19 (7) ◽  
pp. 837-845 ◽  
Author(s):  
J. M. Erskine
Keyword(s):  
Uv Irradiation ◽  
Fire Blight ◽  
Erwinia Amylovora ◽  
Resistant Mutant ◽  
Fruit Trees ◽  
Symptom Development ◽  
Phage Resistant Mutant ◽  
Pathogenicity Tests ◽  
Optimal Efficiency ◽  
Blight Disease

Bacteriophage (S1) of Erwinia amylovora, isolated from soil at the base of fire-blight-infected trees, was characterized by small, clear plaques on E. amylovora strain PR1 and hazy plaques of the same size on a closely related, yellow, saprophytic bacterium, Y, which was isolated together with PR1 from the diseased trees. Phage S1 plated with optimal efficiency at 10C on PR1 and at 28C on Y was relatively unstable to storage at temperatures typical of summer and to ultraviolet (UV) irradiation, and lysogenized Y but not PR1. Pathogenicity tests in pear slices demonstrated that symptom development was (i) delayed when mixtures of either PR1 and phage S1 or PR1 and Y were inoculated, (ii) delayed when a culture of a phage-resistant mutant of PR1 was inoculated, and (iii) prevented from appearing at all when PR1 and the lysogenic form of Y, Y (S1), were inoculated together. Strain Y was easily lysogenized and Y (S1) released phage spontaneously and after UV irradiation. It is suggested that the yellow saprophyte, which is invariably isolated from fruit trees with E. amylovora, may frequently occur in its lysogenic form in nature and serve as a reservoir of phage which may exert some influence on the occurrence and severity of fire-blight disease.

scholarly journals Virulence Genetics of an Erwinia amylovora Putative Polysaccharide Transporter Family Member

2020 ◽  
Vol 202 (22) ◽  
Author(s):  
Sara M. Klee ◽  
Judith P. Sinn ◽  
Elena Christian ◽  
Aleah C. Holmes ◽  
Kaixi Zhao ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Fire Blight ◽  
Erwinia Amylovora ◽  
Control Measures ◽  
Detectable Effect ◽  
Wild Type ◽  
Content Type ◽  
Transporter Family ◽  
Blight Disease ◽  
Suppressor Screen

ABSTRACT The Gram-negative enterobacterium Erwinia amylovora causes fire blight disease in apple and pear trees. Lipopolysaccharides and the exopolysaccharide amylovoran are essential E. amylovora virulence factors. We found that mutations in rfbX disrupted amylovoran production and virulence in apple fruits and tree shoots and that the deletion of yibD suppressed the rfbX mutant phenotype. The level of expression of yibD was about 10-fold higher in the ΔrfbX mutant than the wild type. A forward genetic suppressor screen in the ΔrfbX mutant uncovered multiple mutations in yibD and supported the conclusion that the virulence defect of rfbX mutants is due to reduced amylovoran production. The yibD and rfbX genes are expressed as a two-gene operon, yibD rfbX. The rfbX gene encodes a previously uncharacterized putative polysaccharide subunit transporter, while yibD encodes a predicted glycosyltransferase. Mutation of rfbX did not have a detectable effect on lipopolysaccharide patterns; however, the overexpression of yibD in both the wild-type and ΔyibD ΔrfbX genetic backgrounds disrupted both amylovoran and lipopolysaccharide production. Additionally, the overexpression of yibD in the ΔyibD ΔrfbX mutant inhibited bacterial growth in amylovoran-inducing medium. This growth inhibition phenotype was used in a forward genetic suppressor screen and reverse-genetics tests to identify several genes involved in lipopolysaccharide production, which, when mutated, restored the ability of the ΔyibD ΔrfbX mutant overexpressing yibD to grow in amylovoran-inducing medium. Remarkably, all the lipopolysaccharide gene mutants tested were defective in lipopolysaccharide and amylovoran production. These results reveal a genetic connection between amylovoran and lipopolysaccharide production in E. amylovora. IMPORTANCE This study discovered previously unknown genetic connections between exopolysaccharide and lipopolysaccharide production in the fire blight pathogen Erwinia amylovora. This represents a step forward in our understanding of the biology underlying the production of these two macromolecules. Fire blight is an economically important disease that impacts the production of apples and pears worldwide. Few fire blight control measures are available, and growers rely heavily on antibiotic applications at bloom time. Both exopolysaccharide and lipopolysaccharide are E. amylovora virulence factors. Our results indicate that the overexpression of the yibD gene in E. amylovora disrupts both lipopolysaccharide production and exopolysaccharide production. This effect could potentially be used as the basis for the development of an antivirulence treatment for the prevention of fire blight disease.

Isolation and characterization of Hena1 – a novel Erwinia amylovora bacteriophage

2020 ◽  
Vol 367 (9) ◽  
Author(s):  
Natalya V Besarab ◽  
Artur E Akhremchuk ◽  
Maryna A Zlatohurska ◽  
Liudmyla V Romaniuk ◽  
Leonid N Valentovich ◽  
...  
Keyword(s):  
Pathogenic Bacteria ◽  
Fire Blight ◽  
Antimicrobial Agents ◽  
Erwinia Amylovora ◽  
Gc Content ◽  
Control Measures ◽  
Effective Control ◽  
Lytic Phage ◽  
Trna Genes ◽  
Isolation And Characterization

ABSTRACT Fire blight, caused by plant pathogenic bacterium Erwinia amylovora, is one of the most important diseases of Rosaceae plants. Due to the lack of effective control measures, fire blight infections pose a recurrent threat on agricultural production worldwide. Recently, bacterial viruses, or bacteriophages, have been proposed as environmentally friendly natural antimicrobial agents for fire blight control. Here, we isolated a novel bacteriophage Hena1 with activity against E. amylovora. Further analysis revealed that Hena1 is a narrow-host-range lytic phage belonging to Myoviridae family. Its genome consists of a linear 148,842 bp dsDNA (48.42% GC content) encoding 240 ORFs and 23 tRNA genes. Based on virion structure and genomic composition, Hena1 was classified as a new species of bacteriophage subfamily Vequintavirinae. The comprehensive analysis of Hena1 genome may provide further insights into evolution of bacteriophages infecting plant pathogenic bacteria.

scholarly journals Detection of Fire Blight disease in pear trees by hyperspectral data

2017 ◽  
Vol 51 (1) ◽  
pp. 1-10 ◽  
Author(s):  
Nikrooz Bagheri ◽  
Hosna Mohamadi-Monavar ◽  
Aslan Azizi ◽  
Abolghasem Ghasemi
Keyword(s):  
Fire Blight ◽  
Hyperspectral Data ◽  
Pear Trees ◽  
Blight Disease

scholarly journals Evaluation of Susceptibility of Different Pear Hybrid Populations to Fire Blight (Erwinia amylovora)

2011 ◽  
Vol 39 (1) ◽  
pp. 226 ◽  
Author(s):  
Yasemin EVRENOSOĞLU ◽  
Adalet MISIRLI ◽  
Hikmet SAYGILI ◽  
Emre BİLEN ◽  
Özlem BOZTEPE ◽  
...  
Keyword(s):  
Fire Blight ◽  
Erwinia Amylovora ◽  
Open Pollination ◽  
Resistance Level ◽  
Class A ◽  
Class B ◽  
Artificial Inoculations ◽  
Serious Disease ◽  
Blight Disease ◽  
Santa Maria

Fire blight disease caused by pathogenic bacterium Erwinia amylovora, is the serious disease of pear, and there is not a certain chemical management against this disease except antibiotic-type compounds such as streptomycin. It is very important to improve new fire blight resistant cultivars in case of integrated disease management. With this purpose, different crosses have been made between Pyrus communis varieties that have good fruit characteristics and resistant cultigens. Besides, self and open pollination treatments have been carried out in maternal plants. The disease resistance level of the hybrids obtained from these combinations was determined by artificial inoculations by Erwinia amylovora in greenhouse conditions. A total of 3284 hybrids were inoculated, and 2631 of them survived and were distributed to different susceptibility classes. 19.88% of the inoculated hybrids was killed by Erwinia amylovora. Total distribution of the hybrids to susceptibility classes was as 6.18% in class “A- slightly susceptible”, 3.11% in class “B- less susceptible”, 8.89% in class “C- mid-susceptible”, 20.28% in class “D- susceptible”, and 61.54% in class “E- very susceptible”. Majority of class “A- slightly susceptible” hybrids were obtained from ‘Magness’ x ‘Ankara’ combination. ‘Kieffer’ x ‘Santa Maria’, ‘Kieffer’ open pollination, ‘Magness’ x ‘Akça’, ‘Magness’ x ‘Kieffer’, ‘Magness’ x ‘Santa Maria’, ‘Mustafa Bey’ x ‘Moonglow’ treatments displayed good results with respect to “A- slightly susceptible” character. It is very important to evaluate these hybrid pear populations through different fruit and tree characteristics in the future.

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