The Effect of Erwinia amylovora Infection in Apple Saplings and Fruit on the Behavior of Delia platura (Diptera: Anthomyiidae)

2020 ◽  
Author(s):  
Matthew Boucher ◽  
Rowan Collins ◽  
Stephen Hesler ◽  
Kerik Cox ◽  
Greg Loeb
Keyword(s):  
Erwinia Amylovora ◽  
Bacterial Pathogen ◽  
Fruit Flies ◽  
Pome Fruit ◽  
Enhanced Transmission ◽  
Diseased Fruit ◽  
Fruit Odor ◽  
Shoot Blight ◽  
Insect Interactions ◽  
Delia Platura

Abstract The Vector Manipulation Hypothesis (VMH) posits that phytopathogens develop strategies to enhance dissemination by mediating behavior change in insect vectors. The VMH is poorly studied in phytopathogenic bacteria, especially in systems with numerous, occasional vectors. Erwinia amylovora is a bacterial pathogen of pome fruit that produces a bacterial ooze and is mechanically vectored by insects after they feed on ooze. The blossom blight phase of the disease exhibits manipulation of honeybees, leading to enhanced transmission, but whether the same occurs during the shoot blight phase of the disease is unknown. The goal of this study was to evaluate the effect of E. amylovora on the behavior of Delia platura, a fly with a worldwide endemic presence that may transmit E. amylovora. We show that D. platura prefer infected, oozing fruit to uninfected fruit in choice tests and that preference subsides when bacterial ooze is removed from the infected fruit. Flies did not exhibit a preference between infected saplings and uninfected saplings. The volatiles of infected fruit did not attract D. platura, indicating that diseased fruit odor is not responsible for the observed preference for infected fruit. Flies did not differentiate between sapling odors until infected trees had died, at which point they preferred uninfected tree odors. This study supports previous hypotheses suggesting that E. amylovora takes advantage of existing plant–insect interactions, though it is not fully understood how significantly behavioral changes affect transmission. Additional pathosystems with occasional, nonspecific vectors should be studied to further understanding of the VMH.

scholarly journals Interactions Between Delia platura and Erwinia amylovora Associated with Insect Mediated Transmission of Shoot Blight

2020 ◽  
pp. PHYTOFR-08-20-0 ◽  
Author(s):  
Matthew Boucher ◽  
Rowan Collins ◽  
Kayli Harling ◽  
Gabrielle Brind’Amour ◽  
Kerik Cox ◽  
...  
Keyword(s):  
Open Access ◽  
Plant Defense ◽  
Erwinia Amylovora ◽  
Bacterial Pathogen ◽  
Biofilm Production ◽  
Daily Rate ◽  
Shoot Blight ◽  
First Time ◽  
Delia Platura ◽  
Open Access Article

Erwinia amylovora is a bacterial pathogen of rosaceous plants that can be devastating to commercial apple and pear production worldwide. Exopolysaccharide (EPS) production is essential for pathogenicity, aiding in biofilm production, and plant defense protection. EPS also plays an epidemiological role in the form of bacterial ooze, which is generated when E. amylovora builds to such high levels within the parenchyma that it ruptures the plant epidermis and a mixture of bacteria encased in EPS exudes from the injury. Insects such as Delia platura feed on the ooze and become potential vectors in the process. The goal of this study was to investigate interactions between D. platura and E. amylovora to better understand how insects facilitate shoot blight. We demonstrate for the first time that D. platura can successfully transmit pathogen cells that initiate new infections in mechanically damaged apple shoots and that EPS aids in adherence of E. amylovora to the insect surface. We show that flies can carry the bacteria externally for at least 5 days and that they shed a constant daily rate. We also show that E. amylovora strains differing in virulence do not behave differently when associated with D. platura and that consumption of E. amylovora has no effect on insect survival. Our data demonstrate that flies can be efficient facilitators of new shoot blight infections, but the field conditions that make this type of transmission possible require further investigation. [Formula: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

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.

Phylogenetic analysis of PR genes in some pome fruit species with the emphasis on transcriptional analysis and ROS response under Erwinia amylovora inoculation in apple

Genetica ◽  
2015 ◽  
Vol 144 (1) ◽  
pp. 9-22 ◽  
Author(s):  
Maryam Hassani ◽  
Seyed Alireza Salami ◽  
Jaber Nasiri ◽  
Hamid Abdollahi ◽  
Zahra Ghahremani
Keyword(s):  
Phylogenetic Analysis ◽  
Erwinia Amylovora ◽  
Transcriptional Analysis ◽  
Pome Fruit ◽  
Pr Genes ◽  
Fruit Species

scholarly journals Blackberry and raspberry are alternative resistance sources to fire blight

2017 ◽  
Vol 97 (1) ◽  
pp. 12-16 ◽  
Author(s):  
Ozer Calis ◽  
Cetin Cekic ◽  
Serhat Kara ◽  
Demet Celik Ertekin
Keyword(s):  
Bacterial Growth ◽  
Fire Blight ◽  
Erwinia Amylovora ◽  
Bacterial Pathogen ◽  
Post Inoculation ◽  
Resistance Sources ◽  
Pathogenicity Tests ◽  
Susceptibility And Resistance ◽  
Resistant Genotypes ◽  
Moderate Susceptibility

Erwinia amylovora causes fire blight mainly on pear, apple and quince trees. This bacterial pathogen also infects other Rosaceous plants, such as blackberry and raspberry. A race structure was established between an isolate of E. amylovora and berries using 40 wild blackberry and 7 wild raspberry genotypes. In pathogenicity tests, wild blackberry and raspberry genotypes had three phenotypic reactions: enhanced susceptibility, moderate susceptibility and resistance. We noted a higher bacterial growth of over 300 × 109 cfu mL-1 in plants with enhanced susceptibility, with resistant genotypes showing a bacterial growth of around 150 × 109 cfu mL-1. These results are also associated with symptoms observed at 29 days post-inoculation. This resistance is being evaluated to control fire blight.

scholarly journals CRISPR genotyping as complementary tool for epidemiological surveillance of Erwinia amylovora outbreaks

PLoS ONE ◽  
2021 ◽  
Vol 16 (4) ◽  
pp. e0250280
Author(s):  
Rafael J. Mendes ◽  
João Pedro Luz ◽  
Conceição Santos ◽  
Fernando Tavares
Keyword(s):  
Fire Blight ◽  
Erwinia Amylovora ◽  
Blot Hybridization ◽  
Fruit Trees ◽  
Epidemiological Surveillance ◽  
Economic Losses ◽  
Pome Fruit ◽  
Dot Blot ◽  
Virulence Markers ◽  
Clonal Population Structure

Fire blight is a destructive plant disease caused by Erwinia amylovora affecting pome fruit trees, and responsible for large yield declines, long phytosanitary confinements, and high economic losses. In Portugal, the first major fire blight outbreaks occurred in 2010 and 2011, and although later considered eradicated, the emergence of other outbreaks in recent years stressed the need to characterize the E. amylovora populations associated with these outbreaks. In this regard, CRISPR genotyping, assessment of three virulence markers, and semi-quantitative virulence bioassays, were carried out to determine the genotype, and assess the virulence of thirty-six E. amylovora isolates associated with outbreaks occurring between 2010 and 2017 and affecting apple and pear orchards located in the country central-west, known as the main producing region of pome fruits in Portugal. The data gathered reveal that 35 E. amylovora isolates belong to one of the widely-distributed CRISPR genotypes (5-24-38 / D-a-α) regardless the host species, year and region. Ea 680 was the single isolate revealing a new CRISPR genotype due to a novel CR2 spacer located closer to the leader sequence and therefore thought to be recently acquired. Regarding pathogenicity, although dot-blot hybridization assays showed the presence of key virulence factors, namely hrpL (T3SS), hrpN (T3E) and amsG from the amylovoran biosynthesis operon in all E. amylovora isolates studied, pathogenicity bioassays on immature pear slices allowed to distinguish four virulence levels, with most of the isolates revealing an intermediate to severe virulence phenotype. Regardless the clonal population structure of the E. amylovora associated to the outbreaks occurring in Portugal between 2010 and 2017, the different virulence phenotypes, suggests that E. amylovora may have been introduced at different instances into the country. This is the first study regarding E. amylovora in Portugal, and it discloses a novel CRISPR genotype for this bacterium.

scholarly journals Erwinia amylovora Enfeksiyonu Sonrası Elma, Armut ve Ayva Çeşitlerinde Konukçu Protein Miktarlarının Belirlenmesi

2014 ◽  
Vol 3 (3) ◽  
pp. 154
Author(s):  
Şerife Çetin ◽  
Kubilay Kurtuluş Baştaş
Keyword(s):  
Protein Content ◽  
Total Protein ◽  
Erwinia Amylovora ◽  
Bacterial Pathogen ◽  
Molecular Weights ◽  
Molecular Tests ◽  
Sds Page ◽  
Apple Cultivars ◽  
Blight Disease ◽  
Santa Maria

Fire blight disease caused by Erwinia amylovora is a destructive bacterial pathogen mainly on pears, apples and quinces from Rosaceae family. In this study, it was aimed determination of total protein amounts in different apple cultivars (Braeburn, Fuji, Gala and Golden), pear cultivars (Santa Maria and Williams) and quince cultivars (Eşme and Ekmek) in the infections of two virulent E. amylovora strains (Ea234-1 and Ea240-3) according as the time. It was taken leaf samples after leaf inoculation with E. amylovora (108 CFU ml-1) at 24th, 36th and 72nd hours. For verification of the infections, re-isolations were made from bacteria inoculated plants and the agent was identified as E. amylovora by biochemical, physiological and molecular tests. In determining the amounts of total protein and in the SDS-PAGE analyses were used Bradford and Laemmli methods, respectively, and absorbance values of protein extracts derived from the leaf samples taken, were obtained at 595 nm wavelength. According to the findings obtained; after infection of E. amylovora in the apple varieties comparing to controls, total protein concentrations at 24th hours increased and a decrease in the amount of 36th to 72nd hours and Braeburn has the highest protein content was determined. In the pear varieties, while total protein concentrations at 24th and 36th hours increased, a decrease in the amount of 72nd hour, and Santa Maria variety has the highest protein content was detected. In the quince varieties, total protein concentrations at 72th hour increased and Eşme variety has the highest protein content was identified. As a result of SDS-PAGE analysis, protein fractions which have different molecular weights were obtained. The protein bands were defined approximately 55-70 kDa and 35-55 kDa molecule weight on apple and quince varieties, respectively and also approx. 55-70 kDa in pear varieties.

scholarly journals Activation of metabolic and stress responses during subtoxic expression of the type I toxin hok in Erwinia amylovora

2021 ◽  
Author(s):  
Jingyu Peng ◽  
Lindsay R. Triplett ◽  
George Sundin
Keyword(s):  
Cell Death ◽  
Stress Responses ◽  
Erwinia Amylovora ◽  
Physiological State ◽  
Fruit Trees ◽  
Toxin Gene ◽  
Type I ◽  
Pome Fruit ◽  
Membrane Rupture ◽  
High Level

Abstract Background: Toxin-antitoxin (TA) systems, abundant in prokaryotes, are composed of a toxin gene and its cognate antitoxin. Several toxins are implied to affect the physiological state and stress tolerance of bacteria in a population. We previously identified a chromosomally encoded hok-sok type I TA system in Erwinia amylovora, the causative agent of fire blight disease on pome fruit trees. A high-level induction of the hok gene was lethal to E. amylovora cells through unknown mechanisms. The molecular targets or regulatory roles of Hok were unknown.Results: Here, we examined the physiological and transcriptomic changes of Erwinia amylovora cells expressing hok at subtoxic levels that were confirmed to confer no cell death, and at toxic levels that resulted in killing of cells. In both conditions, hok caused membrane rupture and collapse of the proton motive force in a subpopulation of E. amylovora cells. We demonstrated that induction of hok resulted in upregulation of ATP biosynthesis genes, and caused leakage of ATP from cells only at toxic levels. We showed that overexpression of the phage shock protein gene pspA largely reversed the cell death phenotype caused by high levels of hok induction. We also showed that induction of hok at a subtoxic level rendered a greater proportion of stationary phase E. amylovora cells tolerant to the antibiotic streptomycin. Conclusions: We characterized the molecular mechanism of toxicity by high-level of hok induction and demonstrated that low-level expression of hok primes the stress responses of E. amylovora against further membrane and antibiotic stressors.

scholarly journals Prevalence of Streptomycin-Resistant Erwinia amylovora in New York Apple Orchards

Plant Disease ◽  
2016 ◽  
Vol 100 (4) ◽  
pp. 802-809 ◽  
Author(s):  
K. A. Tancos ◽  
S. Villani ◽  
S. Kuehne ◽  
E. Borejsza-Wysocka ◽  
D. Breth ◽  
...  
Keyword(s):  
United States ◽  
New York ◽  
Fire Blight ◽  
Erwinia Amylovora ◽  
The United States ◽  
Streptomycin Resistance ◽  
Apple Orchards ◽  
Wayne County ◽  
The Past ◽  
Shoot Blight

Resistance to streptomycin in Erwinia amylovora was first observed in the United States in the 1970s but was not found in New York until 2002, when streptomycin-resistant (SmR) E. amylovora was isolated from orchards in Wayne County. From 2011 to 2014, in total, 591 fire blight samples representing shoot blight, blossom blight, and rootstock blight were collected from 80 apple orchards in New York. From these samples, 1,280 isolates of E. amylovora were obtained and assessed for streptomycin resistance. In all, 34 SmR E. amylovora isolates were obtained from 19 individual commercial orchards. The majority of the resistant isolates were collected from orchards in Wayne County, and the remaining were from other counties in western New York. Of the 34 resistant isolates, 32 contained the streptomycin resistance gene pair strA/strB in the transposon Tn5393 on the nonconjugative plasmid pEA29. This determinant of streptomycin resistance has only been found in SmR E. amylovora isolates from Michigan and the SmR E. amylovora isolates discovered in Wayne County, NY in 2002. Currently, our data indicate that SmR E. amylovora is restricted to counties in western New York and is concentrated in the county with the original outbreak. Because the resistance is primarily present on the nonconjugative plasmid, it is possible that SmR has been present in Wayne County since the introduction in 2002, and has spread within and out of Wayne County to additional commercial growers over the past decade. However, research is still needed to provide in-depth understanding of the origin and spread of the newly discovered SmR E. amylovora to reduce the spread of streptomycin resistance into other apple-growing regions, and address the sustainability of streptomycin use for fire blight management in New York.

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