In-orchard population dynamics of Erwinia amylovora on apple flower stigmas

Populations of the fire blight pathogen Erwinia amylovora Ea110 on apple flower stigmas were tracked over the course of apple bloom in field studies conducted between 2016 and 2019. In 18 of 23 experiments, flower stigmas inoculated on the 1st day of opening were found to harbor large (106-107 cells / flower) populations of E. amylovora when assessed three to five days post-inoculation. However, populations inoculated on stigmas of flowers that were already open for three days did not reach 106 cells / flower, and populations inoculated on stigmas of flowers that were already open for five days never exceeded 104 cells / flower. During this study, >10-fold increases in E. amylovora stigma populations in a 24-hr time period (termed population surges) were observed on 34.8%, 20.0%, and 4.0% of possible days on 1-day, 3-day, and 5-day open flowers, respectively. Population surges occurred on days with average temperatures as high as 24.5°C and as low as 6.1°C. Experiments incorporating more frequent sampling during days and overnight revealed that many population surges occurred between 10:00 PM and 2:00 AM. A Pearson’s correlation analysis of weather parameters occurring during surge events indicated that population surges were significantly associated with situations where overnight temperatures either increased or remained constant, where wind speed decreased, and where relative humidity increased. This study refines our knowledge of E. amylovora population dynamics and further indicates that E. amylovora is able to infect flowers during exposure to colder field temperatures than previously reported.

Antagonistic Activity of Bacillus spp. Against Fire Blight Disease In vitro and In planta

Fire blight, affecting more than one hundred and thirty species in the Rosaceae, is probably the most destructive disease affecting pear and apple cultivars in many countries. Currently, there are no effective synthetic compounds with systemic properties. Other major problem is the occurrence and spread of strains of Erwinia amylovora with resistance to streptomycin and copper. Taken into consideration the human and environmental health, the use of biocontrol agents either as an alternative or as a supplement within an integrated fire blight management strategy has attracted worldwide attention. In this study, E. amylovora solution of 107 CFU ml-1 was treated with bio-control agents, Bacillus subtilis str. QST 713, B. amyloliquefaciens str. MBI 600 and their mixture (at solution densities of 106, 107 and 108 CFU ml-1 for each one) on Petri dishes, containing King’s B medium and, compared with positive (streptomycin sulphate) and negative (sterile distilled water) controls. In vivo studies were performed on two-year-old apple cv. Gala seedlings grown in 45-cm-diameter pots containing a sterilized mix of soil–sand–peat under controlled greenhouse conditions (85% relative humidity, 25°C temperature and 16h of day light). The plants were irrigated as needed by drip-irrigation and each pot received a mineral solution (NPK: 20–20–20) at 2 g l-1 twice. When plant shoots reached a length of 30-35 cm, bio-control agents, individually and their mixture, were applied to the plants by a hand-sprayer. Obtaining the data, 108 CFU ml-1 of Bacillus spp. suspension mixture showed strongest in vitro antibacterial effect (26mm) among the tested treatments after positive control streptomycin (28.6mm). Parallel to in vitro findings, the mixture was most effective against the pathogen on cv. Gala (66.03%). Findings show that the use of mixture of beneficial microorganisms with individual antagonistic properties against the pathogen can be an effective strategy as a natural alternative to agrochemicals in the scope of good agriculture practices.

Draft Genome Sequences of Four Streptomycin-Sensitive Erwinia amylovora Strains Isolated from Commercial Apple Orchards in Ohio

Erwinia amylovora is the causative agent of fire blight, a devastating disease of apples and pears worldwide. Here, we report draft genome sequences of four streptomycin-sensitive strains of E. amylovora that were isolated from diseased apple trees in Ohio.

Application of the Maryblyt Model for the Infection of Fire Blight on Apple Trees at Chungju, Jecheon, and Eumsung during 2015-2020

To preventively control fire blight in apple trees and determine policies regarding field monitoring, the Maryblyt ver. 7.1 model (MARYBLYT) was evaluated in the cities of Chungju, Jecheon, and Eumseong in Korea from 2015 to 2020. The number of blossom infection alerts was the highest in 2020 and the lowest in 2017 and 2018. And the common feature of MARYBLYT blossom infection risks during the flowering period was that the time of BIR-High or BIR-Infection alerts was the same regardless of location. The flowering periods of the trees required to operate the model varied according to the year and geographic location. The model predicts the risk of “Infection” during the flowering periods, and recommends the appropriate times to control blossom infection. In 2020, when flower blight was severe, the difference between the expected date of blossom blight symptoms presented by MARYBLYT and the date of actual symptom detection was only 1-3 days, implying that MARYBLYT is highly accurate. As the model was originally developed based on data obtained from the eastern region of the United States, which has a climate similar to that of Korea, this model can be used in Korea. To improve field utilization, however, the entire flowering period of multiple apple varieties needs to be considered when the model is applied. MARYBLYT is believed to be a useful tool for determining when to control and monitor apple cultivation areas that suffer from serious fire blight problems.

Evaluation of Three Antimicrobial Peptides Mixtures to Control the Phytopathogen Responsible for Fire 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.

Refinement of Non-Antibiotic Spray Programs for Fire Blight Control in Organic Pome Fruit

Fire blight-susceptible, certified organic pome fruit is produced currently on 9,000 ha in the Pacific Northwest region of the United States with acreage continuing to expand in spite of a 2014 prohibition on antibiotics as allowable materials for infection suppression. Non-antibiotic practices for fire blight suppression mirror conventional management, but the full bloom to petal fall period when antibiotics are typically sprayed for fire blight control continues to receive research scrutiny owing to drawbacks and weaknesses of alternative materials. As solitary treatments, effective non-antibiotic materials (e.g., a yeast biocontrol, soluble coppers, and alum) raise the risk of a crop-value reducing, phytotoxic response termed fruit russeting. Conversely, materials with less russeting risk (e.g., Bacillus-based biorationals) are less effective for fire blight control. Spray programs using a combination of materials applied from mid-bloom to petal fall have the potential to provide high levels of protection with reduced russeting risk. In orchard trials, the effects of non-antibiotic spray programs on epiphytic population size of Erwinia. amylovora in flowers, yeast biocontrol population size, floral pH, infection suppression, and fruit russeting revealed strategies for sequencing sprays of non-antibiotic materials. The yeast biocontrol, Blossom Protect (Aureobasidium pullulans), sprayed at 70% bloom was an important contributor to fire blight suppression as was the soluble copper material, Previsto, when applied at full bloom. Choice of material for the petal fall spray timing was important to fruit russeting risk but apparently less important to overall infection incidence. Consequently, treatment programs of Blossom Protect at 70% bloom, a soluble copper at full bloom, and a Bacillus-based biorational at petal fall best balances the quality of infection suppression with risk of fruit russeting.