scholarly journals Effectiveness of Kasugamycin Against Erwinia amylovora and its Potential Use for Managing Fire Blight of Pear

Plant Disease ◽  
2011 ◽  
Vol 95 (4) ◽  
pp. 448-454 ◽  
Author(s):  
J. E. Adaskaveg ◽  
H. Förster ◽  
M. L. Wade

Comparative field studies on the management of fire blight were conducted in California on Asian and Bartlett pear using single-bactericide, mixture, and rotation treatments of selected compounds. Treatment efficacy was evaluated based on the natural occurrence of the disease or after inoculation with Erwinia amylovora. Kasugamycin at 100 mg/liter demonstrated similar or higher pre- and post-infection activity than the industry standards, streptomycin and oxytetracycline. Phytotoxicity caused by kasugamycin was observed only when five or six sequential weekly applications were done. In a six-spray rotation program including three bactericides (copper, kasugamycin, and oxytetracycline), with each being used twice, phytotoxicity was minor. Baseline sensitivity concentrations for kasugamycin were established for growth of 376 isolates of E. amylovora from California. Values for the lowest concentration where a reduction in growth on nutrient agar was observed ranged from 3.5 to 18.3 mg/liter, with a mean value of 8.7 mg/liter. Values for ≥95% inhibition of growth ranged from 6.9 to 46.7 mg/liter, with a mean value of 18.5 mg/liter. These inhibitory values for kasugamycin were higher than those for streptomycin or oxytetracycline. The in vitro activity of all three compounds was highly dependent on the agar medium used in the sensitivity assay. The activity of kasugamycin was also highly dependent on the pH of the medium and was significantly higher at pH 5.1 than pH 7.3. With the planned registration in the United States, kasugamycin represents the first new, highly effective bactericide for managing fire blight in over 40 years.

Plant Disease ◽  
2015 ◽  
Vol 99 (5) ◽  
pp. 659-666 ◽  
Author(s):  
Amanda Saville ◽  
Kim Graham ◽  
Niklaus J. Grünwald ◽  
Kevin Myers ◽  
William E. Fry ◽  
...  

Phytophthora infestans causes potato late blight, an important and costly disease of potato and tomato crops. Seven clonal lineages of P. infestans identified recently in the United States were tested for baseline sensitivity to six oomycete-targeted fungicides. A subset of the dominant lineages (n = 45) collected between 2004 and 2012 was tested in vitro on media amended with a range of concentrations of either azoxystrobin, cyazofamid, cymoxanil, fluopicolide, mandipropamid, or mefenoxam. Dose-response curves and values for the effective concentration at which 50% of growth was suppressed were calculated for each isolate. The US-8 and US-11 clonal lineages were insensitive to mefenoxam while the US-20, US-21, US-22, US-23, and US-24 clonal lineages were sensitive to mefenoxam. Insensitivity to azoxystrobin, cyazofamid, cymoxanil, fluopicolide, or mandipropamid was not detected within any lineage. Thus, current U.S. populations of P. infestans remained sensitive to mefenoxam during the displacement of the US-22 lineage by US-23 over the past 5 years.


Plant Disease ◽  
2019 ◽  
Vol 103 (2) ◽  
pp. 331-337 ◽  
Author(s):  
Wayne M. Jurick ◽  
Otilia Macarisin ◽  
Verneta L. Gaskins ◽  
Wojciech J. Janisiewicz ◽  
Kari A. Peter ◽  
...  

Penicillium spp. cause blue mold of stored pome fruit. These fungi reduce fruit quality and produce mycotoxins that are regulated for processed fruit products. Control of blue mold is achieved by fungicide application, and in 2015 Academy (active ingredients fludioxonil and difenoconazole) was released for use on pome fruit to manage postharvest blue mold. Baseline sensitivity for fludioxonil but not difenoconazole has been determined for P. expansum. To establish the distribution of sensitivity to difenoconazole before commercial use of Academy, 97 unexposed single-spore isolates from the United States and abroad were tested in vitro. Baseline EC50 values ranged from 0.038 to 0.827 µg/ml of difenoconazole with an average of 0.16 µg/ml. Complete inhibition of mycelial growth for all but three isolates occurred at 5 µg/ml of difenoconazole, whereas 10 µg/ml did not support growth for any of the isolates examined. Hence, 5 µg/ml of difenoconazole is recommended for phenotyping Penicillium spp. isolates with reduced sensitivity. Isolates with resistance to pyrimethanil and to both thiabendazole and pyrimethanil were observed among the isolates from the baseline collection. Academy applied at the labeled rate had both curative and protectant activities and controlled four representative Penicillium spp. from the baseline population. This information can be used to monitor future shifts in sensitivity to this new postharvest fungicide in Penicillium spp. populations.


Plant Disease ◽  
2017 ◽  
Vol 101 (5) ◽  
pp. 738-743
Author(s):  
Achour Amiri ◽  
Aaron W. Hawkins ◽  
Katie A. Mulvaney

Very little is known about the biology, epidemiology, and best practices to manage Lambertella rot, a newly identified postharvest disease caused by Lambertella corni-maris on apples in the United States. In this study, we investigated the prevalence of L. corni-maris in 92 grower lots throughout Washington State in 2016, evaluated the effect of nutrient availability on L. corni-maris growth, and L. corni-maris sensitivity to pH, osmotic stress, and fungicides in vitro. We assessed pathogen aggressiveness on major apple cultivars and the efficacy of pre- and postharvest fungicides to control L. corni-maris on detached fruit. L. corni-maris was widespread and was found in 40% of the growers lots surveyed at frequencies ranging from 2 to 40% of the total decay. The fungus grew faster on acidic media such as apple juice agar and V8 agar media and was able to grow equally at pH values ranging from 3 to 7. L. corni-maris isolates showed relatively low sensitivity to osmotic stress and grew evenly at 1M KCl. All nine apple cultivars tested were susceptible to L. corni-maris, but the disease severity was significantly higher on some cultivars such as Honeycrisp, Piñata, and Gala. The baseline sensitivity of 100 L. corni-maris isolates was determined based on the effective concentration necessary to inhibit 50% mycelial growth (EC50) for fludioxonil, boscalid, pyraclostrobin + SHAM, pyrimethanil, and thiabendazole. The respective mean EC50 values were 0.07, 0.84, 1.32, 2.45, and 3.68 µg/ml. Fludioxonil and pyrimethanil applied at label rates were the only fungicides able to control L. corni-maris effectively on detached apple fruit. This study is a first step toward understanding the eco-physiological requirements for L. corni-maris to survive and cause decay on apple, assessing potential shifts in fungicide sensitivity, and enhancing Lambertella rot management. We suggest “yellow rot” as the new common name for Lambertella rot.


Author(s):  
Kubilay Kurtulus Bastas

Erwinia amylovora, the causative agent of fire blight disease, threatens a lot of species of the Rosaceae family. Antibiotics and copper compounds in chemical applications are most frequently are applied, but these can be phytotoxic and cause resistant strains of the pathogen. In our experiments, 20 herbal materials were tested for their antimicrobial effectiveness against the fire blight pathogen in vitro and in planta. The air-dried plants ground into fine powder and extraction was performed at room temperature by maceration with 80% (v/v) methanol/distilled water. The minimum inhibitory concentration values were determined by using disc diffusion method and streptomycin was used as control in all experiments. Antimicrobial activity was evaluated by measuring the inhibition zones in reference to the pathogen. Among the tested plants, Szygium aromaticum, Thymus vulgaris and Rhus cararia showed a good antibacterial activity and they inhibited the growth of E. amylovora with inhibition zone diameter ranging from 21 to 27 mm at 20% (w/v) in absolute methanol compared to streptomycin (31 mm) in vitro conditions. In vivo tests were performed by using highly virulent E. amylovora isolate (Eak24b, 91%) grown on TSA medium and inoculation on young shoots of 3-year-old Gala variety of apple and Santa Maria variety of pear seedlings at 107 CFU ml-1 density of the pathogen. Disease severity (%) was assessed by by proportion of blighted shoot length to the whole shoot length and also efficacy of the extracts was determined by using Abbott formula. The highest efficacy was obtained by S. aromaticum and T. vulgaris extracts of reducing shoot blight of cv. Gala and cv. Santa Maria by 67.81% - 64-12% and 51.50% - 51.04% ratios, respectively. Obtaining results showed that some medicinal and aromatic plant extracts might be used against fire blight disease as potential new generation chemicals on pome fruits within integrated and organic control programs.


Plant Disease ◽  
2016 ◽  
Vol 100 (4) ◽  
pp. 802-809 ◽  
Author(s):  
K. A. Tancos ◽  
S. Villani ◽  
S. Kuehne ◽  
E. Borejsza-Wysocka ◽  
D. Breth ◽  
...  

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.


Plant Disease ◽  
2011 ◽  
Vol 95 (2) ◽  
pp. 189-194 ◽  
Author(s):  
C. A. Bradley ◽  
D. K. Pedersen

Cercospora zeae-maydis, the causal agent of gray leaf spot on corn (Zea mays), can cause severe yield loss in the United States. Quinone outside inhibitor (QoI) fungicides are effective tools that can be used to manage gray leaf spot, and their use has increased in corn production in the United States. In total, 61 C. zeae-maydis isolates collected from fields in which QoI fungicides had never been applied were tested in vitro using azoxystrobin-, pyraclostrobin-, or trifloxystrobin-amended medium to determine the effective fungicide concentration at which 50% of the conidial germination was inhibited (EC50). The effect of salicylhydroxamic acid (SHAM) also was evaluated for seven isolates to determine whether C. zeae-maydis is capable of using alternative respiration in azoxystrobin-amended medium. All seven C. zeae-maydis isolates tested had significantly greater (P < 0.02) EC50 values when SHAM was not included in medium amended with azoxystrobin, indicating that C. zeae-maydis has the potential to utilize alternative respiration to overcome QoI fungicide inhibition in vitro. Baseline EC50 values of azoxystrobin ranged from 0.003 to 0.031 μg/ml, with mean and median values of 0.018 and 0.019 μg/ml, respectively. Baseline EC50 values of pyraclostrobin ranged from 0.0003 to 0.0025 μg/ml, with mean and median values of 0.0010 and 0.0010 μg/ml, respectively. Baseline EC50 values of trifloxystrobin ranged from 0.0004 to 0.0034 μg/ml, with mean and median values of 0.0023 and 0.0024 μg/ml, respectively. These baseline sensitivity values will be used in a fungicide resistance monitoring program to determine whether shifts in sensitivity to QoI fungicides are occurring in C. zeae-maydis populations.


2011 ◽  
Vol 101 (2) ◽  
pp. 192-204 ◽  
Author(s):  
Gayle C. McGhee ◽  
George W. Sundin

The emergence and spread of streptomycin-resistant strains of Erwinia amylovora in Michigan has necessitated the evaluation of new compounds effective for fire blight control. The aminoglycoside antibiotic kasugamycin (Ks) targets the bacterial ribosome and is particularly active against E. amylovora. The efficacy of Ks formulated as Kasumin 2L for control of fire blight was evaluated in six experiments conducted over four field seasons in our experimental orchards in East Lansing, MI. Blossom blight control was statistically equivalent to the industry standard streptomycin in all experiments. E. amylovora populations remained constant on apple flower stigmas pretreated with Kasumin and were ≈100-fold lower than on stigmas treated with water. Kasumin applied to apple trees in the field also resulted in a 100-fold reduced total culturable bacterial population compared with trees treated with water. We performed a prospective analysis of the potential for kasugamycin resistance (KsR) development in E. amylovora which focused on spontaneous resistance development and acquisition of a transferrable KsR gene. In replicated lab experiments, the development of spontaneous resistance in E. amylovora to Ks at 250 or 500 ppm was not observed when cells were directly plated on medium containing high concentrations of the antibiotic. However, exposure to increasing concentrations of Ks in media (initial concentration 25 μg ml–1) resulted in the selection of Ks resistance (at 150 μg ml–1) in the E. amylovora strains Ea110, Ea273, and Ea1189. Analysis of mutants indicated that they harbored mutations in the kasugamycin target ksgA gene and that all mutants were impacted in relative fitness observable through a reduced growth rate in vitro and decreased virulence in immature pear fruit. The possible occurrence of a reservoir of KsR genes in orchard environments was also examined. Culturable gram-negative bacteria were surveyed from six experimental apple orchards that had received at least one Kasumin application. In total, 401 KsR isolates (42 different species) were recovered from apple flowers and leaves and orchard soil samples. Although we have not established the presence of a transferrable KsR gene in orchard bacteria, the frequency, number of species, and presence of KsR enterobacterial species in orchard samples suggests the possible role of nontarget bacteria in the future transfer of a KsR gene to E. amylovora. Our data confirm the importance of kasugamycin as an alternate antibiotic for fire blight management and lay the groundwork for the development and incorporation of resistance management strategies.


2011 ◽  
Vol 101 (10) ◽  
pp. 1234-1241 ◽  
Author(s):  
P. L. Pusey ◽  
V. O. Stockwell ◽  
C. L. Reardon ◽  
T. H. M. Smits ◽  
B. Duffy

Pantoea agglomerans E325, the active ingredient in a commercial product for fire blight control, was previously shown in vitro to produce a unique alkaline- and phosphate-sensitive antibiotic specific to Erwinia amylovora. Antibiosis was evaluated as a mode of antagonism on flower stigmas using two antibiosis-deficient mutants. On King's medium B, mutants E325ad1 and E325ad2 have stable smooth-butyrous or hypermucoid colony morphologies, respectively, and the parental strain E325 exhibits phenotypic plasticity with predominantly hypermucoid colonies accompanied by slower-growing, smooth-butyrous colonies. Mutants were tested against E. amylovora on stigmas of detached flowers of crab apple (Malus mandshurica) in growth chambers and apple (Malus domestica) in the orchard. Epiphytic fitness of the antibiosis-negative mutants was similar or greater than the parental strain as determined by relative area under the population curve (RAUPC). In laboratory and orchard trials, both mutants had significantly lower inhibitory activity against the pathogen (i.e., less reduction of E. amylovora RAUPC) compared with the parental strain. E325 and the mutants caused similar decreases in pH in a broth medium, indicating that acidification, which was previously reported as a possible mechanism of pathogen inhibition on stigmas, is not directly related to antibiosis. In this study we provide the first evidence for E325 antibiosis involved in E. amylovora growth suppression on apple flower stigmas.


Plant Disease ◽  
2008 ◽  
Vol 92 (2) ◽  
pp. 295-300 ◽  
Author(s):  
K. A. Wise ◽  
C. A. Bradley ◽  
J. S. Pasche ◽  
N. C. Gudmestad ◽  
F. M. Dugan ◽  
...  

Ascochyta rabiei, causal agent of Ascochyta blight on chickpea (Cicer arietinum), can cause severe yield loss in the United States. Growers rely on applications of fungicides with site-specific modes of action such as the quinone outside inhibiting (QoI) fungicides azoxystrobin and pyraclostrobin, and the carboximide fungicide boscalid, to manage disease. In all, 51 isolates collected prior to QoI fungicide registration and 71 isolates collected prior to boscalid registration in the United States were tested in an in vitro assay to determine the effective fungicide concentration at which 50% of conidial germination was inhibited (EC50) for each isolate–fungicide combination. The effect of salicylhydroxamic acid (SHAM) on conidia of A. rabiei in the presence and absence of azoxystrobin also was assessed to determine whether the fungus is capable of using alternative respiration. Five of nine A. rabiei isolates tested had significantly higher (P ≤ 0.05) EC50 values when SHAM was not included in media amended with azoxystrobin, indicating that A. rabiei has the potential to use alternative respiration to overcome fungicide toxicity in vitro. EC50 values of azoxystrobin and pyraclostrobin ranged from 0.0182 to 0.0338 μg/ml and from 0.0012 to 0.0033 μg/ml, with mean values of 0.0272 and 0.0023 μg/ml, respectively. EC50 values of boscalid ranged from 0.0177 to 0.4960 μg/ml, with a mean of 0.1903 μg/ml. Establishment of these baselines is the first step in developing a monitoring program to determine whether shifts in sensitivity to these fungicides are occurring in the A. rabiei pathogen population.


Pathogens ◽  
2020 ◽  
Vol 9 (1) ◽  
pp. 41 ◽  
Author(s):  
Neelendra K. Joshi ◽  
Henry K. Ngugi ◽  
David J. Biddinger

Fire blight, which is caused by the bacteria Erwinia amylovora, remains one of the most important diseases limiting the productivity of apple and pear orchards in the United States. In commercial orchards, in-season fire blight management relies exclusively on the use of antibiotic treatments (such as streptomycin and oxytetracycline) and on bacterial biocontrol agents whose efficacy is limited. We hypothesize that the efficacy of the biocontrol agents can be greatly enhanced through targeted delivery to flowers, which serve as initial infection courts, using the Japanese orchard bee, Osmia cornifrons. Many factors, such as the synchrony of life cycle with plant phenology and specificity to pomaceous plants, suggest that O. cornifrons could be an excellent vector of the biocontrol products during bloom in pome tree fruits. However, deployment of this pollinator species to deliver biocontrol agents for fire blight control has not been attempted previously due to the lack of an efficient system to pack the bodies of the bees exiting nesting tubes with the biocontrol products. In this study, we design and test a dispenser system to facilitate the use of O. conifrons as a vector for commercially available biocontrol products for fire blight control. The effectiveness of O. conifrons to deliver biocontrol agents to flowers, and to effect secondary dissemination from treated to untreated flowers is also evaluated in greenhouse experiments. We found that the O. conifrons bees were able to use the nest dispenser designed for the delivery of biological control products, and are effective in vectoring and delivering the Bacillus subtilis-based biological control product (Serenade®) to apple blossoms. We also found that the O. cornifrons were effective in secondary inoculation of this biological control product to newly-opened flowers. These findings suggest the potential use of commercially available O. conifrons and other orchard bees in targeted delivery of biological control products for fire blight, and possibly other diseases, in different fruit crops.


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