Determining the Main Infection Courts in Sweet Cherry Trees of the Canker Pathogens Calosphaeria pulchella, Cytospora sorbicola, and Eutypa lata.

The major fungal canker pathogens causing branch dieback of sweet cherry trees in California include Calosphaeria pulchella, Cytospora sorbicola, and Eutypa lata. These pathogens have long been known to infect cherry trees mainly through pruning wounds. However, recent field observations revealed numerous shoots and fruiting spurs exhibiting dieback symptoms with no apparent pruning wounds or mechanical injuries. Accordingly, this study was conducted to assess the incidence of the three pathogens in symptomatic terminal shoots and dying fruiting spurs, in addition to the wood below pruning wounds in branches. Surveys were conducted in five sweet cherry orchards across three counties in California. We also investigated the possibility that leaf scars, bud scars, and wounds resulting from fruit picking could serve as infection courts for Cal. pulchella, Cyt. sorbicola, and E. lata by means of artificial inoculations in the field. Orchard surveys revealed that Cal. pulchella had the highest pathogen incidence below pruning wounds in branch samples, followed by Cyt. sorbicola and E. lata. Among terminal shoots with dieback symptoms and dying fruiting spurs, Cyt. sorbicola was the most prevalent, followed by Cal. pulchella. Results from field inoculations indicated that fruit-picking wounds could serve as important infection courts for Cal. pulchella, Cyt. sorbicola, and E. lata, with average pathogen recovery of 41.5, 63, and 36.2%, respectively. Results also indicated that leaf and bud scars could serve as an entry site for Cyt. sorbicola, although recovery was relatively low. The present study is the first to identify harvest-induced wounds on fruiting spurs of sweet cherry as an important infection court of Cal. pulchella, Cyt. sorbicola, and E. lata.

Factors Affecting the Infection of Sweet Cherry (Prunus avium) Fruit by Podosphaera cerasi.

Powdery mildew caused by Podosphaera cerasi is the most important fungal disease of sweet cherries in the Pacific Northwest of the United States. In this study, several factors related to disease epidemiology were evaluated. The experiments were conducted to investigate flower susceptibility to P. cerasi infection by in planta and in vitro inoculation. The susceptibility of fruit at various developmental stages was investigated using defined concentrations of P. cerasi conidia. Furthermore, the threshold of conidial concentration required for fruit infection was determined. The pathogen activity during full bloom was limited and not related to fruit disease incidence and severity at harvest. Foliar infections always preceded fruit infections by an average of 42 days during the 3 years of the study. The onset of fruit infection followed, on average, 66 days after full bloom and appeared simultaneously on all susceptible cherry cultivars in the research orchard. Disease symptoms were only observed on fruit in Biologische Bundesanstalt, Bundessortenamt, and Chemical Industry scale 8 (maturity) in all cultivars examined. During this stage, a concentration of 500 conidia/ml was sufficient to cause fruit infection at harvest. Interaction between the inoculation dates and conidial concentration revealed a dependency of disease development on the host stage at the time of inoculation; the younger the fruit, the more conidia are needed to cause disease at harvest. Molecular studies showed a rapid increase in conidia viability at the transition from asymptomatic to the symptomatic disease of fruit. No evidence of ontogenic resistance of fruit to powdery mildew infection was observed.

Integration of metabolomics and existing omics data reveals new insights into phytoplasma-induced metabolic reprogramming in host plants.

Phytoplasmas are cell wall-less bacteria that induce abnormal plant growth and various diseases, causing severe economic loss. Phytoplasmas are highly dependent on nutrients imported from host cells because they have lost many genes involved in essential metabolic pathways during reductive evolution. However, metabolic crosstalk between phytoplasmas and host plants and the mechanisms of phytoplasma nutrient acquisition remain poorly understood. In this study, using metabolomics approach, sweet cherry virescence (SCV) phytoplasma-induced metabolite alterations in sweet cherry trees were investigated. A total of 676 metabolites were identified in SCV phytoplasma-infected and mock inoculated leaves, of which 187 metabolites were differentially expressed, with an overwhelming majority belonging to carbohydrates, fatty acids/lipids, amino acids, and flavonoids. Available omics data of interactions between plant and phytoplasma were also deciphered and integrated into the present study. The results demonstrated that phytoplasma infection promoted glycolysis and pentose phosphate pathway activities, which provide energy and nutrients, and facilitate biosynthesis of necessary low-molecular metabolites. Our findings indicated that phytoplasma can induce reprograming of plant metabolism to obtain nutrients for its own replication and infection. The findings from this study provide new insight into interactions of host plants and phytoplasmas from a nutrient acquisition perspective.

Luteolin-induced activation of the phenylpropanoid metabolic pathway contributes to quality maintenance and disease resistance of sweet cherry.

Redox imbalance and fungal infection are major causes for quality deterioration and postharvest decay of fruit. Therefore, it is crucial to activate intrinsic antioxidative capacity and disease responses for fruit quality maintenance. Although plant-derived flavonoids have been reported for health-promoting benefits, their roles in the maintenance of fruit quality remains largely unexplored. Here, we exogenously applied luteolin, a flavonoid substance, and further examined its efficacy in maintaining fruit quality and inhibiting fungal diseases in sweet cherry. The results showed that 100 or 200 mg/L luteolin maintained better organoleptic quality and decreased disease incidence during storage. Biochemical assays revealed that luteolin activated the phenylpropanoid metabolic pathway and improved antioxidative capacity, thereby elevating total anthocyanin and flavonoid contents. Notably, luteolin inhibited mycelial growth of fungal pathogens and reduced patulin yield by Penicillium expansum. Collectively, these results suggest that luteolin is a promising alternative for maintaining better fruit quality and ameliorating disease resistance.

Antifungal activity of an allicin derivative against Penicillium expansum via induction of oxidative stress.

The application of natural preservatives has become an attractive method for controlling postharvest decay of fruits and vegetables. Allicin, the main active ingredient of allium plants, has broad-spectrum antifungal activity. However, the unstable properties of allicin limit its wide application. In this study, 1-[(R)-ethylsulfinyl]sulfanylethane, a structurally stable derivative of allicin, was used to explore its antifungal activity and potential mechanism on the expansion of Penicillium expansum. We demonstrated the antifungal activity of 1-[(R)-ethylsulfinyl]sulfanylethane through in vitro antifungal experiments. At the concentration is 2 mg/L, 1-[((R)-ethylsulfinyl]sulfanyl]ethane can completely inhibit spore germination and mycelial growth, whereas the concentration of allicin needs to reach 16 mg/L. Fungal Biochemical assay indicated that decrease of mitochondrial membrane potential, overgeneration of reactive oxygen species, decrease of adenosine triphosphate and glutathione content, increase of superoxide dismutase, catalase, and malondialdehyde content. The results revealed that 1-[(R)-ethylsulfinyl]sulfanylethane induced mitochondrial dysfunction and oxidative stress in P. expansum. Due to its excellent antifungal activity, 1-[((R)-ethylsulfinyl]sulfanyl]ethane might be developed as a substitute for fungicides against P. expansum in postharvest fruit storage.

Selection and application of antifungal VOCs-producing yeasts as biocontrol agents of grey mould in fruits.

Rotting caused by grey mould (Botrytis cinerea) is a concerning disease for numerous crops both pre- and postharvest stages. Application of antagonistic yeasts is a promising strategy for controlling grey mould incidence which could mitigate undesirable consequences of using synthetic fungicides. In this work, a screening for detection of yeasts isolated from figs producers of antifungal volatile organic compounds (VOCs) were performed by confrontation in double dishes systems. Eleven out of 34 yeasts confronted reduced B. cinerea growth parameter in vitro. This reduction was correlated (p ≤ 0.050) with the production of 10 volatile compounds: two acids (acetic acid and octanoic acid), 7 esters (Ethyl propionate, n-Propyl acetate, Isobutyl acetate, 2-methylbutyl acetate, furfuryl acetate, phenylmethyl acetate, 2-phenylethyl acetate) and one ketone (Heptan-2-one). In bases on in vitro assay, Hanseniaspora uvarum 793 was applied to in vivo assays with strawberries and cherries. The reduction of incidence of B. cinerea in strawberries at 7 °C and 25 °C was 54.9 and 72.1% after 6 and 3 days, respectively. The reduction of incidence of B. cinerea in cherries at 7 °C and 25 °C was 48.9 and 45.6% after 5 and 4 days, respectively. These results showed that VOCs produced by Hanseniaspora uvarum 793 are effective in the control of incidence of Botrytis cinerea in fruits, being a potential alternative to chemical fungicide.

Salmonella inactivation and cross-contamination on cherry and grape tomatoes under simulated wash conditions.

Washing in chlorinated water is widely practiced for commercial fresh produce processing. While known as an effective tool for mitigating food safety risks, chlorine washing could also represent an opportunity for spreading microbial contaminations under sub-optimal operating conditions. This study evaluated Salmonella inactivation and cross-contamination in a simulated washing process of cherry and grape tomatoes. Commercially harvested tomatoes and the associated inedible plant matter (debris) were differentially inoculated with kanamycin resistant (KanR) or rifampin resistant (RifR) Salmonella strains, and washed together with uninoculated tomatoes in simulated packinghouse dump tank (flume) wash water. Washing in chlorinated water resulted in significantly higher Salmonella reduction on tomatoes than on debris, achieving 2-3 log reduction on tomatoes and about 1 log reduction on debris. Cross-contamination by Salmonella on tomatoes was significantly reduced in the presence of 25-150 mg/L free chlorine, although sporadic cross-contamination on tomatoes was detected when tomatoes and debris were inoculated at high population density. The majority of the sporadic cross-contaminations originated from Salmonella inoculated on debris. These findings suggested that debris could be a potentially significant source of contamination during commercial tomato washing.

Interaction and multi-objective effects of multiple non-thermal treatments of sour cherry juice: pesticide removal, microbial inactivation, and quality preservation.

BACKGROUND: The consumption of pesticide-contaminated sour cherries as fruit or juice has become a major health concern, and so the search for alternative processing technologies, such as pulsed electric fields (PEF), ozone (O), and ultrasonication (US) has intensified. The objectives of this experimental study of sour cherry juice were fourfold: (1) to quantify the removal efficiency of new processing technologies (PEF, O, US), and their combinations, for the pesticides chlorpyrifos ethyl, τ-fluvalinate, cyprodinil, pyraclostrobin, and malathion; (2) to detect their impact on physical, bioactive, and sensory properties; (3) to determine their microbial inactivation levels for Escherichia coli O157:H7, Bacillus cereus, Pseudomonas syringae subs. Syringae, and Penicillum expansum; and (4) to jointly optimize multiple responses of physical, quality, and sensory properties, pesticides, and microbial inactivation. RESULTS: Except for all the O treatments, the physical, bioactive and sensory properties of sour cherry juice were not adversely affected by the treatments. The joint optimization suggested PEF1 (24.7 kV cm-1 for 327 µs), PEF2 (24.7 kV cm-1 for 655 µs), PEF2 + O + US, US, and PEF2 + O as the five best treatments. PEF2 + O + US best achieved both pesticide removal and microbial inactivation. CONCLUSION: PEF2 + O + US provided promising reductions in pesticide and microbial loads. © 2019 Society of Chemical Industry.

Genetic diversity of diazotrophs and total bacteria in the phyllosphere of Pyrus serotina, Prunus armeniaca, Prunus avium, and Vitis vinifera.

The phyllosphere, which supports a large number of microorganisms, represents the interface between the aboveground parts of plants and air. In this study, four nifH clone libraries were constructed from the phyllosphere of Pyrus serotina (L), Vitis vinifera (P), Prunus armeniaca (X), and Prunus avium (Y). Clones related to Skermanella (L, 12.1%; X, 15.6%; Y, 62.5%; P 70.8%), Bradyrhizobium (X, 2.1%; P, 15.1%; L, 63.7%), Erwinia (X, 68.8%), Pseudomonas (L, 3.3%; P, 7.6%), and Chroococcidiopsis (P, 0.9%; L, 4.4%, X; 5.2%, Y; 19.6%) were present at high percentages, highlighting their critical role in contributing nitrogen to the phyllosphere ecosystem. The 16S rDNA sequence analysis suggested that phyllosphere-associated bacteria were affiliated with a wide range of taxa, encompassing members from Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria, Deltaproteobacteria, Actinobacteria, Bacteroidetes, Firmicutes, Cyanobacteria, Tenericutes, and Deinococcus-Thermus. Additionally, the abundance of the nifH gene and 16S rDNA was assessed with quantitative PCR. The number of copies of nifH and 16S rDNA ranged from 1.14 × 103 to 1.49 × 104 and from 3.72 × 106 to 7.02 × 107 copies/g fresh leaf sample, respectively. In conclusion, our work sheds light on the microbial communities of the phyllosphere that are important for plant growth. Moreover, we observed a unique composition of nitrogen-fixing bacteria in each phyllosphere sample, suggesting the existence of specific interactions between these functional microorganism and plants, which may provide information or be a reference for the development of bacterial fertilizers.

Ethyl pyruvate (EP) suppressed post-harvest blue mold of sweet cherry fruit by inhibiting the growth of Penicillium oxalicum.

BACKGROUND: The shelf-life of fresh sweet cherry is relatively short due to fungal decay during post-harvest storage. To investigate the effect and the mechanism of action of ethyl pyruvate (EP) against blue mold of sweet cherry fruit caused by Penicillium oxalicum, the spores were treated with 25 mg L-1 EP. The spore germination rate of P. oxalicum, the integrities of the cell wall and plasma membrane, reactive oxygen species (ROS) and malondialdehyde (MDA) were evaluated. RESULTS: EP treatment significantly suppressed post-harvest blue mold of sweet cherry fruit. We found that the treatment of 25 mg L-1 EP significantly suppressed blue mold of post-harvest sweet cherry fruit by directly inhibiting germination and the mycelial growth of P. oxalicum. After co-inoculation with EP for 30 min, the spore germination rate of P. oxalicum was reduced by 83.5%. In addition, the pH of the EP solution was found to affect its antimicrobial activity. After treatment with EP, the cell surface structures of the spores of P. oxalicum were much more incomplete, and higher ROS and MDA values were recorded in the spores. CONCLUSIONS: The results suggested that EP treatment destroyed the integrities of the cell surface structures and caused oxidative damage of the spores of P. oxalicum. © 2019 Society of Chemical Industry.

Determination of Alternaria Mycotoxins in Fresh Sweet Cherries and Cherry-Based Products: Method Validation and Occurrence.

Sweet cherry is susceptible to disease caused by the Alternaria species and produces various Alternaria mycotoxins. Analytical methodologies based on solid-phase extraction (SPE) and LC-MS/MS to simultaneously determine five main Alternaria mycotoxins (tenuazonic acid, 1; alternariol, 2; alternariol methyl ether, 3; altenuene, 4; and tentoxin, 5) in fresh sweet cherries and cherry products were developed and validated. The limits of quantitation (LOQ) of the analytes ranged from 0.002-0.066 µg/kg. The method was successfully applied to 83 fresh cherry and cherry-related product samples. 1 and 5 were the predominant toxins with detection frequencies >50%, followed by 3 (42%), 2 (35%), and 4 (31%). Daily intakes of Alternaria mycotoxins via fresh sweet cherries were assessed preliminarily using the measured concentrations, and consumption data were obtained from a web-based dietary questionnaire ( n = 476). The maximum exposure of 1 and 3 were 4.6 and 16.7 times the threshold of the toxicological concern (TTC) value, respectively.

Comparative genomics of Pseudomonas syringae reveals convergent gene gain and loss associated with specialization onto cherry (Prunus avium).

Genome-wide analyses of the effector- and toxin-encoding genes were used to examine the phylogenetics and evolution of pathogenicity amongst diverse strains of Pseudomonas syringae causing bacterial canker of cherry (Prunus avium), including pathovars P. syringae pv morsprunorum (Psm) races 1 and 2, P. syringae pv syringae (Pss) and P. syringae pv avii. Phylogenetic analyses revealed Psm races and P. syringae pv avii clades were distinct and were each monophyletic, whereas cherry-pathogenic strains of Pss were interspersed amongst strains from other host species. A maximum likelihood approach was used to predict effectors associated with pathogenicity on cherry. Pss possesses a smaller repertoire of type III effectors but has more toxin biosynthesis clusters than Psm and P. syringae pv avii. Evolution of cherry pathogenicity was correlated with gain of genes such as hopAR1 and hopBB1 through putative phage transfer and horizontal transfer respectively. By contrast, loss of the avrPto/hopAB redundant effector group was observed in cherry-pathogenic clades. Ectopic expression of hopAB and hopC1 triggered the hypersensitive reaction in cherry leaves, confirming computational predictions. Cherry canker provides a fascinating example of convergent evolution of pathogenicity that is explained by the mix of effector and toxin repertoires acting on a common host.

Effect of ultrasound on some chemical and microbiological properties of sour cherry juice by response surface methodology.

In this study, it is aimed to determine effect of ultrasonication on some chemical and microbiological properties of sour cherry juice by response surface methodology, since ultrasound is known as an alternative method for thermal food processing. Sour cherry juice was sonicated at varying amplitude levels (50, 75, 100%); moderate temperatures (20, 30, 40 ℃); and treatment times of 2, 6, 10 min at a constant frequency of 20 kHz. Different ultrasonication amplitudes, temperatures, and times had no significant effect on pH,°Bx, and titratable acidity. A significant increase in total monomeric anthocyanins was observed as the amplitude level and temperature increased (p < 0.01). An increase in the total phenolics was also obtained as the temperature increased (p < 0.05). The effect of amplitude level on antioxidant capacity of sour cherry juice was also found significant (p < 0.05). Color parameters (L*, a*, b*, C, h) generally increased by increasing temperature, amplitude level, and treatment time. It was determined that Escherichia coli O157:H7 significantly affected by temperature and treatment time (p < 0.05).

Antifungal effect of volatile organic compounds produced by Bacillus amyloliquefaciens CPA-8 against fruit pathogen decays of cherry.

The present work focuses on the antifungal effect of volatile organic compounds (VOCs) produced by Bacillus amyloliquefaciens CPA-8 against Monilinia laxa, M. fructicola and Botrytis cinera, three postharvest fruit pathogens of sweet cherry fruit. VOCs were evaluated with a double petri dish assay against mycelial and colony growth of target pathogens. For this purpose, CPA-8 was grown on different media and cultured for 24 and 48 h at 30 °C before assays. Data showed that mycelial growth inhibition was higher when CPA-8 was grown on Tryptone Soya Agar (TSA) while no differences were generally observed when CPA-8 was cultured for either, 24 and 48 h. Moreover, no effects were observed on colony growth. The main volatile compounds emitted by CPA-8 were identified by solid-phase microextraction (SPME)-gas chromatography as 1,3 pentadiene, acetoin (3-hydroxy-2-butanone) and thiophene. Pure compounds were also tested in vitro on mycelial growth inhibition and their EC50 values against the three pathogens were estimated. Thiophene was the most effective VOC, showing more than 82% suppression of mycelial growth at the highest concentration (1.35 µL/mL headspace) and EC50 values ranging from 0.06 to 6.67 µL/mL headspace. Finally, the effectiveness of thiophene and CPA-8 VOCs was evaluated against artificially inoculated cherry fruits. Among the target pathogens, M. fructicola was clearly controlled by CPA-8 with less than 25% of rotten fruits compared to the control (65% disease incidence) and for all pathogens, less than 37.5% of CPA-8 treated decayed fruits produced spores (disease sporulation). Otherwise, pure thiophene showed no effect against any pathogen on disease incidence and disease sporulation. The results indicated that VOCs produced by B. amyloliquefaciens CPA-8 could develop an additive antifungal effect against postharvest fruit pathogens on stone fruit.

PacMYBA, a sweet cherry R2R3-MYB transcription factor, is a positive regulator of salt stress tolerance and pathogen resistance.

Plant R2R3-MYB transcription factors play crucial roles in stress responses. We previously isolated a R2R3-MYB homolog from sweet cherry cv. Hong Deng, designated PacMYBA (GenBank accession No. KF974774). To explore the role of PacMYBA in the plant stress response, we heterologously expressed PacMYBA in transgenic Arabidopsis thaliana plants. In a previous study, we demonstrated that PacMYBA is mainly localized to the nucleus and could be induced by abscisic acid (ABA). Analysis of the promoter sequence of PacMYBA revealed that it contains several stress-related cis-elements. QPCR results showed that PacMYBA is induced by salt, salicylic (SA), and jasmonic acid (JA) in sweet cherry leaves. Transgenic Arabidopsis plants heterologously expressing PacMYBA exhibited enhanced salt-tolerance and increased resistance to Pseudomonas syringe pv. tomato (Pst) DC3000 infection. Overexpression of PacMYBA decreased the osmotic potential (OP), increased the free proline content, and increased the peroxidase content in transgenic Arabidopsis plants. Furthermore, overexpression of PacMYBA also affected the expression levels of salt stress- and pathogen defense-related genes in the transgenic plants. These results indicate that PacMYBA is a positive regulator of salt stress tolerance and pathogen resistance.

Combined effect of antagonistic yeast and modified atmosphere to control Penicillium expansum infection in sweet cherries cv. Ambrunés.

Fruit decay caused by pathogenic moulds is a major concern in the postharvest quality and shelf life of fruit. Blue mould decay is caused by Penicillium expansum (P. expansum) and is one of the most important postharvest diseases in cherries (Prunus avium L.). Synthetic fungicides are the main medium used to control pathogenic moulds. However, alternative approaches are available for developing safer technologies to control postharvest disease. An integrated approach that combines biological control, using antagonistic yeasts and modified atmosphere packaging (MAP) with cold storage is a promising alternative to synthetic fungicide treatment. In this work, two microperforated films (M10 and M50) and two antagonistic yeast strains (Hanseniaspora opuntiae L479 and Metschnikowia pulcherrima L672) were evaluated for their effectiveness to control the development of P. expansum in wounded cherries stored at 1°C. Results showed that the microperforated films had fungistatic effects, particularly M50, due to the level of CO2 achieved (mean CO2 of 11.2kPa at 35days), and the decrease in disease severity. Antagonistic yeasts, particularly Metschnikowia pulcherrima L672, delayed the development of P. expansum and decreased disease incidence and severity. The combination of MAP and antagonistic yeasts was the most effective approach to control P. expansum, during cold storage.

Characterization of community structure of culturable endophytic fungi in sweet cherry composite trees and their growth-retarding effect against pathogens.

Endophytic fungi have the potential to protect their host plants in stress situations. Characterizing the ecology and complex interaction between these endophytes and their host plants is therefore of great practical importance, particularly in horticultural plants. Among horticultural plants, fruit trees form a special category because of their longevity and because they are composites of rootstock and scion, which often belong to different plant species. Here we present the first characterization of culturable endophytic fungal community of sweet cherry. Samples from the Hungarian cultivar 'Petrus' grafted on 11 different rootstocks were collected in autumn and in spring in a bearing orchard and the dependence of colonization rate and endophyte diversity on rootstock, organ and season was analysed. On the basis of their ITS sequences 26 fungal operational taxonomic units were identified at least down to the genus level. The dominant genus, comprising more than 50% of all isolates, was Alternaria, followed by different Fusarium and Epicoccum species. We observed some organ-specificity amongst endophytes, and organs showed more sizeable differences in colonization rates and endophyte diversity than rootstocks. Most dynamic endophyte populations, strongly influenced by environmental conditions and crop management, were observed in leaves. The potential of selected endophytes to confer protection against Monilinia laxa was also analysed and 7 isolates were found to inhibit the growth of this pathogen in vitro.

Pseudomonas cerasi sp. nov. (non Griffin, 1911) isolated from diseased tissue of cherry.

Eight isolates of Gram-negative fluorescent bacteria (58(T), 122, 374, 791, 963, 966, 970a and 1021) were obtained from diseased tissue of cherry trees from different regions of Poland. The symptoms resembled those of bacterial canker. Based on an analysis of 16S rDNA sequences the isolates shared the highest over 99.9% similarity with Pseudomonas ficuserectae JCM 2400(T) and P. congelans DSM 14939(T). Phylogenetic analysis using housekeeping genes gyrB, rpoD and rpoB revealed that they form a separate cluster and confirmed their closest relation to P. syringae NCPPB 281(T) and P. congelans LMG 21466(T). DNA-DNA hybridization between the cherry isolate 58(T) and the type strains of these two closely related species revealed relatedness values of 58.2% and 41.9%, respectively. This was further supported by Average Nucleotide Identity (ANIb) and Genome-to-Genome Distance (GGDC) between the whole genome sequences of strain LMG 28609(T) and closely related Pseudomonas species. The major cellular fatty acids are 16:0 and summed feature 3 (16:1 ω7c/15:0 iso 2OH). Phenotypic characteristics differentiated the novel isolates from other closely related species. The G+C content of the genomic DNA of strain 58(T) was 59%. The diversity was proved by PCR MP and BOX PCR, eliminating the possibility that they constitute a clonal population. Based on the evidence of this polyphasic taxonomic study the eight strains are considered to represent a novel species of the genus Pseudomonas for which the name P. cerasi sp. nov. (non Griffin, 1911) is proposed. The type strain of this species is 58(T) (=LMG 28609(T)=CFBP 8305(T)).

Pararhizobium polonicum sp. nov. isolated from tumors on stone fruit rootstocks.

Five Gram-negative, rod-shaped, non-spore-forming bacteria were isolated from galls on different stone fruit rootstocks in Poland: strains F5.1(T) and F5.3 from Prunus avium F12/1, strains CP3.5 and CP17.2.1 from Prunus avium and strain AL5.1.8 from Prunus cerasifera. On the basis of 16S rDNA phylogeny, the strains cluster together and belong to the genus Pararhizobium with type strain of Pararhizobium herbae (99.6-99.8%) as their closest relative. Phylogenetic analysis of the novel strains using housekeeping genes atpD, recA and rpoB revealed their distinct position separate from other known Rhizobium species and confirmed their relation to P. herbae. DNA-DNA hybridization of strains F5.1(T), with the type strain of P. herbae LMG 25718(T) and Pararhizobium giardinii R-4385(T) revealed 28.3% and 27.9% of DNA-DNA relatedness, respectively. Phenotypic and physiological properties differentiate the novel isolates from other closely related species. On the basis of the results obtained, the five isolates are considered to represent a novel species of the genus Pararhizobium, for which the name Pararhizobium polonicum sp. nov. (type strain F5.1(T)=LMG 28610(T)=CFBP 8359(T)) is proposed.

Effect of phytosanitary irradiation and methyl bromide fumigation on the physical, sensory, and microbiological quality of blueberries and sweet cherries.

BACKGROUND: The objective of this study was to determine whether irradiation could serve as a suitable phytosanitary treatment alternative to methyl bromide (MB) fumigation for blueberries and sweet cherry and also to determine the effect of phytosanitary irradiation treatment on survival of Salmonella spp. and Listeria monocytogenes on these fruit. 'Bluecrop' blueberries (Vaccinium corymbosum) and 'Sweetheart' cherries (Prunus avium) were irradiated at 0.4 kGy or fumigated with methyl bromide and evaluated for quality attributes during storage. RESULTS: Irradiation caused an immediate decrease in firmness of both fruit without further significant change during storage. Fumigated fruit, in contrast, softened by 11-14% during storage. Irradiation did not adversely affect blueberry and cherry shelf-life. MB fumigation did not impact blueberry and cherry quality attributes initially; however, fumigated fruit exhibited greater damage and mold growth than the control and irradiated samples during storage. Irradiation at 400 Gy resulted in a ∼1 log CFU g(-1) reduction in Salmonella spp. and Listeria monocytogenes counts, indicating that this treatment cannot significantly enhance safety. CONCLUSION: This study indicates that irradiation at a target dose of 0.4 kGy for phytosanitary treatment does not negatively impact blueberry and cherry quality and can serve as an alternative to methyl bromide fumigation. © 2016 Society of Chemical Industry.