Cherry picking by pseudomonads: After a century of research on canker,genomics provides insights into the evolution of pathogenicity towards stone fruits

Bacterial canker disease is a major limiting factor in the growing of cherry and other Prunus species worldwide. At least five distinct clades within the bacterial species complex Pseudomonas syringae are known to be causal agents of the disease. The different pathogens commonly coexist in the field. Reducing canker is a challenging prospect as the efficacy of chemical controls and host resistance may vary against each of the diverse clades involved. Genomic analysis has revealed that the pathogens use a variable repertoire of virulence factors to cause the disease. Significantly, strains of P. syringae pv. syringae possess more genes for toxin biosynthesis and fewer encoding type III effector proteins. There is also a shared pool of key effector genes present on mobile elements such as plasmids and prophages that may have roles in virulence. By contrast, there is evidence that absence or truncation of certain effector genes, such as hopAB, is characteristic of cherry pathogens. Here we highlight how recent research, underpinned by the earlier epidemiological studies, is allowing significant progress in our understanding of the canker pathogens. This fundamental knowledge, combined with emerging insights into host genetics, provides the groundwork for development of precise control measures and informed approaches to breed for disease resistance.     

Identification and Discrimination of Pseudomonas syringae Isolates from Wild Cherry in England

A survey of wild cherry (Prunus avium) woodland plantations and nurseries was carried out in 2000/01. Trees with symptoms of bacterial canker were found in 20 of the 24 plantations visited and in three of seven nurseries. Fifty-four Pseudomonas syringae isolates from wild cherry together with 22 representative isolates from sweet cherry and 13 isolates from other Prunus spp., pear and lilac were characterised by physiological, biochemical, serological and pathogenicity tests. Isolates from wild cherry were predominantly P. syringae pv. syringae (Pss), but P. syringae pv. morsprunorum (Psm) races 1 and 2 were also found. Physiological and biochemical tests discriminated Psm races 1 and 2 from other P. syringae isolates. Agglutination and indirect-enzyme-linked immunosorbent assay tests with three different antisera showed that Psm race 1 and race 2 were very uniform and indicated high variability amongst other P. syringae isolates. However, pathogenic Pss isolates could not be distinguished from non-pathogenic isolates of P. syringae on the basis of physiological, biochemical or serological tests. Pathogenicity tests on rooted lilac plants and on micropropagated plantlets of lilac and two wild cherry clones differentiated Pss and Psm isolates and demonstrated a range of aggressiveness amongst Pss isolates. Serological tests could be used as an alternative to the classical physiological and biochemical tests to increase the speed of detection and discrimination of isolates, but pathogenicity tests are still necessary to discriminate the pathogenic Pss isolates.     

Discrimination of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> isolates from sweet and wild cherry using rep-PCR

Bacterial canker is one of the most important diseases of cherry (Prunus avium). This disease can be caused by two pathovars of Pseudomonas syringae: pv. morsprunorum and pv. syringae. Repetitive DNA polymerase chain reaction-based fingerprinting (rep-PCR) was investigated as a method to distinguish pathovars, races and isolates of P. syringae from sweet and wild cherry. After amplification of total genomic DNA from 87 isolates using the REP (repetitive extragenic palindromic), ERIC (enterobacterial repetitive intergenic consensus) and BOX primers, followed by agarose gel electrophoresis, groups of isolates showed specific patterns of PCR products. Pseudomonas syringae pv. syringae isolates were highly variable. The differences amongst the fingerprints of P. syringae pv. morsprunorum race 1 isolates were small. The patterns of P. syringae pv. morsprunorum race 2 isolates were also very uniform, with one exception, and distinct from the race 1 isolates. rep-PCR is a rapid and simple method to identify isolates of the two races of P. syringae pv. morsprunorum; this method can also assist in the identification of P. syringae pv. syringae isolates, although it cannot replace inoculation on susceptible hosts such as cherry and lilac.     

Pseudomonas syringae pv. avii (pv. nov.), the Causal Agent of Bacterial Canker of Wild Cherries (Prunus avium) in France

Bacterial strains isolated from cankers of wild cherry trees (Prunus avium) in France were characterized using numerical taxonomy of biochemical tests, DNA–DNA hybridization, repeat sequence primed-PCR (rep-PCR) based on REP, ERIC and BOX sequences, heteroduplex mobility assay (HMA) of internal transcribed spacer (ITS) as well as pathogenicity on wild cherry trees and other species of Prunus. They were compared to reference strains of Pseudomonas syringae pathovars isolated from wild and sweet cherry and various host plants. Wild cherry strains were closely related to P. syringae (sensu lato) in LOPAT group Ia (+ - - - +). Wild cherry strains were pathogenic to wild cherry trees and produced symptoms similar to those observed in orchards. They were pathogenic also, but at a lesser extent, to sweet cherry trees (cv. Napoléon). The wild cherry strains were collected from five different areas in France and appeared to constitute a very homogeneous group. They showed an homogenous profile of a biochemical and physiological characteristics. They were closely related by DNA–DNA hybridization and belonged to genomospecies 3 `tomato'. Rep-PCR showed that wild cherry strains constitute a tight group distinct from P. s. pv. morsprunorum races 1 and 2 and from other P. syringae pathovars. HMA profiles indicated that the ITS of all wild cherry strains were identical but different from P. s. pv. persicae strains since the two heteroduplex bands with reduced mobility were generated by hybridization with the P. s. pv. persicae pathotype strain CFBP 1573. The 8 genomospecies of Gardan et al. (1999) have not been converted into formal species as they cannot be differentiated by biochemical tests. Therefore, the pathovar system within P. syringae was currently used. P. syringae pv. avii is proposed for this bacterium causing a wild cherry bacterial canker and strain CFBP 3846 (NCPPB 4290, ICMP 14479) is designated as the pathotype.     

Characterization of the pathogenicity of strains of Pseudomonas syringae towards cherry and plum

Bacterial canker is a major disease of Prunus avium (cherry), Prunus domestica (plum) and other stone fruits. It is caused by pathovars within the Pseudomonas syringae species complex including P. syringae pv. morsprunorum (Psm) race 1 (R1), Psm race 2 (R2) and P. syringae pv. syringae (Pss). Psm R1 and Psm R2 were originally designated as the same pathovar; however, phylogenetic analysis revealed them to be distantly related, falling into phylogroups 3 and 1, respectively. This study characterized the pathogenicity of 18 newly genome‐sequenced P. syringae strains on cherry and plum, in the field and laboratory. The field experiment confirmed that the cherry cultivar Merton Glory exhibited a broad resistance to all clades. Psm R1 contained strains with differential specificity on cherry and plum. The ability of tractable laboratory‐based assays to reproduce assessments on whole trees was examined. Good correlations were achieved with assays using cut shoots or leaves, although only the cut shoot assay was able to reliably discriminate cultivar differences seen in the field. Measuring bacterial multiplication in detached leaves differentiated pathogens from nonpathogens and was therefore suitable for routine testing. In cherry leaves, symptom appearance discriminated Psm races from nonpathogens, which triggered a hypersensitive reaction. Pathogenic strains of Pss rapidly induced disease lesions in all tissues and exhibited a more necrotrophic lifestyle than hemibiotrophic Psm. This in‐depth study of pathogenic interactions, identification of host resistance and optimization of laboratory assays provides a framework for future genetic dissection of host–pathogen interactions in the canker disease.     

Assessment of the inheritance of resistance and tolerance in cherry (Prunus sp.) to Blumeriella jaapii,the causal agent of cherry leaf spot

Cherry leaf spot (CLS), caused by Blumeriella jaapii, is a serious fungal disease of sour cherry (Prunus cerasus). Cultivar Montmorency, the major cultivar grown in the United States, is highly susceptible to CLS. As many as 10 fungicide sprays can be required each growing season to combat this disease; therefore, developing CLS‐resistant cultivars is a top breeding priority. Germplasm previously reported to be resistant or tolerant to CLS was acquired and incorporated into the sour cherry breeding programme at Michigan State University (MSU) and included three cherry species: sour cherry, sweet cherry (P. avium), and the wild species P. canescens. This study aimed to: (i) compare the CLS disease progression profile of the susceptible cultivar Montmorency with those of the resistant and tolerant germplasm; and (ii) gain an understanding of the inheritance of these resistance and tolerance traits by evaluating the host response of progeny individuals belonging to families derived from this germplasm. Significant differences were observed between the susceptible Montmorency and the tolerant and resistant accessions in their response to CLS and its progression during the growing season. Evaluation of the CLS host responses of progeny individuals derived from this germplasm supported a dominant two‐gene model for P. canescens‐derived resistance and a recessive gene model for sweet cherry‐derived tolerance. These insights into disease progression and trait inheritance improve the efficiency and potential success of breeding sour cherry cultivars with durable resistance to CLS.     

Bacteria from four phylogroups of the Pseudomonas syringae complex can cause bacterial canker of apricot

Pseudomonas syringae is described as a species complex, containing P. syringae-related species classified into 13 phylogroups and 23 clades. Pseudomonas syringae is one of the main pathogens of fruit trees, affecting nut trees, hazelnut and kiwi, pome and stone fruits. Bacterial canker of apricots is an important disease in regions of production with cold winters and conducive soils. This work characterizes the bacteria able to induce canker in apricots isolated in different French orchards. Bacteria from four phylogroups were able to induce canker. The pathogenicity to apricot was not linked to the pathogenicity to the three herbaceous species and cherry fruits tested, and was not always related to hypersensitive reaction on tobacco and ice nucleation activity. Bacteria pathogenic to apricot belong to phylogroups 01, 02, 03 and 07. The bacteria of phylogroups 01a and 07a (Pseudomonas viridiflava) characterized in this work have not previously been described as pathogenic to apricot.     

Molecular data from up to 130-year-old herbarium specimens do not support the presence of cherry powdery mildew in Australia

A strain of Podosphaera clandestina has been highlighted as a priority pest threat to the Australian cherry industry. Australia currently has no records of powdery mildew on cherry (Prunus avium). P. clandestina is reported to cause disease on a range of Rosaceae genera including Crataegus and Prunus; in Australia, P. clandestina has only been recorded on Crataegus. A recent species revision identified Podosphaera cerasi on P. avium as a separate species from P. clandestina. Therefore, a revision of which powdery mildew species is present in Australia on Crataegus is required to inform Australian plant biosecurity. Reference collection specimens from the Victorian Plant Pathology Herbarium (VPRI) recorded as Podosphaera spp. collected between 1889 to 2008 on cherry and three other host plant genera from Australia and overseas were sampled for DNA extraction and next-generation sequencing (NGS). Sequence data from preserved specimens were successfully mapped to internal transcribed spacer (ITS) sequences of P. clandestina in the strict sense, P. cerasi, and Podosphaera prunicola, and chloroplast matK sequences were used to identify plant hosts. Australian specimens on Crataegus hosts were P. clandestina in the strict sense and specimens on Prunus from the USA were identified as P. cerasi and P. prunicola. The outcome of this study confirmed the powdery mildew on Australian Crataegus specimens to be P. clandestina and none of the cherry powdery mildews (Podosphaera pruni-avium, P. cerasi, or P. prunicola) are present on Australian specimens in the VPRI collection, which suggests they are not present in Australia.     

Evaluation of cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>) cultivars grown in Eastern Europe and progress in breeding for resistance to angular leaf spot (<Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">lachrymans</Emphasis>)

Increased occurrence of cucumber angular leaf spot, Pseudomonas syringae pv. lachrymans, has caused significant losses in cucumber, Cucumis sativus, yield in Poland in recent years. These losses necessitated evaluation of the level of resistance in cucumber cultivars of mainly Polish breeding, cultivated in Eastern Europe, and initiation of a breeding programme for resistance to this disease. Screening for resistance was performed on 84 cucumber accessions under growth chamber conditions using a highly aggressive strain of P. syringae pv. lachrymans. Most of the screened accessions were either susceptible or displayed intermediate resistance. The screening resulted in the identification of five F₁ hybrid cultivars moderately resistant to angular leaf spot. The identified F₁ hybrids were self-pollinated up to the F₄ generation. Individuals resistant to angular leaf spot were identified. These individuals can be used as a source of resistance to angular leaf spot in future breeding efforts.     

Sources of resistance to Pseudocercospora fijiensis,the cause of black Sigatoka in banana

Black Sigatoka, caused by Pseudocercospora fijiensis, is one of the most devastating diseases of banana. In commercial banana-growing systems, black Sigatoka is primarily managed by fungicides. This mode of disease management is not feasible for resource-limited smallholder farmers. Therefore, bananas resistant to P. fijiensis provide a practical solution for managing the disease, especially under smallholder farming systems. Most banana and plantain hybrids with resistance to P. fijiensis were developed using few sources of resistance, which include Calcutta 4 and Pisang Lilin. To broaden the pool of resistance sources to P. fijiensis, 95 banana accessions were evaluated under field conditions in Sendusu, Uganda. Eleven accessions were resistant to P. fijiensis. Black Sigatoka symptoms did not progress past Stage 2 (narrow brown streaks) in the diploid accessions Pahang (AA), Pisang KRA (AA), Malaccensis 0074 (AA), Long Tavoy (AA), M.A. Truncata (AA), Tani (BB), and Balbisiana (BB), a response similar to the resistant control Calcutta 4. These accessions are potential sources of P. fijiensis resistance and banana breeding programmes can use them to broaden the genetic base for resistance to P. fijiensis.     

Over-expression of the Arabidopsis <Emphasis Type="BoldItalic">NPR1</Emphasis> gene in citrus increases resistance to citrus canker

Citrus canker, caused by the bacterial pathogen Xanthomonas citri subsp. citri (Xcc), is a serious leaf and fruit spotting disease affecting many important citrus cultivars including grapefruit and certain sweet oranges. Currently, efficacious and economical disease control measures for highly susceptible citrus cultivars are lacking. Development of commercial cultivars with greater field resistance to citrus canker is the optimum strategy for effective disease management. In this study, we generated transgenic ‘Duncan’ grapefruit (DG) and ‘Hamlin’ sweet orange (Ham) expressing the Arabidopsis NPR1 gene (AtNPR1), which is a key positive regulator of the long-lasting broad-spectrum resistance known as systemic acquired resistance (SAR). Our results indicate that over-expression of AtNPR1 in citrus increases resistance to citrus canker and that the resistance is related with the expression levels of AtNPR1 in the transgenic plants. The line (DG 42-2) with the highest expression level of AtNPR1 was also the most resistant, which developed significant fewer lesions accompanied by a ten-fold reduction in Xcc population. The lesions developed on DG 42-2 were smaller and darker than those on the control and lacked callus formation. These lesion phenotypes resemble those on canker resistant kumquats and canker susceptible citrus trees treated with SAR-inducing compounds. Therefore, over-expression of AtNPR1 in citrus is a promising approach for development of more resistant cultivars to citrus canker.     

Effects of temperature,free moisture duration and inoculum concentration on infection of sweet cherry by<Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv.<Emphasis Type="Italic">syringae</Emphasis>

The effects of temperature, free moisture duration and inoculum concentration on infection caused byPseudomonas syringae pv.syringae (Pss), on sweet cherry (Prunus avium) were investigated. Epiphytic populations ofPss are an important source of inoculum for bacterial canker and it has been demonstrated that a cyclic pattern exists during the year, from undetectable during the warm and dry periods to large populations following cool and wet periods. The effects of temperature and inoculum concentration on the infection caused byPss on immature fruits and 1-yr-old twigs were significant (P<0.001). Fruit and twig infection increased linearly in proportion to the logarithm ofPss when bacterial concentrations were higher than 10³ cfu ml⁻¹ and temperatures were between 5 and 20°C. Regardless of the inoculum concentration and the free moisture duration, fruit and twig infection was either absent or low at 5°C but it increased linearly as temperature increased from 5 to 20°C. Growth ratein vitro was very slow (0.03–0.04 cfu h⁻¹) at 5°C and fast (0.21–0.23 cfu h⁻¹) at 20°C. Therefore, it is possible that multiplication of the epiphytic populations may be significantly reduced in the field with air temperatures below 5°C. A significant (P<0.001) effect of free moisture was obtained only when a low inoculum concentration (10³ cfu ml⁻¹) was used, and a significant linear response between free moisture and disease incidence was obtained only at 10°C. An apparent threshold population ofPss higher than 10³ cfu ml⁻¹ was needed to infect immature fruits and 1-yr-old twigs of sweet cherry. http://www.phytoparasitica.org posting July 10, 2002.     

Genetic diversity and pathogenicity of Pseudomonas syringae pv. aptata isolated from sugar beet

Pseudomonas syringae pv. aptata is the causal agent of bacterial leaf spot disease of sugar beet (Beta vulgaris). During 2013, 250 samples were collected from leaf lesions with typical symptoms of bacterial leaf spot in commercial fields of sugar beet in Serbia, and 104 isolates of P. syringae pv. aptata were obtained. Identification and characterization was performed using biochemical, molecular and pathogenicity tests. Identification included LOPAT tests and positive reactions using primers Papt2F and Papt1R specific for P. syringae pv. aptata. Repetitive (rep) sequence‐based PCR typing with ERIC, REP and BOX primers revealed high genetic variability among isolates and distinguished 25 groups of different fingerprinting profiles. Pulse‐field gel electrophoresis (PFGE) and multilocus sequence analysis (MLSA) of representative isolates showed higher genetic variability than in rep‐PCR analysis and distinguished three and four major genetic clusters, respectively. A pathogenicity test performed with 25 representative isolates on four cultivars of sugar beet confirmed the occurrence of leaf spot disease and showed correlation between the most aggressive isolates and the genetic clusters obtained in MLSA. All these findings point to the existence of several lines of P. syringae pv. aptata infection in Serbia that are genetically and pathologically different.     

Influence of rootstock on the susceptibility of sweet cherry scions to bacterial canker, caused by Pseudomonas syringae pvs morsprunorum and syringae

In a field trial to determine whether the rootstock influenced the susceptibility of cherry cultivars to bacterial canker three cultivars (Napoleon, Roundel and JI 14039), each grafted on two rootstocks (F 12/1 and Colt), were subjected to natural infectionand to inoculation with three bacterial canker pathogens (Pseudomonas syringae pv. morsprunorum races 1 and 2 and P. syringae pv. syringae). Inoculations were made through leaf scars and through wounds. The high susceptibility of Napoleon and high resistance of JI 14039 were confirmed. Napoleon was more susceptible to inoculation through branches when on F12/1 than when on Colt but the reverse was true for leaf scar inoculations. JI14039 was more susceptible to race 1 inoculated through leaf scars when grown on F12/1 than when on Colt. No rootstock/scion interaction was detected with Roundel.
The complexity of the relationships between the pseudomonad pathogens and their cherry hosts is briefly discussed.     

Nucleotide Sequence and Organization of Copper Resistance Genes from Pseudomonas syringae pv. actinidiae

Copper-containing bactericides have been used to control bacterial canker of kiwifruit, caused by Pseudomonas syringae pv. actinidiae. However, the efficacy of copper has been reduced by the occurrence of copper-resistant strains. Analysis of the DNA sequence of a cluster region containing the copper-resistance genes from P. syringae pv. actinidiae suggested the presence of three possible different systems for copper resistance: copper-trapping, copper-efflux and copper-transport systems. Transposon insertional inactivation analysis indicated that the copper-trapping system was essential for copper resistance.     

Pseudomonas syringae pv. syringae induces systemic resistance to Pyricularia oryzae in rice

Systemic resistance was induced to Pyricularia aryzae in rice by inoculation of the first leaf with the hypersensitive response causing bacterium, Pseudomonas syringae pv. syringae. Lesions caused by Pyricularia oryzae were decreased in number and size by 85% and 50%, respectively, in systemically protected leaves. Increased resistance was associated with the deposition of a dark brown material around sites of Pyricularia oryzae infection. The systemically acquired resistance was not associated with an increase in the activities of phenylalanine ammonia lyase, coniferyl alcohol dehydrogenase, peroxidase, β-1, 3-glucanase or chitinase after the challenge inocculation with Pyricularia oryzae. The levels of these enzymes were elevated by local exposure to Pseudomonas syringae pv. syringae or Pyricularia oryzae, but were not systemically induced. These data suggest that the physiological changes which occur during induced resistance in rice are different from those correlated with induced resistance in tobacco or cucumber.     

Reduction of Bacterial Speck (Pseudomonas syringae pv. tomato) of Tomato by Combined Treatments of Plant Growth-promoting Bacterium, Azospirillum brasilense, Streptomycin Sulfate, and Chemo-thermal Seed Treatment

Inoculation of tomato seeds with the plant growth-promoting bacterium Azospirillum brasilense, or spraying tomato foliage with A. brasilense, streptomycin sulfate, or commercial copper bactericides, separately, before or after inoculation with Pseudomonas syringae pv. tomato, the casual agent of bacterial speck of tomato, had no lasting effect on disease severity or on plant height and dry weight. Seed inoculation with A. brasilense combined with a single streptomycin foliar treatment and two foliar bactericide applications at 5-day intervals (a third or less of the recommended commercial dose) reduced disease severity in tomato seedlings by over 90% after 4 weeks, and significantly slowed disease development under mist conditions. A. brasilense did not induce significant systemic resistance against the pathogen although the level of salicylic acid increased in inoculated plants. Treatment of tomato seeds that were artificially inoculated with P. syringae pv. tomato, with a combination of mild chemo-thermal treatment, A. brasilense seed inoculation, and later, a single foliar application of a copper bactericide, nearly eliminated bacterial leaf speck even when the plants were grown under mist for 6 weeks. This study shows that a combination of otherwise ineffective disease management tactics, when applied in concert, can reduce bacterial speck intensity in tomatoes under mist conditions.     

Genetic analyses of Pseudomonas syringae isolates from Belgian fruit orchards reveal genetic variability and isolate-host relationships within the pathovar syringae, and help identify both races of the pathovar morsprunorum

A collection of Pseudomonas syringae and viridiflava isolates was established between 1993 and 2002 from diseased organs sampled from 36 pear, plum and cherry orchards in Belgium. Among the 356 isolates investigated in this study, phytotoxin, siderophore and classical microbiology tests, as well as the genetical methods REP-, ERIC- and BOX- (collectively, rep-) and IS50-PCR, enabled identification to be made of 280 isolates as P. syringae pv. syringae (Pss), 41 isolates as P. syringae pv. morsprunorum (Psm) race 1, 12 isolates as Psm race 2, three isolates as P. viridiflava and 20 isolates as unclassified P. syringae. The rep-PCR methods, particularly BOX-PCR, proved to be useful for identifying the Psm race 1 and Psm race 2 isolates. The latter race was frequent on sour cherry in Belgium. Combined genetic results confirmed homogeneities in the pvs avii, and morsprunorum race 1 and race 2 and high diversity in the pv. syringae. In the pv. syringae, homogeneous genetic groups consistently found on the same hosts (pear, cherry or plum) were observed. Pathogenicity on lilac was sometimes variable among Pss isolates from the same genetic group; also, some Psm race 2 and unclassified P. syringae isolates were pathogenic to lilac. In the BOX analyses, four patterns included 100% of the toxic lipodepsipeptide (TLP)-producing Pss isolates pathogenic to lilac. Many TLP-producing Pss isolates non-pathogenic to lilac and the TLP-non-producing Pss isolates were classified differently. Pseudomonas syringae isolates that differed from known fruit pathogens were observed in pear, sour cherry and plum orchards in Belgium.     

<Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> occurs asymptomatically on sweet cherry leaves

Leaves of sweet cherry, exposed to either paraquat or freezing to quickly senesce the leaf tissue, were incubated in about 100% RH at 25°C for 6 d. Sporulating colonies of Colletotrichum acutatum, the cause of anthracnose, developed on up to 100% of the paraquat-treated and frozen leaves, and on none of the untreated controls. Number of leaves and leaf area containing C. acutatum on naturally infected leaves increased over time from May to September. Mean incidence of C. acutatum on leaf blades on fruit spurs and vegetative shoots from eight orchard/year samplings were 41 and 33%, respectively. Secondary conidiation (formation of short hyphae and new conidia) from conidia applied to detached leaves took place 6 h after inoculation, but only up to 3% of the conidia formed new conidia. It may be concluded that asymptomatic sweet cherry leaves frequently host C. acutatum and may be a potential inoculum source for cherry fruit.     

The use of PCR melting profile for typing of <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> isolates from stone fruit trees

Of thirty fluorescent Pseudomonas isolates originating from symptomatic tissues of sweet (Prunus avium) and sour cherry (Prunus cerasus), plum (Prunus domestica), peach (Prunus persica) and apricot (Prunus armeniaca), 23 were identified as P. syringae using LOPAT tests. Further characterization of those isolates by GATTa and L-lactate utilization tests showed that 10 of them belonged to race 1, six to race 2 of P. syringae pv. morsprunorum (Psm) and six other isolates were identified as pathovar syringae (Pss). One isolate (791) was determined as atypical. Phenotypic determination and genetic analysis of studied isolates for toxin production revealed that isolates of Pss produced syringomycin, 3 Psm race 1 produced coronatine and 6 Psm race 2 produced yersiniabactin. Genetic diversity of all isolates was evaluated with the PCR melting profile (PCR MP) method. A dendrogram constructed with PCR MP patterns showed positive correlation with phenotypically distinguished pathovars. Isolates of Psm races 1 and 2 formed distinct, tight clusters, whereas Pss isolates were more heterogeneous. Isolate 791 was placed within Pss isolates. Bacteria identified as Pss caused more severe symptoms on immature cherry fruits compared to Psm, which corresponded to determined pathovars and races.