Syringae Strains Research Articles

A qPCR assay for specific detection of Pseudomonas cannabina pv. alisalensis.

Pseudomonas cannabina pv. alisalensis is a gram-negative bacterium that causes bacterial leaf blight in Brassica crops, an important disease that could bring severe damage to the host plants. The aim of this study was to develop a tool that can reliably and accurately quantify P. cannabina pv. alisalensis and distinguish it from other closely related bacterial pathogens. Two species and six pathovars of Pseudomonas were tested: three pathovars, P. syringae pv. coriandricola, P. syringae pv. philadelphi, and P. syringae strains from Vicia faba were found or confirmed to be members of P. cannabina based on the multi-locus sequence analysis and rep-PCR results. The qPCR assay was evaluated for specificity and examined for detection limit in pure bacterial cells and bacteria-spiked plant samples. The assay was applied in monitoring the quantities of the P. cannabina pv. alisalensis DNA over time in inoculated turnip green leaves. As results, the newly developed qPCR assay detected the target DNA in P. cannabina pv. alisalensis suspension as low as 100 CFU/ml and did not detect any of the nontarget bacteria. The qPCR assay detected P. cannabina pv. alisalensis in all the inoculated samples at least 5 days before the symptoms became visible; bacterial quantity increased significantly in the first three days after inoculation but slowed down afterwards. The new qPCR assay for P. cannabina pv. alisalensis detection will facilitate early detection and disease diagnosis, assist research to provide epidemiological insights for the pathogen, and guide implementation of strategies to manage disease and prevent its spread.

A diazeniumdiolate signal in Pseudomonas syringae upregulates virulence factors and promotes survival in plants.

Pseudomonas syringae infects a wide variety of crops. The mangotoxin-generating operon (mgo) is conserved across many P. syringae strains and is responsible for producing an extracellular chemical signal, leudiazen. Disruption of the mgoA gene in P. syringae pv. syringae (Pss) UMAF0158 alleviated tomato chlorosis caused by this bacterium. We showed that deletion of entire mgo reduced Pss UMAF0158 population in tomato leaflets. Leudiazen restored the signaling activity of the deletion mutant at a concentration as low as 10 nM. Both the diazeniumdiolate and isobutyl groups of leudiazen are critical for this potent signaling activity. Transcriptional analysis showed that mgo and leudiazen induce the expression of mangotoxin biosynthetic operon as well as an uncharacterized gene cluster, RS17235-RS17245. We found that this cluster enhances survival of Pss UMAF0158 in planta and is widely distributed in P. syringae strains. Our results demonstrate that mgo plays prominent roles in the virulence and growth of P. syringae. The mgo and mgo-like signaling systems in different bacteria likely regulate diverse microbe-host interactions.

From lichens to crops: Pathogenic potential of Pseudomonas syringae from Peltigera lichens is similar to worldwide epidemic strains

AbstractThe presence of bacteria belonging to the Pseudomonas syringae complex in the natural vegetation of several Icelandic habitat types has been recently reported, raising questions about the risk to Icelandic crops, particularly given the expected increase in agricultural activity due to climate warming. This study takes advantage of Iceland's unique characteristics and the discovery of P. syringae in Peltigera lichens to gain a better understanding of the potential risk posed by this newly discovered ecological niche. The main objective was to evaluate the pathogenic potential and fitness in crops of P. syringae strains isolated from Peltigera lichen sampled in Iceland, focusing on strains that belong to phylogroups 1 and 2, which commonly contain epidemic strains. The results indicate that P. syringae strains isolated from Icelandic Peltigera lichen have a comparable fitness to epidemic strains in 8 out of 10 tested plant species (rice, tomato, thale cress, annual mugwort, spinach, garlic chives, tobacco and kale). Furthermore, pathogenicity assessment on three plant species highlighted that certain strains also caused similar symptoms and disease severity compared to epidemic strains. These findings provide valuable insights into the potential risks posed by P. syringae from Icelandic natural habitats and illustrate how strains from these habitats have a wide pathogenic potential to crops without having encountered these crops in the last several thousand years of their presence in Iceland.

PYOCIN S5 ACTIVITY AGAINST PSEUDOMONAS SYRINGAE PHYTOPATHOGENIC STRAINS

Pseudomonas syringae is the most frequently isolated bacterial plant pathogens. Previously we revealed a number of Pseudomonas aeruginosa strains – producers of bacteriocins (pyocins) with high activity against P. syringae. However, it was unknown, which type of pyocins possesses the activity against P. syringae strains. S5 and S1 pyocin genes were observed in the genome of most active strain–producer P. aeruginosa UCM В-333. Pyocins were induced by nalidixic acid (100 μg/ml). Using DEAE-cellulose chromatography we isolated two bacteriocin groups from initial lysate. Substances from the 10th fraction had nuclease activity (pyocin S1) and the 27th fraction - pore-forming activity (pyocin S5). It was revealed that S1 pyocin didn’t lead to growth inhibition of studied bacterial plant pathogens. However pyocin S5 caused appearance of growth inhibition area with diameter 14-20 mm. So, pyocins of S1 and S5 types are active substances in P. aeruginosa UCM В-333 lysates, but only S5 pyocin possess activity against P. syringae strains. fpt A receptor gene was revealed in genomes of all analyzed cultures, including used phytopathogenic strains. This fact can explain wide spectrum of pyocin S5 activity. It is suggested that pyocin S5 can be a potential biocontrol agent against phytopathogenic P. syringae strains.

Genetic dissection of the tissue-specific roles of type III effectors and phytotoxins in the pathogenicity of Pseudomonas syringae pv. syringae to cherry.

When compared with other phylogroups (PGs) of the Pseudomonas syringae species complex, P. syringae pv. syringae (Pss) strains within PG2 have a reduced repertoire of type III effectors (T3Es) but produce several phytotoxins. Effectors within the cherry pathogen Pss 9644 were grouped based on their frequency in strains from Prunus as the conserved effector locus (CEL) common to most P. syringae pathogens; a core of effectors common to PG2; a set of PRUNUS effectors common to cherry pathogens; and a FLEXIBLE set of T3Es. Pss 9644 also contains gene clusters for biosynthesis of toxins syringomycin, syringopeptin and syringolin A. After confirmation of virulence gene expression, mutants with a sequential series of T3E and toxin deletions were pathogenicity tested on wood, leaves and fruits of sweet cherry (Prunus avium) and leaves of ornamental cherry (Prunus incisa). The toxins had a key role in disease development in fruits but were less important in leaves and wood. An effectorless mutant retained some pathogenicity to fruit but not wood or leaves. Striking redundancy was observed amongst effector groups. The CEL effectors have important roles during the early stages of leaf infection and possibly acted synergistically with toxins in all tissues. Deletion of separate groups of T3Es had more effect in P. incisa than in P. avium. Mixed inocula were used to complement the toxin mutations in trans and indicated that strain mixtures may be important in the field. Our results highlight the niche-specific role of toxins in P. avium tissues and the complexity of effector redundancy in the pathogen Pss 9644.

Decomposition of dynamic transcriptomic responses during effector-triggered immunity reveals conserved responses in two distinct plant cell populations

Rapid plant immune responses in the appropriate cells are needed for effective defense against pathogens. Although transcriptome analysis is often used to describe overall immune responses, collection of transcriptome data with sufficient resolution in both space and time is challenging. We reanalyzed public Arabidopsis time-course transcriptome data obtained after low-dose inoculation with a Pseudomonas syringae strain expressing the effector AvrRpt2, which induces effector-triggered immunity in Arabidopsis. Double-peak time-course patterns are prevalent among thousands of upregulated genes. We implemented a multi-compartment modeling approach to decompose the double-peak pattern into two single-peak patterns for each gene. The decomposed peaks reveal an “echoing” pattern: the peak times of the first and second peaks correlate well across most upregulated genes. We demonstrated that the two peaks likely represent responses of two distinct cell populations that respond either cell autonomously or indirectly to AvrRpt2. Thus, the peak decomposition has extracted spatial information from the time-course data. The echoing pattern also indicates a conserved transcriptome response with different initiation times between the two cell populations despite different elicitor types. A gene set highly overlapping with the conserved gene set is also upregulated with similar kinetics during pattern-triggered immunity. Activation of a WRKY network via different entry-point WRKYs can explain the similar but not identical transcriptome responses elicited by different elicitor types. We discuss potential benefits of the properties of the WRKY activation network as an immune signaling network in light of pressure from rapidly evolving pathogens.

An atypical 3-ketoacyl ACP synthase III required for acyl homoserine lactone synthesis in Pseudomonas syringae pv. syringae B728a.

The last step of the initiation phase of fatty acid biosynthesis in most bacteria is catalyzed by the 3-ketoacyl-acyl carrier protein (ACP) synthase III (FabH). Pseudomonas syringae pv. syringae strain B728a encodes two FabH homologs, Psyr_3467 and Psyr_3830, which we designated PssFabH1 and PssFabH2, respectively. Here, we explored the roles of these two 3-ketoacyl-ACP synthase (KAS) III proteins. We found that PssFabH1 is similar to the Escherichia coli FabH in using acetyl-acetyl-coenzyme A (CoA ) as a substrate in vitro, whereas PssFabH2 uses acyl-CoAs (C4-C10) or acyl-ACPs (C6-C10). Mutant analysis showed that neither KAS III protein is essential for the de novo fatty acid synthesis and cell growth. Loss of PssFabH1 reduced the production of an acyl homoserine lactone (AHL) quorum-sensing signal, and this production was partially restored by overexpressing FabH homologs from other bacteria. AHL production was also restored by inhibiting fatty acid elongation and providing exogenous butyric acid. Deletion of PssFabH1 supports the redirection of acyl-ACP toward biosurfactant synthesis, which in turn enhances swarming motility. Our study revealed that PssFabH1 is an atypical KAS III protein that represents a new KAS III clade that functions in providing a critical fatty acid precursor, butyryl-ACP, for AHL synthesis.IMPORTANCEAcyl homoserine lactones (AHLs) are important quorum-sensing compounds in Gram-negative bacteria. Although their formation requires acylated acyl carrier proteins (ACPs), how the acylated intermediate is shunted from cellular fatty acid synthesis to AHL synthesis is not known. Here, we provide in vivo evidence that Pseudomonas syringae strain B728a uses the enzyme PssFabH1 to provide the critical fatty acid precursor butyryl-ACP for AHL synthesis. Loss of PssFabH1 reduces the diversion of butyryl-ACP to AHL, enabling the accumulation of acyl-ACP for synthesis of biosurfactants that contribute to bacterial swarming motility. We report that PssFabH1 and PssFabH2 each encode a 3-ketoacyl-acyl carrier protein synthase (KAS) III in P. syringae B728a. Whereas PssFabH2 is able to function in redirecting intermediates from β-oxidation to fatty acid synthesis, PssFabH1 is an atypical KAS III protein that represents a new KAS III clade based on its sequence, non-involvement in cell growth, and novel role in AHL synthesis.

Occurrence of Pseudomonas syringae pvs. actinidiae, actinidifoliorum and Other P. syringae Strains on Kiwifruit in Northern Spain.

Pseudomonas syringae pv. actinidiae (Psa), the agent causing bacterial canker of kiwifruit, has been present in the Principality of Asturias (PA), Northern Spain, since 2013, although with restricted distribution. In this study, 53 strains collected in kiwifruit orchards in PA during the period 2014-2020 were characterized by a polyphasic approach including biochemical and phylogenetic analysis. Thirty-three strains, previously identified by PCR as Psa, have been found to be a homogeneous group in phylogenetic analysis, which seems to indicate that there have been few introductions of the pathogen into the region. Two strains were confirmed as P. syringae pv. actinidifoliorum (Pfm), so this is the first report of Pfm in the PA. The remaining 18 strains were found to be close to P. avellanae and P. syringae pv. antirrhini or to strains described as Pfm look-alikes. Pathogenicity tests carried out on peppers with a selection of strains have shown that both Psa and Pfm caused clear damage, while the 18 atypical strains caused variable lesions. It would be necessary to carry out pathogenicity testing of atypical strains on kiwifruit plants to study the role of these strains in the kiwifruit pathosystem to evaluate their pathogenic potential in this crop.

First Report of Bacterial Leaf Spot on Muskmelon Caused by Pseudomonas syringae pv. syringae in China.

Muskmelon (Cucumis melo L.) is one of the most widely cultivated and economically important fruit crops in the world. In January 2023, muskmelon leaves of cultivar 'Sheng Gu' were observed with irregularly shaped spots in four nurseries in Wanxiang Village, Pudong District of Shanghai, China. Initial symptoms were irregular soaking on the leaves, which progressed to rotting and necrotic spots. The disease incidence of melon seedlings in different nurseries ranged from 10 to 25%. To isolate and identify the causal agent, the small pieces of lesion tissues (5×5 mm) from symptomatic leaves were sterilized in 75% ethanol for 30 s and rinsed three times with sterile water. Following that, tissues were crushed with sterile glass rod in a sterile 2.0 mL centrifuge tube containing 100 μl of sterile water. The suspension was serially diluted before being spread on Luria-Bertani (LB) medium. After 48 h of incubation at 28°C, the cream-colored bacterial colonies from the 10-4 dilution were tiny and purified by streaking on new LB plates. To confirm the species identity of the bacterial isolates, genomic DNA was extracted from four independent representative colonies from different diseased plants, and several conserved genes were amplified and sequenced, including the 16S rRNA gene with primers 27F/1492R, gyrB gene with primers gyrBFor2/gyrBRev2, and rpoD gene with primers rpoDFor2/rpoDRev2 (Lelliot et al. 1966; Murillo et al. 2011). The results showed that the four colonies were identical. Using BLAST analysis in GenBank, the 16S rDNA (accession no. OQ659765, 1,402 bp), the gyrB (accession no. OQ708618, 911 bp), and rpoD sequences (accession no. OQ708619, 798 bp) showed 99.86-100% homology with 99-100% coveage as the corresponding gene sequences in the P. syringae pv. syringae strain HS191 (accession no. CP006256.1). The bacterial isolate was designated as P. syringae pv. syringae strain PDTG. Phylogenetic tree analysis of 16S rDNA, gyrB and rpoD genes further verified that the bacteria isolate was in close proximity to P. syringae pv. syringae. Additionally, all four isolates were detected in PCR with P. syringae pv. syringae specific primers, PsyF/ PsyR (Borschinger et al. 2016; Guilbaud et al. 2016). Ten two weeks old healthy 'Sheng Gu' muskmelon seedlings were inoculated by spraying with a bacterial suspension of 108 CFU/ml, and ten additional healthy plants treated with sterilized water served as the control. The inoculated plants were maintained at 25°C and 75% relative humidity for 7 days in artificial climate room. Water-soaked rot, similar as those seen in the nurseries, appeared on leaves 7 days after inoculation (dai), while the leaves of control plants remained healthy. The bacteria were re-isolated from rot of inoculated leaves and confirmed as the original pathogen by PCR with the PsyF/ PsyR primers and the 16S rRNA gene sequences. To our knowledge, this is the first report of P. syringae pv. syringae causing bacterial leaf spot on muskmelon in China, and this report expands the host range of P. syringae pv. syringae.

Investigating the biosynthesis and roles of the auxin phenylacetic acid during Pseudomonas syringae-Arabidopsis thaliana pathogenesis.

Several plant-associated microbes synthesize the auxinic plant growth regulator phenylacetic acid (PAA) in culture; however, the role of PAA in plant-pathogen interactions is not well understood. In this study, we investigated the role of PAA during interactions between the phytopathogenic bacterium Pseudomonas syringae strain PtoDC3000 (PtoDC3000) and the model plant host, Arabidopsis thaliana. Previous work demonstrated that indole-3-acetaldehyde dehydrogenase A (AldA) of PtoDC3000 converts indole-3-acetaldehyde (IAAld) to the auxin indole-3-acetic acid (IAA). Here, we further demonstrated the biochemical versatility of AldA by conducting substrate screening and steady-state kinetic analyses, and showed that AldA can use both IAAld and phenylacetaldehyde as substrates to produce IAA and PAA, respectively. Quantification of auxin in infected plant tissue showed that AldA-dependent synthesis of either IAA or PAA by PtoDC3000 does not contribute significantly to the increase in auxin levels in infected A. thaliana leaves. Using available arogenate dehydratase (adt) mutant lines of A. thaliana compromised for PAA synthesis, we observed that a reduction in PAA-Asp and PAA-Glu is correlated with elevated levels of IAA and increased susceptibility. These results provide evidence that PAA/IAA homeostasis in A. thaliana influences the outcome of plant-microbial interactions.

In Vivo Production of dsRNA Using Bacteriophage ϕ6 in Pseudomonas syringae Cit7 Cells.

RNA interference (RNAi), also known as post-transcriptional gene silencing (PTGS), is one of the emerging genetic engineering techniques to effectively silence or inhibit the expression of target genes. This chapter describes a method for in vivo production of dsRNA in non-pathogenic Pseudomonas syringae strains using phage ϕ6 RNA-dependent RNA polymerase, extraction and purification of dsRNA from bacterial solution, and the use of dsRNA to induce silencing of green fluorescent protein (GFP) in transgenic Nicotiana benthamiana.

The genetic architecture of the adaptive potential of Arabidopsis thaliana in response to Pseudomonas syringae strains isolated from south‐west France

AbstractPhytopathogens are a threat for global food production and security. Emergence or re‐emergence of plant pathogens is highly dependent on the environmental conditions affecting pathogen spread and survival. Under climate change, a geographic expansion of pathogen distribution poleward has been observed, potentially resulting in disease outbreaks on crops and wild plants. Therefore, estimating the adaptive potential of plants to novel epidemics and describing the underlying genetic architecture is a primary need to propose agricultural management strategies reducing pathogen outbreaks and to breed novel plant cultivars adapted to pathogens that might spread under climate change. To address this challenge, we inoculated Pseudomonas syringae strains isolated from Arabidopsis thaliana populations from south‐west of France on the highly genetically polymorphic TOU‐A A. thaliana population from north‐east France. While no adaptive potential was identified in response to most P. syringae strains, the TOU‐A population displayed a variable disease response to the JACO‐CL strain belonging to the P. syringae phylogroup 7 (PG7). This strain carried a reduced type III secretion system (T3SS) characteristic of the PG7 as well as flexible genomic traits and potential novel effectors. Genome‐wide association mapping on 192 TOU‐A accessions revealed a polygenic architecture of disease response to JACO‐CL. The main quantitative trait locus (QTL) region encompasses two R genes and the AT5G18310 gene encoding ubiquitin hydrolase, a target of the AvrRpt2 P. syringae effector. Altogether, our results pave the way for a better understanding of the genetic and molecular basis of the adaptive potential in an ecologically relevant A. thaliana–P. syringae pathosystem.

Pseudomonas syringae coffee blight is associated with the horizontal transfer of plasmid-encoded type III effectors.

The emergence of new pathogens is an ongoing threat to human health and agriculture. While zoonotic spillovers received considerable attention, the emergence of crop diseases is less well studied. Here, we identify genomic factors associated with the emergence of Pseudomonas syringae bacterial blight of coffee. Fifty-three P. syringae strains from diseased Brazilian coffee plants were sequenced. Comparative and evolutionary analyses were used to identify loci associated with coffee blight. Growth and symptomology assays were performed to validate the findings. Coffee isolates clustered in three lineages, including primary phylogroups PG3 and PG4, and secondary phylogroup PG11. Genome-wide association study of the primary PG strains identified 37 loci, including five effectors, most of which were encoded on a plasmid unique to the PG3 and PG4 coffee strains. Evolutionary analyses support the emergence of coffee blight in PG4 when the coffee-associated plasmid and associated effectors derived from a divergent plasmid carried by strains associated with other hosts. This plasmid was only recently transferred into PG3. Natural diversity and CRISPR-Cas9 plasmid curing were used to show that strains with the coffee-associated plasmid grow to higher densities and cause more severe disease symptoms in coffee. This work identifies possible evolutionary mechanisms underlying the emergence of a new lineage of coffee pathogens.

Unexpected Diversity of Pseudomonads Associated with Bacterial Leaf Spot of Cucurbits in the Southeastern United States.

Bacterial leaf spot of cucurbits (BLS) is an emerging disease in the southeastern United States that is capable of causing widespread outbreaks under conducive conditions. Historically attributed solely to the bacterium Pseudomonas syringae pv. lachrymans, recent studies have identified additional P. syringae pathovars as causal agents of the disease. To further investigate the identity and diversity of P. syringae strains associated with BLS in the southeastern United States, 47 bacterial isolates were recovered from symptomatic cucurbits from Florida, Alabama, and Georgia. Strains were characterized using the LOPAT testing scheme, fluorescence, and pathogenicity to watermelon and squash seedlings. Thirty-eight fluorescent isolates underwent whole-genome sequencing and were further characterized with 16S rRNA, four gene multilocus sequence analysis (MLSA) phylogeny, and average nucleotide identity analysis. Thirty-four isolates were identified as members of the P. syringae species complex, including P. syringae sensu stricto (12), P. alliivorans (12), P. capsici (nine), and P. viridiflava (one). An additional four isolates were found to belong to the Pseudomonas genus outside of the syringae species complex, though they did not share 95% or greater average nucleotide identity to any validly published species and are believed to belong to three novel Pseudomonas species. These results reveal an unpredicted level of diversity of Pseudomonas strains associated with BLS in the region and show the benefits of whole-genome sequencing for strain identification. Identification of P. capsici, which is capable of causing disease at higher temperatures than P. syringae, as a causal agent of BLS may also affect management strategies in the future.

Pseudomonas syringae isolated in lichens for the first time: Unveiling Peltigera genus as the exclusive host.

Pseudomonas syringae is a bacterial complex that is widespread through a range of environments, typically associated with plants where it can be pathogenic, but also found in non-plant environments such as clouds, precipitation, and surface waters. Understanding its distribution within the environment, and the habitats it occupies, is important for examining its evolution and understanding behaviours. After a recent study found P. syringae living among a range of vascular plant species in Iceland, we questioned whether lichens could harbour P. syringae. Sixteen different species of lichens were sampled all over Iceland, but only one lichen genus, Peltigera, was found to consistently harbour P. syringae. Phylogenetic analyses of P. syringae from 10 sampling points where lichen, tracheophyte, and/or moss were simultaneously collected showed significant differences between sampling points, but not between different plants and lichens from the same point. Furthermore, while there were similarities in the P. syringae population in tracheophytes and Peltigera, the densities in Peltigera thalli were lower than in moss and tracheophyte samples. This discovery suggests P. syringae strains can localize and survive in organisms beyond higher plants, and thus reveals opportunities for studying their influence on P. syringae evolution.

Characterization of Pseudomonas syringae strains associated with soybean bacterial blight and in vitro inhibitory effect of oregano and thyme essential oils

Pseudomonas savastanoi pv. glycinea is the causative agent of bacterial blight in soybean. Usually, the control of phytopathogenic bacteria is carried out using toxic pesticides for the environment, which generate rapid bacterial resistance. For this reason, it is suggested to do crop rotation and use certified seeds. New natural compounds are being studied as an alternative to avoid these diseases, like essential oils (EO) which have demonstrated antimicrobial activity. This work aimed to study the antimicrobial activity of EO from Origanum vulgare and Thymus vulgaris against Pseudomonas syringae isolated from soybean bacterial blight in the southern area of Córdoba province. In this study, 14 isolates from Coronel Moldes and General Cabrera fields were identified as P. syringae Group I of LOPAT. The strains P. syringae M3-C1 and P. syringae M7-C1 were identified at the species level by housekeeping genes. EO were obtained by hydro-distillation from thyme and oregano plants. The minimum inhibitory concentration (MIC) was carried out by the broth microdilution technique, determining that both EO could inhibit the isolated soybean strains presenting variations in the active concentrations. The bactericidal activity was also assayed (MBC). These EO were also able to inhibit the biofilm formation of the isolates at sub-inhibitory concentrations. These preliminary studies demonstrated the antimicrobial activity of these EO and their potential use as biopesticides; therefore, future studies will be carried out to develop formulations of these compounds.

Identification and characterization of Pseudomonas syringae pv. syringae strains from various plants and geographical regions

Identification and characterization of Pseudomonas syringae pv. syringae strains from various plants and geographical regions

Characterization of causal agents of bacterial canker on apricot plantations and risk mapping using GIS in Aras Basin (Türkiye)

AbstractBacterial canker of stone fruits caused by Pseudomonas syringae pv. syringae (Pss) and Pseudomonas syringae pv. morsprunorum (Psm race‐1/Psm race‐2) may lead to significant yield and crop losses in apricot (Prunus armeniaca L.) cultivation areas in Türkiye. Strains pathogenic to apricot were isolated from trees with symptoms (mainly necrotized buds and dieback) of bacterial canker in orchards in Aras Basin. Pathogens were characterized using pathogenicity tests, phenotypic assays, end‐point PCR and multilocus sequence analysis (MLSA). Fifteen Pseudomonas syringae strains were isolated from 205 plant samples collected from apricot orchards showing symptoms of bacterial canker. As a consequence of the diagnostic tests, all isolates were identified as P. syringae pv. syringae. In this study, Pss, Psm R1 and Psm R2 strains in stone fruits were separated into different phylogroups (Pg‐2, Pg‐3, sPg‐1b) based on MLSA. Turkish strains obtained from stone fruits, particularly apricot, showed genetic heterogeneity, and clustered in different sub‐phylogroups (sPg‐2b, sPg‐2c, sPg‐2d). All these strains except strain K258 are also clustered in the same sub‐phylogroups (sPg‐2b and sPg‐2d) with other strains from different countries especially Iran, Lebanon, etc. Strain K258 isolated from apricot was clustered in sPg‐2c with Pss strain 642 (USA). The risk of bacterial canker disease in apricot growing areas is considered using GIS in this study. It was determined that a significant part of the Iğdır Plain, the biggest agricultural area in the Aras Basin, is at very high risk.

Phytopathogenic Pseudomonas syringae as a Threat to Agriculture: Perspectives of a Promising Biological Control Using Bacteriophages and Microorganisms

Pseudomonas syringae is a Gram-negative bacterium that infects a wide range of plants, causing significant economic losses in agricultural production. The pathogen exhibits a high degree of genetic and phenotypic diversity, which has led to the classification of P. syringae strains into different pathovars based on their host range and disease symptoms. Copper-based products have traditionally been used to manage infections in agriculture, but the emergence of copper-resistant strains has become a significant concern. Biological control is a promising strategy to manage P. syringae, as it offers an environmentally friendly and sustainable approach to disease management. The review includes an overview of the biology and epidemiology of P. syringae, and of the mechanisms of action of various biological control agents, mainly microorganisms (antagonistic bacteria, and fungi) and bacteriophages. Specifically, this review highlights the renewed interest in bacteriophages (bacteria-infecting viruses) due to their advantages over other eco-friendly management methods, thanks to their bactericidal properties and potential to target specific pathogenic bacteria. The potential benefits and limitations of biological control are also examined, along with research directions to optimize the use of this approach for the management of P. syringae.

VIRULENCE OF PSEUDOMONAS SYRINGAE STRAINS FROM SORIZ (SORGHUM ORYSOIDUM) AND THEIR SPECIALIZATION

Objective. To study the virulence properties of Pseudomonas syringae strains isolated from soriz and various representatives of sorghum crops in Ukraine and to establish the sensitivity range of plants to these pathogens. Methods. Ten strains freshly isolated from soriz (Sorghum orysoidum) were used to study virulence properties, namely: Pseudomonas syringae 210342, 211141a, 211141, 210341, 21034, 210541b, 210541, and Pseudomonas sp. 210521, 21052, 21054, pathogenic strains of sorghum crops from the collection of the Department of Phytopathogenic Bacteria of D. K. Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine and collection strains of phytopathogens that can be potential causative agents of bacterial diseases of soriz or are causative agents of diseases of agricultural crops that can be used in crop rotation. The virulence properties of strains isolated from sorghum were determined on plants of the genus Sorghum. To establish the range of crops sensitive to these pathogens, a number of plants belonging to other genera and families, both cultivated and weed, were tested. Artificial inoculation was carried out in the greenhouse and in the field as injection of bacterial suspension of 1×109 CFU/mL into the leaf surface and stem at all phases of the development of soriz and sorghum cultures. Accounting for artificial contamination was carried out in 7 to 14 days using a locally modified 6-point scale. Aggressiveness of bacteria was assessed by the size of necrosis that formed at the site of injection of the bacterial suspension. Aggressiveness was calculated as the arithmetic mean of the results. Results. P. syringae strains isolated from soriz were found to be virulent for soriz and sorghum crops, however, heterogeneous in aggressiveness. The most aggressive strains were P. syringae 211141a, 210341, and 21034. Some collection strains, isolated from different cultures, showed higher aggressiveness against sugar and grain sorghum than against soriz and Sudan grass. The studied strains of phytopathogens were found to affect segetal vegetation; the most sensitive to the studied strains were horsetail, field bindweed, goldenrod. They showed high aggressiveness against setaria, ragweed, and common sedge. Strains of P. syringae affecting potato, beet, sunflower and tomato plants were identified. Conclusion. Pseudomonas syringae strains isolated from contaminated sorghum plants are virulent and capable of infecting a wide range of plants, including plants of the sorghum family, other types of agricultural crops, and various types of segetal vegetation. Strains of Pectobacterium carotovorum, Xanthomonas vasicola pv. holcicola, Diskeya chryzanthemy, Pantoea agglomerans can be potential pathogens of sorghum crops. The results indicate that segetal vegetation can serve as a reservoir of bacterial infections and play a role in the transfer of bacterial pathogens.