Agent Of Blight Research Articles

Performance of Novel Antimicrobial Protein Bg_9562 and In Silico Predictions on Its Properties with Reference to Its Antimicrobial Efficiency against Rhizoctonia solani.

Bg_9562 is a potential broad-spectrum antifungal effector protein derived from the bacteria Burkholderia gladioli strain NGJ1 and is effective against Rhizoctonia solani, the causal agent of sheath blight in rice. In the present study, in vitro antifungal assays showed that Bg_9562 was efficient at 35 °C and 45 °C and ineffective either at high acidic pH (3.0) or alkaline pH (9.5) conditions. Compatibility studies between the native bioagents Trichoderma asperellum TAIK1 and Bacillus subtilis BIK3 indicated that Bg_9562 was compatible with the bioagents. A field study using foliar spray of the Bg_9562 protein indicated the need of formulating the protein before its application. In silico analysis predicted that Bg_9562 possess 111 amino acid residues (46 hydrophobic residues, 12 positive and 8 negative residues) with the high aliphatic index of 89.92, attributing to its thermostability with a half-life of 30 h. Bg_9562 (C491H813N137O166S5) possessed a protein binding potential of 1.27 kcal/mol with a better possibility of interacting and perturbing the membrane, the main target for antimicrobial proteins. The secondary structure revealed the predominance of random coils in its structure, and the best 3D model of Bg_9562 was predicted using an ab initio method with Robetta and AlphaFold 2. The predicted binding ligands were nucleic acids and zinc with confidence scores of 0.07 and 0.05, respectively. The N-terminal region (1–14 residues) and C-terminal region (101 to 111) of Bg_9562 residues were predicted to be disordered regions. Stability and binding properties of the protein from the above studies would help to encapsulate Bg_9562 using a suitable carrier to maintain efficiency and improve delivery against Rhizoctonia solani in the most challenging rice ecosphere.

The periplasmic chaperone protein Psg_2795 contributes to the virulence of Pseudomonas savastanoi pv. glycinea: the causal agent of bacterial blight of soybean.

Pseudomonas savastanoi pv. glycinea (Psg also named P. syringae pv. glycinea and P. amygdali pv. glycinea) is the causative agent of bacterial blight in soybean. The identification of virulence factors is essential for understanding the pathogenesis of Psg. In this study, a mini-Tn5 transposon mutant library of Psg strain PsgNC12 was screened on soybean, and one low-virulent mini-Tn5 mutant, designated as 4573, was identified. Sequence analysis of the 4573-mutant revealed that the mini-Tn5 transposon was inserted in the Psg_2795 gene. Psg_2795 encodes a FimC-domain protein that is highly conserved in Pseudomonas. Further analysis revealed that the mutation and knockout of Psg_2795 results in a reduced virulence phenotype on soybean, decreased motility, weakened bacterial attachment to a glass surface and delayed the population growth within soybean leaves. The phenotype of the 4573-mutant could be complemented nearly to wild-type levels using an intact Psg_2795 gene. Collectively, our results demonstrate that Psg_2795 plays an important role in the virulence, motility, attachment and the population growth of PsgNC12 in soybean. This finding provides a new insight into the function of periplasmic chaperone proteins in a type I pilus and provides reference information for identifying Psg_2795 homologues in P. savastanoi and other bacteria.

Comparison of the Antagonistic Potential of the Entomopathogenic Bacterium Serratia nematodiphila GCSR38 with other Effective Microorganisms for the Control of Rice Bacterial Leaf Blight

The efficacy of antagonistic microorganisms and secondary metabolites of entomopathogenic bacteria was evaluated in vitro and in vivo against Xanthomonas oryzae pv. oryzae (Xoo), a causal agent of rice bacterial leaf blight. The effect of the culture filtrates of the microorganisms was tested against Xoo, using the agar well diffusion assay. Trichoderma sp. UB05/3 was the most effective against Xoo. However, the results were not statistically different from those of the entomopathogenic bacterium Serratia nematodiphila GCSR38 with a 22.97 mm and 19.15 mm zone of inhibition, respectively. The effect of the secondary metabolite crude extracts of antagonistic microorganisms (2,000 µg/ml) on Xoo inhibition was tested by the paper disc diffusion method. Only S. nematodiphila GCSR38 was able to control Xoo, with an inhibition zone of 17.60 mm. Minimum inhibitory concentration (MIC) was determined using a 96-well microtiter plate. The MIC of secondary metabolites crude extracts of S. nematodiphila GCSR38 was 1,000 µg/ml and the minimum bactericidal concentration (MBC) was 1,000 µg/ml. The efficacy of the secondary metabolite crude extract of S. nematodiphila GCSR38 against Xoo was tested using the detached leaf technique; the secondary metabolite crude extracts controlled the disease, with 24.45% and 15.56% disease severity when used before and after inoculation with Xoo, respectively. Inoculation with Xoo alone resulted in a disease severity of 44.45%. The secondary metabolite crude extracts of S. nematodiphila GCSR38 can reduce disease severity of bacterial leaf blight in rice by 28.89%, whereas zinc thiazole causes a disease severity of 22.22% and Xoo alone causes a disease severity of 66.67%.

Alternaria spp. Associated with Leaf Blight of Maize in Heilongjiang Province, China.

Maize (Zea mays L.) is a major economic crop worldwide. Maize can be infected by Alternaria species causing leaf blight that can result in significant economic losses. In this study, 168 Alternaria isolates recovered from symptomatic maize leaves were identified based on morphological characteristics, pathogenicity, and multilocus sequence analyses of the genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), the internal transcribed spacer of ribosomal DNA (rDNA ITS), the RNA polymerase II second largest subunit (RPB2), and histone3 (HIS3). Maize isolates grouped to four Alternaria species including Alternaria tenuissima, A. alternata, A. burnsii, and Alternaria sp. Notably, A. tenuissima (71.4%) was the most prevalent of the four isolated species, followed by A. alternata (21.5%), Alternaria sp. (4.1%), and A. burnsii (3.0%). Pathogenicity tests showed that all four Alternaria species could produce elliptic to nearly round, or strip, lesions on leaves of maize, gray-white to dry white in the lesion centers and reddish-brown at the edges. The average disease incidence (58.47%) and average disease index (63.55) of maize leaves inoculated with A. alternata were significantly higher than levels resulting from A. tenuissima (55.28% and 58.49), Alternaria sp. (55.34% and 58.75), and A. burnsii (56% and 55). Haplotype analyses indicated that there were 14 haplotypes of A. tenuissima and five haplotypes of A. alternata in Heilongjiang Province and suggested the occurrence of a population expansion. Results of the study showed that Alternaria species associated with maize leaf blight in Heilongjiang Province are more diverse than those that have been previously reported. This is the first report globally of A. tenuissima, A. burnsii, and an unclassified Alternaria species as causal agents of leaf blight on maize.

Genome Resource of a Hypervirulent Strain C9-3 of Xanthomonas oryzae pv. oryzae Causing Bacterial Blight of Rice

Xanthomonas oryzae pv. oryzae is the causal agent of bacterial blight, one of the most devastating diseases of rice. Here, a hypervirulent strain, C9-3, defeating Xa1, Xa10, xa13, and Xa23 resistance genes, was used to extract genomic DNA for single molecule real-time (SMRT) sequencing. After assembly, the genome consists of a single-circular chromosome with the size of 4,924,298 bp with G+C content of 63.7% and contains 4,715 genes. Annotation and analysis of the TALE genes using a suite of applications named AnnoTALE suggested that 17 transcription activator-like effectors, including 15 typical TALEs and 2 iTALEs/truncTALEs, were encoded in the genome. The approach and genome resource will contribute to the discovery of new virulence effectors and understanding on rice-X. oryzae pv. oryzae interactions.

Response of Transgenic Potato Plants Expressing Heterologous Genes of ∆9- or ∆12-Acyl-lipid Desaturases to Phytophthora infestans Infection.

Late blight is one of the most economically important diseases affecting potato and causing a significant loss in yield. The development of transgenic potato plants with enhanced resistance to infection by Phytophthora infestans may represent a possible approach to solving this issue. A comparative study of the leaf response in control potato plants (S.tuberosum L. cultivar Skoroplodnyi), control transgenic plants expressing the reporter gene of thermostable lichenase (transgenic licBM3 line) and transgenic plants expressing cyanobacterial hybrid genes ∆9-acyl-lipid desaturase (transgenic desC lines) and ∆12-acyl-lipid desaturase (transgenic desA lines) to infection with P. infestans has been performed. The expression of desaturase genes in potato plants enhanced their tolerance to potato late blight agents as compared with the control. The lipid peroxidation level raised in the leaves of the control and transgenic desA plants on third day after inoculation with P. infestans zoospores and remained the same in the transgenic desC plants. The number of total phenolic compounds was increased as early as on the second day after infection in all studied variants and continued to remain the same, except for transgenic desC plants. Accumulation of flavonoids, the main components of the potato leaf phenolic complex, raised on the second day in all studied variants, remained unchanged on the third day in the control plants and decreased in most transgenic plants expressing desaturase genes. The results obtained in our study demonstrate that the expression of genes of Δ9- and Δ12-acyl-lipid desaturases in potato plants enhanced their resistance to P. infestans as compared with the control non-transgenic plants due to concomitant accumulation of phenolic compounds, including flavonoids, in the leaves. All these changes were more pronounced in transgenic desC plants, which indicates that the Δ9-acyllipid desaturase gene appears to be a potential inducer of the production of biological antioxidants in plant cells.

Nigrospora sphaerica Causing Leaf Blight Disease on Sesame in Pakistan

Sesame (Sesamum indicum L.) commonly known as 'til' is the most ancient and widely grown oilseed crop of Pakistan. During 2020, field survey conducted in various research fields of Faisalabad (31°26'00.2″N, 73°04'25.4″E) revealed the occurrence of characteristic leaf blight disease with an incidence of 10 to 13%. The symptoms were characterized by yellow-brown and irregular lesions. At later stages, the lesions expanded and the affected leaves turned grayish to dark-brown and finally became wilted. Symptomatic leaves (both the diseased and healthy tissue) were cut into approximately 2 × 2 mm pieces, surface sterilized with 1% sodium hypochlorite for 30 s, 70% ethanol for 30 s, and finally, three times in sterile distilled water prior to culturing on Potato Dextrose Agar (PDA) and incubated at 25 °C under a 12-h photoperiod for 7 days. To obtain pure cultures, hyphal tips of growing mycelia from leaf tissues were carefully isolated and transferred onto fresh PDA plates. Fungal colonies of 11 isolates were initially white, becoming light to dark-gray. The conidia were black, spherical to subspherical, and single-celled (12 to 14 × 18 to 20 μm) in diameter, which were borne on a hyaline vesicle at the tip of the conidiophore. Further, to identify the pathogen to the species level, genomic DNA was extracted using a modified CTAB protocol described by (Guo et al. 2000). The internal transcribed spacer (ITS) region of the ribosomal DNA and translation elongation factor 1-alpha (TEF1-α) gene were amplified using ITS1/ITS4 (White et al. 1990) and EF1-728F/EF1-986R primer sets (Carbone and Kohn 1999), respectively. The sequences were submitted to GenBank (accession no. MW287214 for ITS and MW325222 for TEF1. The sequences comparison revealed 99% and 100% similarity to multiple sequences of N. sphaerica (GenBank accessions KX834822 and MN995332). On the basis of cultural features, conidial morphology and molecular data, the fungus was identified as Nigrospora sphaerica (Sacc.) Mason (Wang et al. 2017; Chen et al. 2018). To test the pathogenicity, fresh and healthy leaves of ten 6-week-old growth stage sesame plants were spray inoculated with a conidial suspension (105 conidia/ml), collected from a 7-day-old culture on PDA. In addition, 10 plants sprayed with sterile distilled water served as controls. Inoculated plants were covered with polyethylene bags to maintain high humidity and kept at 28°C, and observations were made at regular intervals. After 8-10 days of inoculation, leaves developed blight symptoms similar to those observed on naturally infected leaves, whereas control leaves remained asymptomatic. The pathogen was re-isolated from the inoculated leaves, and its identity was confirmed by morphological and molecular (ITS and TEF1) means, thus fulfilling Koch's postulates. N. sphaerica is distributed on a wide range of hosts and has been reported from different host genera including monocotyledonous and dicotyledonous (Wang et al. 2017). Previously, N. sphaerica has been reported to cause leaf blight of Cunninghamia lanceolata in China (Xu and Liu, 2016). To the best of our knowledge, this is the first report of Nigrospora sphaerica as the causal agent of leaf blight of sesame in Pakistan. Because sesame is an important oilseed crop of Pakistan, appropriate disease management practices should be developed and implemented.

Microsatellite Genotyping in the Chestnut Blight Fungus Cryphonectria parasitica.

This chapter describes the use of polymorphic microsatellite (simple sequence repeats, SSR) markers for genotyping isolates of Cryphonectria parasitica, the causal agent of chestnut blight. The SSR presented are particularly useful to characterize the genetic population structure of this invasive fungal pathogen, including invasion history (e.g., possible source population, introduced genotypes) and reproduction mode (sexual vs. asexual). Microsatellite markers can also be used to track fungal strains in laboratory and field experiments.

Comparative proteome profiling of susceptible and resistant rice cultivars identified an arginase involved in rice defense against Xanthomonas oryzae pv. oryzae

Xanthomonas oryzae pv. oryzae (Xoo), the causative agent of bacterial blight, is one of the major threats to rice productivity. Yet, the molecular mechanism of rice-Xoo interaction is elusive. Here, we report comparative proteome profiles of Xoo susceptible (Dongjin) and resistant (Hwayeong) cultivars of rice in response to two-time points (3 and 6 days) of Xoo infection. Low-abundance proteins were enriched using a protamine sulfate (PS) precipitation method and isolated proteins were quantified by a label-free quantitative analysis, leading to the identification of 3846 proteins. Of these, 1128 proteins were significantly changed between mock and Xoo infected plants of Dongjin and Hwayeong cultivars. Based on the abundance pattern and functions of the identified proteins, a total of 23 candidate proteins were shortlisted that potentially participate in plant defense against Xoo in the resistant cultivar. Of these candidate proteins, a mitochondrial arginase-1 showed Hwayeong specific abundance and was significantly accumulated following Xoo inoculation. Overexpression of arginase 1 (OsArg 1) in susceptible rice cultivar (Dongjin) resulted in enhanced tolerance against Xoo as compared to the wild-type. In addition, expression analysis of defense-related genes encoding PR1, glucanase I, and chitinase II by qRT-PCR showed their enhanced expression in the overexpression lines as compared to wild-type. Taken together, our results uncover the proteome changes in the rice cultivars and highlight the functions of OsARG1 in plant defense against Xoo.

Aqueous extracts from Acalypha gaumeri and Bonellia flammea for leaf blight control in chrysanthemum (Chrysanthemum morifolium)

The chrysanthemum is the second most important cut flower in the world, however, its quality and commercial value is affected by the leaf blight produced by <em>Alternaria </em>spp. The objective of this work was to evaluate the causal agent of leaf blight in Chrysanthemum, and its control with aqueous extracts of <em>Acalypha gaumeri </em>and <em>Bonellia flammea</em>. The fungus was collected and identified from leaves and stems of chrysanthemum plants. Subsequently, molecular identification and pathogenicity tests were performed on chrysanthemum plants. In the field, treatments were evaluated with weekly applications of: T1: <em>B. flammea </em>bark extract, T2: <em>A. gaumeri </em>root extract, T3: negative control (water) and T4: Captan® fungicide. Prior to the application of the treatments, plants were inoculated with the isolated fungus (2.5 × 106 spores mL-1) and severity was evaluated. <em>Alternaria chrysanthemi </em>was identified as the causal agent. Based on the severity percentage, the lowest averages of the area under the disease progress curve, the lowest rates of apparent infection, the lowest intensity of the disease and the greater effectiveness in controlling the disease were observed for T2 (165, 0.017, 8 and 67%, respectively) followed by T1 (186, 0.022, 13 y 50 %, respectively) and T4 (179, 0.023, 14 y 45%, respectively), observing a significantly different than negative control T3 (369, 0.025, 25 and 0%, respectively). Plant extracts have potential to be used as an alternative in the management of <em>Alternaria </em>leaf blight in chrysanthemum.

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

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

Comparative Study of Four Coloured Nanoparticle Labels in Lateral Flow Immunoassay.

The detection limit of lateral flow immunoassay (LFIA) is largely determined by the properties of the label used. We compared four nanoparticle labels differing in their chemical composition and colour: (1) gold nanoparticles (Au NPs), red; (2) Au-core/Pt-shell nanoparticles (Au@Pt NPs), black; (3) latex nanoparticles (LPs), green; and (4) magnetic nanoparticles (MPs), brown. The comparison was carried out using one target analyte—Erwinia amylovora, the causal bacterial agent of fire blight. All nanoparticles were conjugated with antibodies through methods that provide maximum functional coverage like physical adsorption (Au NPs, Au@Pt NPs) and covalent bonding (LPs, MPs). All conjugates demonstrated the same ability to bind with E. amylovora through enzyme-linked immunosorbent assay where optical properties of the nanoparticles do not determine the registered signal. However, half-maximal binding was achieved at different numbers of nanoparticles because they differ in size. All conjugates based on four nanoparticle labels were used for lateral flow assays. As a result, Au@Pt NPs provided the minimal detection limit that corresponded to 103 CFU/mL. Au NPs and LPs detected 104 CFU/mL, and MPs detected 105 CFU/mL. The results highlight that simply choosing a coloured label can significantly affect the detection limit of LFIA.

Evaluation of Chestnut Susceptibility to Cryphonectria parasitica: Screening under Controlled Conditions

Cryphonectria parasitica (Murrill) M.E. Barr (Sordariomycetes, Valsaceae) is the causal agent of chestnut blight. This disease is a major concern for chestnut cultivation in Europe. The fungus colonizes vascular tissues and evolves generating cankers causing severe dieback and the death of the tree. Excised and debarked well-lignified shoots of 28 C. sativa genotypes (assay A) and of 10 progenies (assay B) were inoculated with C. parasitica strain FMT3bc2 (vcg: EU2). Fungal growth was measured along the longitudinal axis on the 3rd and 6th days after inoculation. Results indicated the inoculation methodology works and the results were clear after 6 days. Differences in susceptibility to chestnut blight among C. sativa trees of Montseny have been detected both at the individual genotype level and at the progeny level. Nineteen genotypes and four progenies showed a susceptibility to Blight not significantly different from C. mollissima. The methodology was easy to apply in extensive/preliminary selection screenings to assess the susceptibility of C. sativa materials to the Blight.

Suppressing chlorophyll degradation by silencing OsNYC3 improves rice resistance to Rhizoctonia solani, the causal agent of sheath blight.

SummaryNecrotrophic fungus Rhizoctonia solani Kühn (R. solani) causes serious diseases in many crops worldwide, including rice and maize sheath blight (ShB). Crop resistance to the fungus is a quantitative trait and resistance mechanism remains largely unknown, severely hindering the progress on developing resistant varieties. In this study, we found that resistant variety YSBR1 has apparently stronger ability to suppress the expansion of R. solani than susceptible Lemont in both field and growth chamber conditions. Comparison of transcriptomic profiles shows that the photosynthetic system including chlorophyll biosynthesis is highly suppressed by R. solani in Lemont but weakly in YSBR1. YSBR1 shows higher chlorophyll content than that of Lemont, and inducing chlorophyll degradation by dark treatment significantly reduces its resistance. Furthermore, three rice mutants and one maize mutant that carry impaired chlorophyll biosynthesis all display enhanced susceptibility to R. solani. Overexpression of OsNYC3, a chlorophyll degradation gene apparently induced expression by R. solani infection, significantly enhanced ShB susceptibility in a high‐yield ShB‐susceptible variety ‘9522’. However, silencing its transcription apparently improves ShB resistance without compromising agronomic traits or yield in field tests. Interestingly, altering chlorophyll content does not affect rice resistance to blight and blast diseases, caused by biotrophic and hemi‐biotrophic pathogens, respectively. Our study reveals that chlorophyll plays an important role in ShB resistance and suppressing chlorophyll degradation induced by R. solani infection apparently improves rice ShB resistance. This discovery provides a novel target for developing resistant crop to necrotrophic fungus R. solani.

Evaluating eco-friendly botanicals as alternatives to synthetic fungicides against the causal agent of early blight of Solanum melongena

Evaluating eco-friendly botanicals as alternatives to synthetic fungicides against the causal agent of early blight of Solanum melongena

First report of Erwina amylovora in Tuscany, Italy

2-years-old plants of Pyrus communis showing symptoms of fire blight disease were sampled in an orchard in Tuscany (Italy) during Autumn 2020. Plants were obtained the previous spring from a commercial nursery located in a region where the disease is present since 1994. The collected material was processed in the lab in order to verify the presence of the bacterium Erwinia amylovora, the causal agent of fire blight. Pure isolates showing white mucoid colonies and levan producers on Levan medium were putatively assimilated to E. amylovora. DNA was extracted from the cultures and analysed with three molecular assays, including duplex PCR of the 29-Kb plasmid pEA29 and the ams chromosomal region, sequencing of the 16S rDNA and recA gene regions, two real-time PCR assays on symptomatic plant tissues. All tests confirmed the presence of E. amylovora. Symptomatic and surrounding plants were removed and immediately destroyed according to the regional phytosanitary protocol. This outcome poses a serious threat for fruit orchards in the area.

CLONING AND EXPRESSION OF A MURE LIGASE ENZYME AS A POTENTIAL TARGET AGAINST BACTERIA XANTHOMONAS ORYZAE PV. ORYZAE

Xanthomonas oryzae pv. oryzae (Xoo) is causal agent of bacterial blight (BB) in rice. Many genes in Xoo have been identified in recently years. One of these genes, a gene coded for uridine diphosphate (UDP)-MurNAc-tripeptide ligase (MurE), catalyses the addition of meso-diaminopimelic acid (m-DAP) into peptidoglycan by coupled to the hydrolysis of ATP has more popular interest. However, there are no experimental data to confirm hypothesis of this enzyme in Xoo. A significant overview at the ATP binding site of most the MurE ligases demonstrated much more variable with amino acid sequence identities in this part, variable percentage around 22 to 26%. Besides, a refined homology structural feature between EcMurE and XooMurE will very important for determining possible involvement of the MurE ligase activity in Xoo. Therefore, a new recombinant protein named XooMurE from Xoo was purified with the N-terminal His-tagged form through a Ni-NTA column in this study. After purification, the Histag was removed then out of the N-terminal His-tagged XooMurE by TEV protease. Purification effectiveness of XooMurE over 95% in this study could produce an essential material for e studies about mechanism of XooMurE and consequently available direction for discovering novel anti-bacterial compounds against Xanthomonas oryzae pv. oryzae (xoo).

Determining the tolerance of the Turkish hazelnut cultivars (Coryllus avellana) against the pathovar Xanthomonas arboricola pv. corylina (causing agent of fireblight)

Objective: This study was conducted to define the tolerance of various hazelnut cultivars against Xanthomonas arboricola pv. corylina causing agent of bacterial blight. 
 Material and Methods: The Turkish hazelnut cultivars (Acı, Çakıldak, Foşa, İncekara, Kalınkara, Kan, Kargalak, Kuş, Mincane, Palaz, Sivri, Tombul, Uzunmusa, Yassı Badem and Yuvarlak Badem) were used as plant material. AT8/Z strain of Xanthomonas arboricola pv. corylina isolated from the field was tested. Parameters measured to detect bacterial blight tolerance were infected leaf percentage and leaf lesion rate. 
 Results: İncekara showed the minimum percentage of infected leaves (%17.57) and Yassı Badem reflected the highest (%77.51). Yassı Badem had the highest leaf lesion degree (3.00) and İncekara the lowest (1.34). The results suggested that Yassı Badem cultivar was the most susceptible and İncekara the most resistant to the pathogen. The highest antimicrobial activity was represented by Mincane with leaf diameter of 20 mm.
 Conclusion: In this study, resistance to Xanthomonas arboricola pv. corylina pathogen tolerance of Turkish hezelnut cultivars grown in our country was determined. It is thought that the results obtained from the study will shed light on the breeding studies of resistant varieties against this pathogen in the future.

Epidemiology of Fire Blight in Fruit Crops in Kazakhstan

A survey on 13 regions of Kazakhstan was carried out to identify foci of fire blight, the extent of its spread, the range of affected crops, and the identification of infection reserves. The disease was mainly concentrated in the main fruit growing zone.at the south and southeast of Kazakhstan. Symptoms of fire blight were characteristic of two bacterial diseases: namely, necrosis, by Pseudomonas syringae Van. Hall. and fire blight by Erwinia amylovora (Burrill) Winslow et al. The authors performed bacteriological analyses to isolate and identify the causative agent of fire blight from various fruit and wild cultures of the Rosaceae family, using classical bacteriological and molecular genetic methods. Two types of bacteria were isolated from the samples affected by the disease, namely E. amylovora, the causative agent of fire blight, and P. syringae, the causative agent of bacterial necrosis. The results of the studies on the identification of bacterial species E. amylovora and P. syringae were confirmed by Swiss scientists from the Agroscope research center based on an immuno-chromatographic test and by Russian scientists at the All-Russian Research Institute of Plant Quarantine using enzyme-linked immunosorbent assay, FLASH polymerase chain reaction, and polymerase chain reaction in real-time.

Characterization of the biocontrol activity of three bacterial isolates against the phytopathogen Erwinia amylovora.

Antibiotics are sprayed on apple and pear orchards to control, among other pathogens, the bacterium Erwinia amylovora, the causative agent of fire blight. As with many other pathogens, we observe the emergence of antibiotic‐resistant strains of E. amylovora. Consequently, growers are looking for alternative solutions to combat fire blight. To find alternatives to antibiotics against this pathogen, we have previously isolated three bacterial strains with antagonistic and extracellular activity against E. amylovora, both in vitro and in planta, corresponding to three different bacterial genera: Here, we identified the inhibitory mode of action of each of the three isolates against E. amylovora. Isolate Bacillus amyloliquefaciens subsp. plantarum (now B. velezensis) FL50S produces several secondary metabolites including surfactins, iturins, and fengycins. Specifically, we identified oxydifficidin as the most active against E. amylovora S435. Pseudomonas poae FL10F produces an active extracellular compound against E. amylovora S435 that can be attributed to white‐line‐inducing principle (WLIP), a cyclic lipopeptide belonging to the viscosin subfamily (massetolide E, F, L, or viscosin). Pantoea agglomerans NY60 has a direct cell‐to‐cell antagonistic effect against E. amylovora S435. By screening mutants of this strain generated by random transposon insertion with decreased antagonist activity against strain S435, we identified several defective transposants. Of particular interest was a mutant in a gene coding for a Major Facilitator Superfamily (MFS) transporter corresponding to a transmembrane protein predicted to be involved in the extracytoplasmic localization of griseoluteic acid, an intermediate in the biosynthesis of the broad‐spectrum phenazine antibiotic d‐alanylgriseoluteic acid.