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Soft rot‐inducing bacterial pathogens are considered among the main biotic constraints on vegetables and annual crops. Most of the bacterial soft rot pathogens are members of Pectobacterium spp. and Dickeya, with the former genus encompassing a continuously increasing number of species varying in geographic distribution and host of isolation. In Iran, Pectobacterium species have rarely been investigated using high‐throughput nucleotide sequence‐based techniques, leaving the genetic diversity of these pathogens mostly undetermined. During 2021, vegetables and annual crops with symptoms of soft rot and maceration were collected from Fars, Hamedan, Isfahan, Khuzestan and Kohgiluyeh‐Boyer‐Ahmad provinces of Iran. Forty‐four pectinolytic bacterial strains were isolated from symptomatic tissues of bell pepper, cabbage, carrot, lettuce, potato and zucchini. The bacterial strains were subjected to pathogenicity and host range assays, specific PCR tests and phylogenetic analysis of three housekeeping genes (dnaX, gapA and leuS). All strains were pathogenic on their host of isolation. Among the 44 pectinolytic strains, 26 strains were identified as members of Pectobacterium based on PCR using the primer pair Y1/Y2. Multilocus sequence analysis confirmed the latter bacterial strains belonging to the genus Pectobacterium and revealed that the strains scattered into four species: P. brasiliense (five strains), P. carotovorum (eight strains), P. polaris (five strains) and P. versatile (eight strains). P. polaris and P. versatile have not previously been reported in the country. Host range assays showed that P. brasiliense and P. carotovorum have a broader host range than the other two species. [ABSTRACT FROM AUTHOR]

Bacterial canker and spot of stone fruits and bacterial blight of walnut, caused by different pathovars of Xanthomonas arboricola, namely X. arboricola pv. pruni and X. arboricola pv. juglandis, respectively, are considered among quarantine or regulated (nonquarantine) agents in different countries. The two pathogens were initially reported in Iran in 2005 and 1977, respectively. However, the precise identity, pathogenicity status and geographic distribution of these pathogens have been questioned. In this study, we have employed a series of comprehensive field surveys and samplings over a course of 5 years (2015–2019) across 16 provinces to reassess and clarify the occurrence and distribution of X. arboricola on fruit and nut trees in Iran. Besides dozens of non‐Xanthomonas and nonpathogenic bacterial strains, 27 X. arboricola strains were isolated from stone fruits (n = 13) and walnut (n = 14). Except for the strains Peh35, Peh78 and Peh79, the stone fruit strains were nonpathogenic on their host of isolation as well as several other plant species, while the walnut strains were pathogenic on their host of isolation. Multilocus sequence analysis (MLSA) of four housekeeping genes, atpD, dnaK, gyrB and rpoD, revealed that among the stone fruit strains only Peh78 and Peh79 could be assigned into X. arboricola pv. pruni, while the walnut strains were identified as X. arboricola pv. juglandis. Based on the results of pathogenicity assays and phylogenetic analyses, we confirm the occurrence of X. arboricola pv. juglandis in Iran, while X. arboricola pv. pruni was absent in the plant samples evaluated in this study. [ABSTRACT FROM AUTHOR]

Curtobacterium flaccumfaciens (Microbacteriaceae), a plant-pathogenic coryneform species includes five pathovars with valid names and a number of proposed – but unvalidated – new members. In this study, phenotypic features and DNA similarity indexes were investigated among all C. flaccumfaciens members. Results showed that the C. flaccumfaciens pv. poinsettiae strains causing bacterial canker of Euphorbia pulcherrima in the USA as well as the orange-/red-pigmented strains of C. flaccumfaciens pv. flaccumfaciens pathogenic on dry beans in Iran are too distinct from each other and from the type strain of the species to be considered members of C. flaccumfaciens. Hence, the latter two groups were elevated at the species level as C. poinsettiae sp. nov. (ATCC 9682T = CFBP 2403T = ICMP 2566T = LMG 3715T = NCPPB 854T as type strain), and C. aurantiacum sp. nov. (50RT = CFBP 8819T = ICMP 22071T as type strain). Within the emended species C. flaccumfaciens comb. nov., yellow-pigmented strains causing bacterial wilt of dry beans and those causing bacterial canker of Euphorbia pulcherrima in Europe were retained as C. flaccumfaciens pv. flaccumfaciens and C. flaccumfaciens pv. poinsettiae, respectively; while taxonomic position of the sugar beet pathogen C. flaccumfaciens pv. beticola ATCC BAA144PT was confirmed. The newly described onion pathogen C. allii was also reclassified as C. flaccumfaciens pv. allii with the pathotype strain LMG 32517PT. Furthermore, C. flaccumfaciens pv. basellae causing bacterial leaf spot of malabar spinach (Basella rubra) was transferred to C. citreum pv. basellae with ATCC BAA143PT as pathotype. [ABSTRACT FROM AUTHOR]

During July 2015 to June 2017, 63 walnut and 94 stone fruit orchards were surveyed in Iran to identify and characterize the causal agent of a disease inducing leaf spot and fruit canker symptoms. Altogether, 189 Prunus spp. and 141 walnut samples (i.e. leaves, branches and fruits) were collected from 10 Provinces in northern, northwestern, central and southern Iran. Forty-three yellow-pigmented Gram-negative bacterial isolates were recovered from symptomatic plants. Only seven bacterial strains isolated from walnut in Mazandaran and Golestan Provinces were identified as X. arboricola pv. juglandis based on pathogenicity assays, specific PCRs, and multilocus sequence analysis. Based on the phenotypic features and phylogenetic analysis of gyrB gene sequences, strains isolated from stone fruit trees were identified as multiple Pantoea species (i.e. P. agglomerans, P. ananatis, P. vagans and P. pleuroti). None of these strains caused symptoms on Prunus spp. plantlets tested under greenhouse conditions. Although the occurrence of bacterial blight of walnut in Iran was ascertained using molecular-phylogenetic analysis, no evidence was obtained that the observed disease was the same as the bacterial spot and canker of stone fruits caused by X. arboricola pv. pruni in the country. [ABSTRACT FROM AUTHOR]

Common bacterial blight caused by Xanthomonas axonopodis pv. phaseoli (Xap) is one of the most destructive diseases limiting the production of common bean (Phaseolus vulgaris) in Iran. The disease has previously been described on common bean and mung bean from several regions of Iran, including the central plain and south-western provinces. In this study, lima bean (Phaseolus lunatus cv. Christmas) plants are being reported as a new natural host of Xap in East Azerbaijan and West Azerbaijan Provinces, northwestern Iran. Disease symptoms consisted initially of watersoaked spots that progressed to irregular necrotic lesions with chlorotic margins. Infection was observed to affect up to 40% of plants in the field. Identification of the pathogen was based on the biochemical and molecular characteristics, as well as the pathogenicity tests. To our knowledge, this is the first report of X. axonopodis pv. phaseoli causing common bacterial blight on lima bean plants in Iran. [ABSTRACT FROM AUTHOR]

A new orange variant of Curtobacterium flaccumfaciens pv . flaccumfaciens was isolated from seeds of common bean cv. Daneshkadeh and Dehghan stored in the seed banks in Khomein Bean Research Station, and field plants (cv. Local Khomein) in Arak, Iran. The pathogenicity of the isolates was confirmed on 5- to 7-day-old seedlings of cv. Daneshkadeh. Marginal necrosis and interveinal chlorosis on first trifoliate leaves were observed 10-15 days after inoculation. Amplification of 306 bp fragment of orange-pigmented strains using Cff FOR2- and Cff REV4-specific prime pair characterized them as C. flaccumfaciens pv. flaccumfaciens. Although the yellow-pigmented variant of the causal agent was previously reported on cowpea, this is the first report of orange variant of C. flaccumfaciens pv. flaccumfaciens causing bacterial wilt on common bean in Iran. [ABSTRACT FROM AUTHOR]

During surveys in cowpea fields of Marand County, East Azerbaijan province, Iran, in the summer of 2013, a suspected bacterial disease was observed on cowpea leaves as tan spots and interveinal necrotic lesions surrounded by chlorotic margins. The disease was of high incidence where some fields had been fully destroyed and severity of the disease in some fields had reached up to 70%. Gram-positive, yellow-pigmented, coryneform bacteria were isolated from infected leaves. Pathogenicity of isolates was confirmed on 20-day-old cowpea (cv. Khoy) plants, and they were identified as Curtobacterium flaccumfaciens pv. flaccumfaciens based on biochemical test results confirmed using specific PCR primers. This is the first report of C. flaccumfaciens pv. flaccumfaciens, the causal agent of cowpea bacterial wilt in Iran. [ABSTRACT FROM AUTHOR]

Bacterial wilt, which is caused by Curtobacterium flaccumfaciens pv . flaccumfaciens ( Cff ), has recently emerged in common bean, lima bean, and cowpea plants in the provinces of East Azerbaijan and Markazi in the northwestern and central regions of Iran, respectively. Because the development of wilt-resistant common bean cultivars is the most cost-effective and lasting method for controlling the disease, this study was conducted to evaluate the common bean germplasms provided by the Khomein Bean Research Station (Khomein County, Markazi Province) for their resistance to Cff . Forty-three common bean genotypes were used in this study, and the node infiltration method was used to inoculate common bean seedlings at the V3 stage of development. Based on the disease severity index (DSI) values, the brown pinto ks21186 line, the red pinto Chili landrace, and the brown pinto ks21193 line were identified as resistant to the highly virulent yellow variant of the bacterial wilt pathogen Cffcb124 strain (DSI: 0.85, 0.9 and 0.92 out of 4.0, respectively). Among the three resistant germplasms, the ks21186 line was also resistant to other strains of the pathogen isolated from cowpea (Cffcw13 and Cffcw11), lima bean (Cffcb122), and common bean (Cffcb145 and Cffcb125) (mean DSI: 0.77). The mean DSI values of the ks21193 line and the Chili landrace were higher than the DSI threshold, below which indicates resistance (DSI: 1.14 and 1.22, respectively). This study concludes that a new bacterial wilt-resistant line (ks21186) exists among the common bean varieties at the Khomein Bean Research Station of Iran. [ABSTRACT FROM AUTHOR]

Summary. Rhizobium leguminosarum bv. phaseoli was evaluated for its capacity to trigger resistance to common bacterial blight (CBB) of common bean caused by Xanthomonas axonopodis pv. phaseoli (Xap), under greenhouse and field conditions. A common bean cultivar and three lines including two tolerant lines (Ks51103 and BF13607) and the susceptible cultivar Khomein and line Ks21479, were used. R. leguminosarum bv. phaseoli, was applied as a seed treatment and its effect on disease severity (DS) was compared with untreated (control) plants and with urea fertilizer treated plants in both the greenhouse and the field. R. leguminosarum bv. phaseoli in common bean roots tended to reduce CBB severity and to improve plant growth, particularly the 100-seed weight, in the field. The greatest decrease in CBB in the greenhouse occurred 15 days after inoculation (DAI) of Xap in the cultivar Khomein and line Ks21479, and 30 DAI in lines Ks51103 and BF13607. In the field, however, the greatest decrease occurred 35 DAI in the cultivar and all lines. Furthermore, in the field the greatest improvement in 100-seed weight occurred in the cultivar Khomein treated with R. leguminosarum bv. phaseoli. The potential of using R. leguminosarum bv. phaseoli in the management of CBB and the possible mechanisms that induce resistance in this symbiotic system are discussed. [ABSTRACT FROM AUTHOR]

Mycopathogenic bacteria play a pivotal role in the productivity of edible mushrooms grown under controlled conditions. In this study, we carried out a comprehensive farm survey and sampling (2018 to 2021) on button mushroom (Agaricus bisporus) farms in 15 provinces in Iran to monitor the status of bacterial pathogens infecting the crop. Mycopathogenic bacterial strains were isolated from pins, stems, and caps, as well as the casing layer on 38 mushroom farms. The bacterial strains incited symptoms on mushroom caps ranging from faint discoloration to dark brown and blotch of the inoculated surfaces. Among the bacterial strains inciting disease symptoms on bottom mushroom, 40 were identified as Ewingella americana based on biochemical assays and phylogeny of 16S rRNA and the gyrB gene. E. americana strains differed in their aggressiveness on mushroom caps and stipes, where the corresponding symptoms ranged from deep yellow to dark brown. In the phylogenetic analyses, all E. americana strains isolated in this study were clustered in a monophyletic clade closely related to the nonpathogenic and environmental strains of the species. BOX-PCR-based fingerprinting revealed intraspecific diversity. Using the cutoff level of 73 to 76% similarity, the strains formed six clusters. A chronological pattern was observed, where the strains isolated in 2018 were differentiated from those isolated in 2020 and 2021. Taken together, due to the multifaceted nature of the pathogen, such a widespread occurrence of E. americana on mushroom farms in Iran could be an emerging threat for the mushroom industry in the country. [ABSTRACT FROM AUTHOR]

Agrobacterium tumefaciens species complex contains a set of diverse bacterial strains, most of which are well known for their pathogenicity on agricultural plants causing crown gall diseases. Members of A. tumefaciens species complex are classified into several taxonomically distinct lineages called “genomospecies” (13 genomospecies until early 2021). Recently, two genomospecies, G19 (strains RnrT, Rew, and Rnw) and G20 (strains OT33T and R13) infecting Rosa sp. plants in Iran, were described based on biochemical and molecular-phylogenetic data. Whole genome sequence-based core-genome phylogeny followed by average nucleotide identity (ANI) and digital DNA-DNA hybridization (dDDH) calculations performed in this study suggested that genomospecies G19 and G20 could be described as two novel and standalone species. In the phylogenetic tree, these two new genomospecies were clustered separately from other genomospecies/species of A. tumefaciens species complex. Moreover, both ANI and dDDH indices between the G19/G20 strains and other Rhizobiaceae members are clearly below the accepted thresholds for prokaryotic species description. Hence, Agrobacterium burrii sp. nov. is proposed to encompass the G19 strains, with RnrT = CFBP 8705T = DSM 112541T as type strain. Agrobacterium shirazense sp. nov. is also proposed to include G20 strains, with OT33T = CFBP 8901T = DSM 112540T as type strain. [ABSTRACT FROM AUTHOR]

Bacterial wilt and brown rot disease caused by Ralstonia solanacearum species complex (RSSC) is one of the major constraints of potato (,Solarium tuberosum) production around the globe. During 2017 to 2018, an extensive field survey was conducted in six potato-growing provinces of Iran to monitor the status of bacterial wilt disease. Pathogenicity and host range assays using 59 bacterial strains isolated in Iran showed that they were pathogenic on eggplant, red nightshade, pepper, potato and tomato, while nonpathogenic on common bean, cowpea, cucumber, sunflower, zinnia and zucchini. PCR-based diagnosis revealed that the strains belong to the phylotype IIB/sequevar 1 (IIB/I) lineage of the RSSC. Furthermore, a fivegene multilocus sequence analysis and typing (egl, fliC, gyrB, mutS, and rplB) confirmed the phylogenetically near-homogeneous nature of the strains within IIB/I lineage. Four sequence types were identified among 58 IIB/1 strains isolated in Iran. Phylogenetically near-homogeneous nature of the strains in Iran raise questions about the mode of inoculum entry of the bacterial wilt pathogen into the country (one-time introduction versus multiple introductions), while the geographic origin of the Iranian R. solanacearum strains remains undetermined. Furthermore, sequence typing showed that there were shared alleles (haplotypes) and sequence types among the strains isolated in geographically distant areas in Iran, suggesting intranational transmission of the pathogen in the country. [ABSTRACT FROM AUTHOR]

In this study, we explored the pathogenicity and phylogenetic position of Agrobacterium spp. strains isolated from crown gall tissues on annual, perennial, and ornamental plants in Iran. Of the 43 strains studied, 10 strains were identified as Allorhizobium vitis (formerly Agrobacterium vitis) using the species-specific primer pair PGF/PGR. Thirty-three remaining strains were studied using multilocus sequence analysis of four housekeeping genes (i.e., atpD, gyrB, recA, and rpoB), from which seven strains were identified as A. larrymoorei and one strain was identified as A. rubi (Rer); the remaining 25 strains were scattered within the A. tumefaciens species complex. Two strains were identified as genomospecies 1 (Gl), seven strains were identified as A. radiobcicter (G4), seven strains were identified as A. deltaense (G7), two strains were identified as A. nepotum (G14), and one strain was identified as “A. viscosum” (G15). The strains Rnr, Rnw, and Rew as well as the two strains OT33 and R13 all isolated from rose and the strain Apl isolated from apple were clustered in three atypical clades within the A. tumefaciens species complex. All but eight strains (i.e., NeclO, Ph38, Ph49, fic9, Fic72, R13, OT33, and Apl) were pathogenic on tomato and sunflower seedlings in greenhouse conditions, whereas all but three strains (i.e., fic9, Fic72, and OT33) showed tumorigenicity on carrot root discs. The phylogenetic analysis and nucleotide diversity statistics suggested the existence of two novel genomospecies within the A. tumefaciens species complex, which we named “G19” and “G20.” Hence, we propose the strains Rew, Rnw, and Rnr as the members of “G19” and the strains R13 and OT33 as the members of G20, whereas the phylogenetic status of the atypical strain Apl remains undetermined. [ABSTRACT FROM AUTHOR]

This study provides a phylogeographic insight into the population diversity of Xanthomonas translucens strains causing bacterial leaf streak disease of small-grain cereals in Iran. Among the 65 bacterial strains isolated from wheat, barley, and gramineous weeds in eight Iranian provinces, multilocus sequence analysis and typing (MLSA and MLST) of four housekeeping genes (dnaK, fyuA, gyrB, and rpoD), identified 57 strains as X. translucens pv. undulosa, while eight strains were identified as X. translucens pv. translucens. Although the pathogenicity patterns on oat and ryegrass weed species varied among the strains, all X. translucens pv. undulosa strains were pathogenic on barley, Harding's grass, rye (except for XtKm35) and wheat, and all X. translucens pv. translucens strains were pathogenic on barley and Harding's grass, while none of the latter group was pathogenic on rye or wheat (except for XtKm18). MLST using the 65 strains isolated in Iran, as well as the sequences of the four genes from 112 strains of worldwide origin retrieved from the GenBank database, revealed higher genetic diversity (i.e., haplotype frequency, haplotype diversity, and percentage of polymorphic sites) among the Iranian population of X. translucens than among the North American strains of the pathogen. High genetic diversity of the BLS pathogen in Iran was in congruence with the fact that the Iranian Plateau is considered the center of origin of cultivated wheat. However, further studies using larger collections of strains are warranted to precisely elucidate the global population diversity and center of origin of the pathogen. [ABSTRACT FROM AUTHOR]

Bacterial blight and leaf spot of geraniums is a destructive disease of cultivated Pelargonium species around the world. During 2020 to 2021, surveys were conducted in seven geranium-growing provinces of Iran to monitor the status of bacterial blight and leaf spot disease. The disease was observed in six surveyed provinces varying in the extent of occurrence and severity. Twenty-two gram-negative pale-yellow bacterial strains resembling members of Xanthomonas were isolated from symptomatic leaves and stems. Pathogenicity and host range assays showed that the bacterial strains were pathogenic on Pelargonium grandiflorum , P. graveolens , P. peltatum , and P. zonale . All strains were positive for a PCR test using the primer pair XcpM1/XcpM2, which is specific for X. hortorum pv. pelargonii . Phylogenetic analysis using the sequences of gyrB and lepA genes showed that the 22 strains clustered in a clade among the sequences of X. hortorum pv. pelargonii strains retrieved from GenBank but were distinct from the other pathovars of X. hortorum . BOX-PCR-based fingerprinting using BOX-A1R primer revealed that the strains isolated in this study were grouped into two clusters, while no distinct correlation was observed between the host/area of isolation and BOX-PCR fingerprinting. None of the strains obtained in this study nor the reference strain of the pathogen produced bacteriocin against each other. Results obtained in this study shed light on the geographic distribution, taxonomic status, and host range of the bacterial blight and leaf spot pathogen of geraniums in Iran, paving the path for further research on disease management., Competing Interests: The author(s) declare no conflict of interest.

Taxonomically diverse Pseudomonas species induce bacterial blotch of edible mushrooms around the world. Pseudomonas tolaasii , [ P . gingeri ], and P. agarici are dominant mycopathogenic pseudomonads in mushroom production farms. In this study, among 216 mycopathogenic bacterial strains isolated from edible mushrooms in Iran, 96 strains were identified as Pseudomonas spp., while only three strains were preliminarily identified as P. agarici . Multilocus sequence analysis showed that only one strain (FH2) authentically belonged to P. agarici , while the other two strains either belonged to [ P. gingeri ] or represented a unique phylogenetic clade. The three strains also differed from each other in phenotypic characteristics, for example, production of fluorescent pigment and the reaction to tolaasin produced by P. tolaasii . Pathogenicity assays under a controlled environment showed that the symptoms induced by authentic P. agarici were far less severe than those caused by the predominant species P. tolaasii . Furthermore, coinoculation of P. agarici with three bacterial pathogens that are prevalent in Iran on mushroom caps, that is, P. tolaasii , Ewingella americana , and Mycetocola sp., resulted in the development of combined symptoms representing characteristics of both pathogens. The antibiosis assay showed that tolaasin-producing strains of P. tolaasii could inhibit the growth of P. agarici , while tolaasin-negative strains of the same species were unable to do so. This led us to the hypothesis that the inhibitory effect of P. tolaasii on P. agarici is driven by tolaasin production in the former species. This inhibitory effect is also associated with the rarity of P. agarici in natural conditions., Competing Interests: The author(s) declare no conflict of interest.

Ralstonia solanacearum , the causal agent of bacterial wilt and brown rot disease, is one of the major pathogens of solanaceous crops, including potato, around the globe. Biovar 2T (phylotype II/sequevar 25) of R. solanacearum is adapted to tropical lowlands and is only reported in South America and Iran. Thus far, no genome resource of the biovar 2T of the pathogen has been available. Here, we present the near-complete genome sequences of the biovar 2T strain CFBP 8697 as well as strain CFBP 8695 belonging to biovar 2 race 3, both isolated from potato in Iran. The genomic data of biovar 2T will extend our understanding of the virulence features of R. solanacearum and pave the way for research on biovar 2T functional and interaction genetics.

From September to December 2018, commercial button mushroom ( Agaricus bisporus ) farms in central Iran were surveyed to monitor the causal agent(s) of browning and blotch symptoms on mushroom caps. In addition to dozens of pseudomonads (i.e., Pseudomonas tolaasii and Pseudomonas reactans ), six slow-growing gram-positive bacterial strains were isolated from blotched mushroom caps. These bacteria presented as creamy white, circular, smooth, nonfluorescent, and shiny colonies with whole margins resembling members of Microbacteriaceae (Actinobacteria). All of the actinobacterial strains were aggressively pathogenic on cut cap surface of two edible mushrooms (i.e., A. bisporus and Pleurotus eryngii ), inducing brown pit symptoms 48 h postinoculation. The strains did not induce symptoms on the vegetables tested (i.e., carrot, cucumber, and potato), and they did not affect the growth of mycelium of tested plant-pathogenic fungi (i.e., Acremonium sp., Fusarium spp., and Phytopythium sp.). Phylogeny of 16S ribosomal RNA and multilocus sequence analysis of six housekeeping genes (i.e., atpD, dnaK, gyrB, ppK, recA, and rpoB) revealed that the bacterial strains belong to the actinobacterial genus Mycetocola spp., whereas the species status of the strains remains undetermined. Mushroom-associated Mycetocola species were previously reported to be capable of detoxifying tolaasin, a toxin produced by P. tolaasii , whereas the strains isolated in this study did not show tolaasin detoxification activities. Altogether, this is the first report of a mushroom disease caused by an actinobacterial species, and "bacterial brown pit" was assigned as the common name of the disease.

Bacterial leaf streak caused by different pathovars of Xanthomonas translucens is the most important seedborne bacterial disease of small grain cereals. However, variations in the virulence-associated genomic areas of the pathogen remain uninvestigated. In this study, the diversity of transcription activator-like effectors (TALE) was investigated using the Southern blotting of Bam HI - digested genomic DNAs in the Iranian strains of X. translucens . All 65 X. translucens strains were assigned into 13 genotypes, where 57 X. translucens pv. undulosa strains were placed in genotypes 1 to 8, and seven X. translucens pv. translucens strains were placed in genotypes 9 to 12. Interestingly, we did not find any TALE genes in the strain XtKm7 (genotype 13), which showed to be pathogenic only on barley. Virulence and aggressiveness of these strains in greenhouse conditions were in agreement with the TALE-based clustering of the strains in the pathovar level, though variations were observed in the aggressiveness of X. translucens pv. undulosa strains. In general, strains containing higher numbers of putative TALE genes were more virulent on wheat and barley than strains containing fewer. This is the first TALE-based genetic diversity analysis on X. translucens strains and provides novel insights into the virulence repertories and genomic characteristics of the pathogen. Further investigations using TALE mutagenesis and complementation analysis are warranted to precisely elucidate the role of each detected X. translucens TALE in bacterial virulence and aggressiveness either on wheat or barley.

In this study, we provide a polyphasic characterization of 18 Pseudomonas spp. strains associated with alfalfa leaf spot symptoms in Iran. All of the strains were pathogenic on alfalfa, although the aggressiveness and symptomology varied among the strains. All strains but one were pathogenic on broad bean, cucumber, honeydew, and zucchini, whereas only a fraction of the strains were pathogenic on sugar beet, tomato, and wheat. Syringomycin biosynthesis genes ( syrB1 and syrP ) were detected using the corresponding PCR primers in all of the strains isolated from alfalfa. Phylogenetic analyses using the sequences of four housekeeping genes ( gapA , gltA , gyrB , and rpoD ) revealed that all of the strains except one (Als34) belong to phylogroup 2b of P. syringae sensu lato , whereas strain Als34 placed within phylogroup 1 close to the type strain of P. syringae pv. apii . Among the phylogroup 2b strains, nine strains were phylogenetically close to the P. syringae pv. aptata clade, whereas the remainder were scattered among P. syringae pv. atrofaciens and P. syringae pv. syringae strains. Pathogenicity and host range assays of the bacterial strains evaluated in this study on a set of taxonomically diverse plant species did not allow us to assign a "pathovar" status to the alfalfa strains. However, these results provide novel insight into the host range and phylogenetic position of the alfalfa-pathogenic members of P. syringae sensu lato , and they reveal that phenotypically and genotypically heterogeneous strains of the pathogen cause bacterial leaf spot of alfalfa.

Tomato bacterial canker caused by Clavibacter michiganensis subsp. michiganensis is one of the most important seed-borne tomato diseases around the globe. The disease was initially reported in 1993 in Iran, and it became a rising threat for the multibillion dollar tomato industry of the country during the last decade. In this study, using phylogeographic analyses, we determined genetic diversity and geographic distribution of C. michiganensis subsp . michiganensis in Iran. Our field surveys showed that the pathogen is expanding into the southern and eastern areas of the country. Furthermore, multilocus sequence analysis and typing (MLSA/MLST) using the sequences of five housekeeping genes ( atpD , gyrB , ppk , recA , and rpoB ) revealed that 37 C. michiganensis subsp . michiganensis strains isolated in Iran had high genetic diversity and placed in 15 sequence types (STs), while all the available 184 worldwide C. michiganensis subsp . michiganensis sequences were placed in 43 STs. MLSA divided the worldwide C. michiganensis subsp . michiganensis strains into two phylogroups (I and II). Among the 37 strains isolated in Iran, 30 strains clustered in phylogroup I, while 7 strains clustered in phylogroup II. Phylogeographic data inferred from the allelic profile of the five housekeeping genes suggested multiple introductions of C. michiganensis subsp . michiganensis inoculum into Iran, while the geographic origin of the Iranian C. michiganensis subsp . michiganensis strains remains undetermined. Further analyses using higher numbers of strains are warranted to decipher the evolutionary history of C. michiganensis subsp . michiganensis in Iran. Additionally, stricter seed/transplant inspections are recommended to reduce the risk of pathogen expansion to areas with no history of the disease. IMPORTANCE Clavibacter michiganensis subsp. michiganensis , the causal agent of tomato bacterial canker disease, is one of the economically important pathogens of solanaceous crops (e.g., eggplant, pepper, and tomato) around the world. The disease occurs in many countries, with a particular importance in regions characterized by high precipitation and humid environmental conditions. As a seed-borne pathogen, C. michiganensis is included in the A2 (high risk) list of quarantine pathogens by the European and Mediterranean Plant Protection Organization (EPPO). Bacterial canker disease was reported for the first time in 1993 in Iran, while the geographic distribution, genetic diversity, and phylogenetic position of the causal agent remain undetermined. In this study, using the multilocus sequence analysis and typing (MLSA/MLST) approach, we provided a phylogeographic scheme for the . michiganensis is included in the A2 (high risk) list of quarantine pathogens by the European and Mediterranean Plant Protection Organization (EPPO). Bacterial canker disease was reported for the first time in 1993 in Iran, while the geographic distribution, genetic diversity, and phylogenetic position of the causal agent remain undetermined. In this study, using the multilocus sequence analysis and typing (MLSA/MLST) approach, we provided a phylogeographic scheme for the C. michiganensis strains isolated in Iran. Furthermore, global-scale phylogenetic analyses led to determination of phylogenetic position of Iranian . michiganensis strains isolated in Iran. Furthermore, global-scale phylogenetic analyses led to determination of phylogenetic position of Iranian C. michiganensis strains among worldwide population of the pathogen. Based on diversity parameters and population structure, we suggest relatively higher genetic diversity of the bacterial canker pathogen in Iran than has so far been observed in the other areas of the world. Results obtained in this study provide a novel insight into the genetic diversity and population structure of the bacterial canker pathogen on a global scale.. michiganensis strains among worldwide population of the pathogen. Based on diversity parameters and population structure, we suggest relatively higher genetic diversity of the bacterial canker pathogen in Iran than has so far been observed in the other areas of the world. Results obtained in this study provide a novel insight into the genetic diversity and population structure of the bacterial canker pathogen on a global scale., (Copyright © 2019 American Society for Microbiology.)