scholarly journals Pseudomonas entomophila 23S Produces a Novel Antagonistic Compound against Clavibacter michiganensis subsp. michiganensis, a Pathogen of Tomato Bacterial Canker

2021 ◽  
Vol 1 (1) ◽  
pp. 60-73
Author(s):  
Yoko Takishita ◽  
Alfred Souleimanov ◽  
Carine Bourguet ◽  
Leanne B. Ohlund ◽  
Alexandre A. Arnold ◽  
...  

Biopesticides, which consist of antagonistic microorganisms, or the bioactive compounds they produce offer attractive alternatives to synthetic agrochemicals as a means of reducing plant diseases in crop production systems. In our previous study, a strain of Pseudomonas entomophila 23S was reported to have a strong antagonistic activity toward Clavibacter michiganensis subsp. michiganensis (Cmm), a pathogen of tomato bacterial canker disease, which causes serious damage to the tomato production industry. This study focused on purification and characterization of the anti-Cmm compound(s) produced by P. entomophila 23S. The cell-free supernatant was first subjected to butanol extraction, fractioned by solid-phase extraction (SPE), and purified by high-performance liquid-chromatography (HPLC). The semi-preparative purified compounds retained anti-Cmm activity. These compounds were then characterized by liquid chromatography coupled with high-resolution tandem mass spectrometry (LC-HRMS/MS) and nuclear magnetic resonance (NMR). Two anti-Cmm compounds, C15H16N2O and C16H18N2O, were isolated by this procedure. From the LC/MS spectra, the compounds were predicted to have an isoquinoline ring, the presence of which was confirmed by NMR. The compounds differed with respect to the presence of a methyl group in a side chain. In addition, P. entomophila 23S grown in different culture media produced the greatest amounts of these anti-Cmm compounds when in Nutrient Broth (NB), where bacterial growth was the slowest. NB composition is more favorable in stimulating P. entomophila 23S to produce higher amounts of these anti-Cmm compounds.

Plant Disease ◽  
2004 ◽  
Vol 88 (6) ◽  
pp. 680-680 ◽  
Author(s):  
A. Anwar ◽  
P. S. van der Zouwen ◽  
S. Ilyas ◽  
J. M. van der Wolf

In 2002, Clavibacter michiganensis subsp. michiganensis (Smith) Davis, the causal organism of bacterial canker of tomato (Lycopersicon esculentum), was isolated from two of six commercial asymptomatic tomato seed lots produced on Java in Indonesia. C. michiganensis subsp. michiganensis has not been reported in Indonesia previously. Methods based on the protocol of the International Seed Health Initiative were used to extract and identify the presence of C. michiganensis subsp. michiganensis in tomato seed. C. michiganensis subsp. michiganensis was isolated with dilution plating on the semiselective media D2ANX and mSCM. The identity of the colonies was confirmed by immunofluorescence microscopy, polymerase chain reaction (2), fatty methyl ester analysis, enzyme-linked immunosorbent assay based on monoclonal antibody 103 (1), and a pathogenicity test in which three replicate tomato plants were stem inoculated with 108 cells ml-1. Within 2 weeks, stripes on stems developed that split and exposed reddish brown cavities (stem cankers). The presence of C. michiganensis subsp. michiganensis poses a direct threat on tomato production, which is one of five economically most important vegetable crops in Indonesia. References: (1) A. Alvarez et al. Phytopathology 83:1405, 1993. (2) M. S. Santos et al. Seed Sci. Technol. 25:581, 1997.


2015 ◽  
Vol 65 (Pt_3) ◽  
pp. 811-826 ◽  
Author(s):  
Jarred Yasuhara-Bell ◽  
Anne M. Alvarez

The genus Clavibacter contains one recognized species, Clavibacter michiganensis . Clavibacter michiganensis is subdivided into subspecies based on host specificity and bacteriological characteristics, with Clavibacter michiganensis subsp. michiganensis causing bacterial canker of tomato. Clavibacter michiganensis subsp. michiganensis is often spread through contaminated seed leading to outbreaks of bacterial canker in tomato production areas worldwide. The frequent occurrence of non-pathogenic Clavibacter michiganensis subsp. michiganensis -like bacteria (CMB) is a concern for seed producers because Clavibacter michiganensis subsp. michiganensis is a quarantine organism and detection of a non-pathogenic variant may result in destruction of an otherwise healthy seed lot. A thorough biological and genetic characterization of these seed-associated CMB strains was performed using standard biochemical tests, cell wall analyses, metabolic profiling using Biolog, and single-gene and multilocus sequence analyses. Combined, these tests revealed two distinct populations of seed-associated members of the genus Clavibacter that differed from each other, as well as from all other described subspecies of Clavibacter michiganensis . DNA–DNA hybridization values are 70 % or higher, justifying placement into the single recognized species, C. michiganensis , but other analyses justify separate subspecies designations. Additionally, strains belonging to the genus Clavibacter isolated from pepper also represent a distinct population and warrant separate subspecies designation. On the basis of these data we propose subspecies designations for separate non-pathogenic subpopulations of Clavibacter michiganensis : Clavibacter michiganensis subsp. californiensis subsp. nov. and Clavibacter michiganensis subsp. chilensis subsp. nov. for seed-associated strains represented by C55T ( = ATCC BAA-2691T = CFBP 8216T) and ZUM3936T ( = ATCC BAA-2690T = CFBP 8217T), respectively. Recognition of separate subspecies is essential for improved international seed testing operations.


2008 ◽  
Vol 69 (1) ◽  
pp. 125-134 ◽  
Author(s):  
Czesław Ślusarski

Attempts at Biological Control ofClavibacter michiganensissubsp.michiganensisOn Rockwool-Grown Greenhouse TomatoesTwo greenhouse experiments were conducted in which tomato plants artificially inoculated withClavibacter michiganensissubsp.michiganensis(Cmm) were grown in an open rockwool system as spring and autumn crops. Two isolates of the rhizosphere bacteria,Pseudomonas fluorescensstrain PSR21,Pseudomonas reactansstrain GGS14, a commercial biocontrol agent Aqua Bac Plus (Bacillusspp.) and a proprietary disinfectant containing QAC+Chx, applied at weekly intervals, were evaluated for their efficiency in the suppression of the bacterial canker of tomato. All treatments tested revealed to be ineffective in controlling the disease. The introduction ofCmmbacteria into the fresh rockwool in the first year of its usage resulted in a 100% death of tomato plants, whereas following an artificial inoculation of two- and three-year-old rockwool slabs withCmmbacteria dead plants amounted to 70 and 58%, respectively. This indicates that in the re-used rockwool a natural microbial suppressiveness to bacterial canker of tomato might be developed in the root zone.


Plant Disease ◽  
2008 ◽  
Vol 92 (4) ◽  
pp. 649-649 ◽  
Author(s):  
R. Ftayeh ◽  
A. von Tiedemann ◽  
B. Koopmann ◽  
K. Rudolph ◽  
M. Abu-Ghorrah

Between March and mid April of 2007, several extensive surveys for Clavibacter michiganensis subsp. michiganensis were carried out among greenhouses in the coastal strip provinces of the Mediterranean Sea in north-west Syria (Latakia and Tartous), where a large proportion of Syrian fresh-market tomatoes are produced. This bacterium causes bacterial canker of tomato and is considered an A2 quarantine pathogen by the European Plant Protection Organization (EPPO). It is currently present in all major tomato-production areas in the EPPO region (4), but has not been previously reported in Syria. The survey revealed typical canker symptoms in 7% of 150 inspected greenhouses that contained cvs. Dima, Huda, and Astona. These symptoms included stunting, dark brown-to-black lesions on the leaf margins, wilting and defoliation of whole plants, and vascular discoloration. The disease incidence in such greenhouses was estimated at 15% at the time of the survey. Diseased plants were surface sterilized and homogenized in sterile water. Serial dilutions were plated on nutrient glucose agar. Suspected colonies were further purified by repeated restreaking on new agar plates. All 10 of the suspected strains obtained from different locations were identified as C. michiganensis subsp. michiganensis on the basis of the following observations: bacterial cells of all strains had a coryneform shape, were nonmotile, gram positive according to Gram's reaction test with 3% KOH (2), oxidase-negative, and caused hypersensitive reactions on leaves of Mirabilis jalaba (1) within 24 h. PCR assays were conducted with the C. michiganensis subsp. michiganensis-specific primer set PSA-4/R (3) and template DNA prepared from in-vitro-grown bacteria with the MasterPure Gram Positive DNA Purification Kit (Epicentre Biotechnologies, Madison, WI). The expected 270-bp amplicon was observed for both reference strains as well as the Syrian strains. Pathogenicity of the strains was confirmed by artificial inoculation of 6-week-old tomato plants (Lycopersicon esculentum Mill. cv. Lyconorma). Inoculation was performed by stabbing the stem with a sterile needle through a drop (~35 μl) of bacterial suspension (~108 CFU/ml in 0.01 M MgSO4) placed in the axil of the second or third true leaf. Three tomato seedlings were inoculated with each strain. Control plants were inoculated with sterile 0.01 M MgSO4. Symptoms including lateral wilt of leaflets, stem lesions, and wilting of whole plants were observed within 10 to 15 days after inoculation, except for the negative control. To fulfill Koch's postulates, reisolation and reidentification of the pathogen was conducted as previously described. To our knowledge, this is the first record of the occurrence of bacterial canker of tomato in Syria. References: (1) R. D. Gitaitis. Plant Dis. 74:58, 1990. (2) T. J. Gregersen. Appl. Microbiol. Biotechnol. 5:123, 1978. (3) K. H. Pastrik and F. A. Rainey. J. Phytopathol. 147:687, 1999. (4) I. M. Smith and L. M. F. Charles, eds. Map 253 in: Distribution Maps of Quarantine Pests for Europe. EPPO/CABI, 1998.


2020 ◽  
Vol 8 (11) ◽  
pp. 1679
Author(s):  
Valentina Méndez ◽  
Miryam Valenzuela ◽  
Francisco Salvà-Serra ◽  
Daniel Jaén-Luchoro ◽  
Ximena Besoain ◽  
...  

The genus Clavibacter has been associated largely with plant diseases. The aims of this study were to characterize the genomes and the virulence factors of Chilean C. michiganensis subsp. michiganensis strains VL527, MSF322 and OP3, and to define their phylogenomic positions within the species, Clavibacter michiganensis. VL527 and MSF322 genomes possess 3,396,632 and 3,399,199 bp, respectively, with a pCM2-like plasmid in strain VL527, with pCM1- and pCM2-like plasmids in strain MSF322. OP3 genome is composed of a chromosome and three plasmids (including pCM1- and pCM2-like plasmids) of 3,466,104 bp. Genomic analyses confirmed the phylogenetic relationships of the Chilean strains among C.michiganensis subsp. michiganensis and showed their low genomic diversity. Different virulence levels in tomato plants were observable. Phylogenetic analyses of the virulence factors revealed that the pelA1 gene (chp/tomA region)—that grouped Chilean strains in three distinct clusters—and proteases and hydrolases encoding genes, exclusive for each of the Chilean strains, may be involved in these observed virulence levels. Based on genomic similarity (ANIm) analyses, a proposal to combine and reclassify C. michiganensis subsp. phaseoli and subsp. chilensis at the species level, as C. phaseoli sp. nov., as well as to reclassify C. michiganensis subsp. californiensis as the species C. californiensis sp. nov. may be justified.


2016 ◽  
Vol 66 (10) ◽  
pp. 4065-4070 ◽  
Author(s):  
Eom-Ji Oh ◽  
Chungyun Bae ◽  
Han-Beoyl Lee ◽  
In Sun Hwang ◽  
Hyok-In Lee ◽  
...  

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