scholarly journals Comparative Genomics of Pathogenic Clavibacter michiganensis subsp. michiganensis Strains from Chile Reveals Potential Virulence Features for Tomato Plants

2020 ◽  
Vol 8 (11) ◽  
pp. 1679
Author(s):  
Valentina Méndez ◽  
Miryam Valenzuela ◽  
Francisco Salvà-Serra ◽  
Daniel Jaén-Luchoro ◽  
Ximena Besoain ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Phylogenetic Analyses ◽  
Plant Diseases ◽  
Genomic Diversity ◽  
Clavibacter Michiganensis ◽  
Tomato Plants ◽  
Clavibacter Michiganensis Subsp ◽  
Genomic Analyses ◽  
Encoding Genes ◽  
Genomic Similarity

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.

Attempts at Biological Control of Clavibacter michiganensis subsp. michiganensis On Rockwool-Grown Greenhouse Tomatoes

2008 ◽  
Vol 69 (1) ◽  
pp. 125-134 ◽  
Author(s):  
Czesław Ślusarski
Keyword(s):  
Biological Control ◽  
Biocontrol Agent ◽  
Root Zone ◽  
First Year ◽  
Bacterial Canker ◽  
Clavibacter Michiganensis ◽  
Tomato Plants ◽  
Clavibacter Michiganensis Subsp ◽  
Greenhouse Tomatoes ◽  
Bacterial Canker Of Tomato

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.

scholarly journals First Record of Clavibacter michiganensis subsp. michiganensis Causing Canker of Tomato Plants in Syria

Plant Disease ◽  
2008 ◽  
Vol 92 (4) ◽  
pp. 649-649 ◽  
Author(s):  
R. Ftayeh ◽  
A. von Tiedemann ◽  
B. Koopmann ◽  
K. Rudolph ◽  
M. Abu-Ghorrah
Keyword(s):  
First Record ◽  
Bacterial Suspension ◽  
Bacterial Canker ◽  
Clavibacter Michiganensis ◽  
Fresh Market ◽  
Gram Positive ◽  
Tomato Plants ◽  
Clavibacter Michiganensis Subsp ◽  
Whole Plants ◽  
Bacterial Canker Of Tomato

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.

EFFECT OF ESSENTIAL OILS IN THE CONTROL OF THE Clavibacter michiganensis SUBESPECIE michiganensis IN TOMATO (Lycopersicum esculentum L.) PLANTS

Biotecnia ◽  
2018 ◽  
Vol 20 (3) ◽  
pp. 96-101 ◽  
Author(s):  
Jesús Borboa Flores ◽  
Jesús Ortega García ◽  
Francyelli Regina Costa Becheleni ◽  
Arnoldo Villegas Espinoza ◽  
Francisco Javier Wong-Corral ◽  
...  
Keyword(s):  
Essential Oil ◽  
Essential Oils ◽  
Antimicrobial Properties ◽  
Thymus Vulgaris ◽  
Clavibacter Michiganensis ◽  
Origanum Vulgare ◽  
Cinnamomum Zeylanicum ◽  
Tomato Plants ◽  
Clavibacter Michiganensis Subsp ◽  
Bactericidal Properties

Plants produce compounds with antimicrobial properties that can be used to control diseases of horticultural products, for this reason, the aim of this study was to enlarge the knowledge on the effect of essential oils as an alternative for control for bacterial canker Clavibacter michiganensis subsp. michiganensis (Smith) (Cmm). Essential oil of Lippia palmeri was obtained by hydridestillation from plants collected in Sonora, Mexico. Essential oils of Origanum vulgare, Thymus vulgaris and Cinnamomum zeylanicum were obtained from commercial stores in Hermosillo, Sonora, Mexico. To evaluate the effect of the essential oils on the bacterium Cmm, direct applications in two concentrations (1:1 and 1:5 (v/v)) on tomato plants foliage were carried out. Then, plants were inoculated with 1x10-7 CFU/mL of the bacterium and observed for 12 days. The evaluated parameters were incidence and severity. The essential oils and chemical compounds of Origanum vulgare and Thymus vulgaris had effect against Clavibacter michiganensis subsp. michiganensis up to 9 days after application in a 1:1 (v/v) concentration and up to 12 days after application with the 1:5 (v/v) concentration. The essential oil of Lippia palmeri only had effect until six days after the application with both concentrations; that is to say, only these three types of essential oils had bactericidal properties, since with the essential oil of Cinnamomum zeylanicum the inhibition of Cmm was very limited, both in incidence as in severity.

scholarly journals Preparation and use of monoclonal antibodies  to detect Phytophthora nicotianae  var. nicotianae

1999 ◽  
Vol 35 (No. 2) ◽  
pp. 41-46
Author(s):  
B. Pekárová-Kyněrová ◽  
M. Kutíková
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Fusarium Oxysporum ◽  
Indirect Elisa ◽  
Phytophthora Nicotianae ◽  
Clavibacter Michiganensis ◽  
Tomato Plants ◽  
Clavibacter Michiganensis Subsp

A monoclonal antibody (MAb 18) was prepared against purified mycelial proteins from Phytophthora nicotianae var. nicotianae. The specificity of MAb 18 (lgG class) was tested using indirect ELISA (PTA-ELISA).It cross-reacted with Phytophthora cacto­ rum, P. cinrzamomi, P. cryptogea, P. fragariae) but not with other fungi (Fusarium  oxysporum, Pythium ultimwn and P. oligan­ drwn) and bacteria (Clavibacter michiganensis subsp. michiganensis) isolated from tomato. Phytophthora nicotianae var. nicotianae was detected in roots and basal stems of artificially infected young tomato plants using indirect ELISA and immunoprinting.

Studies on the colonization of axenically grown tomato plants by a GFP-tagged strain of Clavibacter michiganensis subsp. michiganensis

2013 ◽  
Vol 139 (1) ◽  
pp. 53-66 ◽  
Author(s):  
Flávia M. Vieira Lelis ◽  
Robert Czajkowski ◽  
Ricardo Magela de Souza ◽  
Daniel H. Ribeiro ◽  
Jan M. van der Wolf
Keyword(s):  
Clavibacter Michiganensis ◽  
Tomato Plants ◽  
Clavibacter Michiganensis Subsp

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 ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Crop Production ◽  
Solid Phase ◽  
Culture Media ◽  
Plant Diseases ◽  
Bacterial Canker ◽  
Clavibacter Michiganensis ◽  
Tomato Production ◽  
Clavibacter Michiganensis Subsp ◽  
Antagonistic Microorganisms

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.

scholarly journals Colonization and Movement of GFP-Labeled Clavibacter michiganensis subsp. michiganensis During Tomato Infection

2012 ◽  
Vol 102 (1) ◽  
pp. 23-31 ◽  
Author(s):  
L. Chalupowicz ◽  
E.-M. Zellermann ◽  
M. Fluegel ◽  
O. Dror ◽  
R. Eichenlaub ◽  
...  
Keyword(s):  
Virulence Factors ◽  
Laser Scanning ◽  
Fluorescent Protein ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Wall Thickening ◽  
Apical Region ◽  
In Planta ◽  
Clavibacter Michiganensis ◽  
Clavibacter Michiganensis Subsp

The vascular pathogen Clavibacter michiganensis subsp. michiganensis is responsible for bacterial wilt and canker of tomato. Pathogenicity of this bacterium is dependent on plasmid-borne virulence factors and serine proteases located on the chromosomal chp/tomA pathogenicity island (PAI). In this study, colonization patterns and movement of C. michiganensis subsp. michiganensis during tomato infection was examined using a green fluorescent protein (GFP)-labeled strain. A plasmid expressing GFP in C. michiganensis subsp. michiganensis was constructed and found to be stable in planta for at least 1 month. Confocal laser-scanning microscopy (CLSM) of inoculated stems showed that the pathogen extensively colonizes the lumen of xylem vessels and preferentially attaches to spiral secondary wall thickening of the protoxylem. Acropetal movement of the wild-type strain C. michiganensis subsp. michiganensis NCPPB382 (Cmm382) in tomato resulted in an extensive systemic colonization of the whole plant reaching the apical region after 15 days, whereas Cmm100 (lacking the plasmids pCM1 and pCM2) or Cmm27 (lacking the chp/tomA PAI) remained confined to the area surrounding of the inoculation site. Cmm382 formed biofilm-like structures composed of large bacterial aggregates on the interior of xylem walls as observed by CLSM and scanning electron microscopy. These findings suggest that virulence factors located on the chp/tomA PAI or the plasmids are required for effective movement of the pathogen in tomato and for the formation of cellular aggregates.

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