scholarly journals The Genome Sequence of the Tomato-Pathogenic Actinomycete Clavibacter michiganensis subsp. michiganensis NCPPB382 Reveals a Large Island Involved in Pathogenicity

2008 ◽  
Vol 190 (6) ◽  
pp. 2138-2149 ◽  
Author(s):  
Karl-Heinz Gartemann ◽  
Birte Abt ◽  
Thomas Bekel ◽  
Annette Burger ◽  
Jutta Engemann ◽  
...  
Keyword(s):  
Serine Proteases ◽  
Clavibacter Michiganensis ◽  
Reduced Sulfur Compounds ◽  
Apparent Lack ◽  
Clavibacter Michiganensis Subsp ◽  
Related Organism ◽  
Large Island ◽  
Genes Encoding ◽  
Insertion Elements ◽  
Small Stable Rna

ABSTRACT Clavibacter michiganensis subsp. michiganensis is a plant-pathogenic actinomycete that causes bacterial wilt and canker of tomato. The nucleotide sequence of the genome of strain NCPPB382 was determined. The chromosome is circular, consists of 3.298 Mb, and has a high G+C content (72.6%). Annotation revealed 3,080 putative protein-encoding sequences; only 26 pseudogenes were detected. Two rrn operons, 45 tRNAs, and three small stable RNA genes were found. The two circular plasmids, pCM1 (27.4 kbp) and pCM2 (70.0 kbp), which carry pathogenicity genes and thus are essential for virulence, have lower G+C contents (66.5 and 67.6%, respectively). In contrast to the genome of the closely related organism Clavibacter michiganensis subsp. sepedonicus, the genome of C. michiganensis subsp. michiganensis lacks complete insertion elements and transposons. The 129-kb chp/tomA region with a low G+C content near the chromosomal origin of replication was shown to be necessary for pathogenicity. This region contains numerous genes encoding proteins involved in uptake and metabolism of sugars and several serine proteases. There is evidence that single genes located in this region, especially genes encoding serine proteases, are required for efficient colonization of the host. Although C. michiganensis subsp. michiganensis grows mainly in the xylem of tomato plants, no evidence for pronounced genome reduction was found. C. michiganensis subsp. michiganensis seems to have as many transporters and regulators as typical soil-inhabiting bacteria. However, the apparent lack of a sulfate reduction pathway, which makes C. michiganensis subsp. michiganensis dependent on reduced sulfur compounds for growth, is probably the reason for the poor survival of C. michiganensis subsp. michiganensis in soil.

scholarly journals Comparative Genomic Analyses of Clavibacter michiganensis subsp. insidiosus and Pathogenicity on Medicago truncatula

2018 ◽  
Vol 108 (2) ◽  
pp. 172-185 ◽  
Author(s):  
You Lu ◽  
Carol A. Ishimaru ◽  
Jane Glazebrook ◽  
Deborah A. Samac
Keyword(s):  
Medicago Truncatula ◽  
Serine Proteases ◽  
Plant Pathogens ◽  
Blue Pigment ◽  
Comparative Genomic ◽  
Clavibacter Michiganensis ◽  
Genome Sequences ◽  
Gram Positive ◽  
Model Legume ◽  
Clavibacter Michiganensis Subsp

Clavibacter michiganensis is the most economically important gram-positive bacterial plant pathogen, with subspecies that cause serious diseases of maize, wheat, tomato, potato, and alfalfa. Much less is known about pathogenesis involving gram-positive plant pathogens than is known for gram-negative bacteria. Comparative genome analyses of C. michiganensis subspecies affecting tomato, potato, and maize have provided insights on pathogenicity. In this study, we identified strains of C. michiganensis subsp. insidiosus with contrasting pathogenicity on three accessions of the model legume Medicago truncatula. We generated complete genome sequences for two strains and compared these to a previously sequenced strain and genome sequences of four other subspecies. The three C. michiganensis subsp. insidiosus strains varied in gene content due to genome rearrangements, most likely facilitated by insertion elements, and plasmid number, which varied from one to three depending on strain. The core C. michiganensis genome consisted of 1,917 genes, with 379 genes unique to C. michiganensis subsp. insidiosus. An operon for synthesis of the extracellular blue pigment indigoidine, enzymes for pectin degradation, and an operon for inositol metabolism are among the unique features. Secreted serine proteases belonging to both the pat-1 and ppa families were present but highly diverged from those in other subspecies.

scholarly journals Characterization of the Plasmid Encoded Virulence Region pat-1 of Phytopathogenic Clavibacter michiganensis subsp. michiganensis

1997 ◽  
Vol 10 (2) ◽  
pp. 195-206 ◽  
Author(s):  
Jens Dreier ◽  
Dietmar Meletzus ◽  
Rudolf Eichenlaub
Keyword(s):  
Structural Gene ◽  
Dna Sequences ◽  
Repetitive Sequence ◽  
Transcription Initiation ◽  
Serine Proteases ◽  
Tandem Repeats ◽  
Sequence Motif ◽  
Clavibacter Michiganensis ◽  
Reading Frame ◽  
Clavibacter Michiganensis Subsp

The tomato pathogen Clavibacter michiganensis subsp. michiganensis NCPPB382, causing bacterial wilt and canker, harbors two plasmids, pCM1 (27.5 kb) and pCM2 (72 kb), carrying genes involved in virulence. The region of plasmid pCM2 encoding the pathogenicity locus pat-1 was mapped by deletion analysis and complementation studies to a 1.5-kb BglII/SmaI DNA fragment. Introduction of the pat-1 region into endophytic, plasmid-free isolates of C. michiganensis subsp. michiganensis converted these bacteria into virulent pathogens. Based on the nucleotide sequence of the pat-1 region, an open reading frame (ORF1) can be predicted, coding for a protein of 280 amino acids and 29.7 kDa with homology to serine proteases. Introduction of a frame-shift mutation in ORF1 leads to a loss of the pathogenic phenotype. Northern (RNA) hybridizations identified an 1.5-knt transcript of the pat-1 structural gene. The site of transcription initiation was mapped by primer extension and a typical -10/-35 region was located with significant homology to the consensus Escherichia coli σ70 and Bacillus subtilis σ43 promoters. Downstream of the pat-1 structural gene, a peculiar repetitive sequence motif (pat-1rep) is located, consisting of 20 direct tandem repeats preceded by a run of 14 guanosine residues. DNA sequences homologous to pat-1rep were isolated and characterized from four virulent C. michiganensis subsp. mich-iganensis strains exhibiting a high extent of structural conservation. The deletion of this repetitive sequence reduced virulence significantly but did not lead to a complete loss of the virulence phenotype.

scholarly journals Sequential Expression of Bacterial Virulence and Plant Defense Genes During Infection of Tomato with Clavibacter michiganensis subsp. michiganensis

2010 ◽  
Vol 100 (3) ◽  
pp. 252-261 ◽  
Author(s):  
L. Chalupowicz ◽  
M. Cohen-Kandli ◽  
O. Dror ◽  
R. Eichenlaub ◽  
K.-H. Gartemann ◽  
...  
Keyword(s):  
Host Defense ◽  
Serine Proteases ◽  
Bacterial Virulence ◽  
Transcriptional Analysis ◽  
Clavibacter Michiganensis ◽  
Defense Genes ◽  
Virulence Plasmids ◽  
Early Stages ◽  
Clavibacter Michiganensis Subsp ◽  
Pathogenesis Related Protein

The molecular interactions between Clavibacter michiganensis subsp. michiganensis and tomato plant were studied by following the expression of bacterial virulence and host-defense genes during early stages of infection. The C. michiganensis subsp. michiganensis genes included the plasmid-borne cellulase (celA) and the serine protease (pat-1), and the serine proteases chpC and ppaA, residing on the chp/tomA pathogenicity island (PAI). Gene expression was measured following tomato inoculation with Cmm382 (wild type), Cmm100 (lacking the plasmids pCM1 and pCM2), and Cmm27 (lacking the PAI). Transcriptional analysis revealed that celA and pat-1 were significantly induced in Cmm382 at initial 12 to 72 h, whereas chpC and ppaA were highly expressed only 96 h after inoculation. Interdependence between the expression of chromosomal and of plasmid-located genes was revealed: expression of celA and pat-1 was substantially reduced in the absence of the chp/tomA PAI, whereas chpC and ppaA expressions were reduced in the absence of the virulence plasmids. Transcription of chromosomal genes involved in cell wall degradation (i.e., pelA1, celB, xysA, and xysB), was also induced at early stages of infection. Expression of the host-defense genes, chitinase class II and pathogenesis-related protein-5 isoform was induced in the absence of the PAI at early stages of infection, suggesting that PAI-located genes are involved in suppression of tomato basal defenses.

scholarly journals Silencing of Host Basal Defense Response-Related Gene Expression Increases Susceptibility of Nicotiana benthamiana to Clavibacter michiganensis subsp. michiganensis

2011 ◽  
Vol 101 (3) ◽  
pp. 349-357 ◽  
Author(s):  
Vasudevan Balaji ◽  
Guido Sessa ◽  
Christine D. Smart
Keyword(s):  
Defense Response ◽  
Nicotiana Benthamiana ◽  
Plant Responses ◽  
Virus Induced Gene Silencing ◽  
In Planta ◽  
Clavibacter Michiganensis ◽  
Canker Disease ◽  
Clavibacter Michiganensis Subsp ◽  
Genes Encoding ◽  
Basal Defense

Clavibacter michiganensis subsp. michiganensis is an actinomycete, causing bacterial wilt and canker disease of tomato (Solanum lycopersicum). We used virus-induced gene silencing (VIGS) to identify genes playing a role in host basal defense response to C. michiganensis subsp. michiganensis infection using Nicotiana benthamiana as a model plant. A preliminary VIGS screen comprising 160 genes from tomato known to be involved in defense-related signaling identified a set of 14 genes whose suppression led to altered host–pathogen interactions. Expression of each of these genes and three additional targets was then suppressed in larger-scale VIGS experiments and the effect of silencing on development of wilt disease symptoms and bacterial growth during an N. benthamiana–C. michiganensis subsp. michiganensis compatible interaction was determined. Disease susceptibility and in planta bacterial population size were enhanced by silencing genes encoding N. benthamiana homologs of ubiquitin activating enzyme, snakin-2, extensin-like protein, divinyl ether synthase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase 2, and Pto-like kinase. The identification of genes having a role in the host basal defense-response to C. michiganensis subsp. michiganensis advances our understanding of the plant responses activated by C. michiganensis subsp. michiganensis and raises possibilities for devising novel and effective molecular strategies to control bacterial canker and wilt in tomato.

Clavibacter michiganensis subsp. sepedonicus. [Distribution map].

2017 ◽  
Author(s):  
Keyword(s):  
Solanum Tuberosum ◽  
New Brunswick ◽  
Prince Edward Island ◽  
Distribution Map ◽  
Clavibacter Michiganensis ◽  
Central Russia ◽  
Clavibacter Michiganensis Subsp ◽  
Northern Russia ◽  
New Distribution ◽  
Distribution In Europe

Abstract A new distribution map is provided for Clavibacter michiganensis subsp. sepedonicus (Spieckermann & Kotthoff) Dye & Kemp. Actinobacteria: Actinomycetales: Microbacteriaceae. Hosts: potato (Solanum tuberosum) and tomato (Solanum lycopersicum). Information is given on the geographical distribution in Europe (Austria, Belarus, Belgium, Bulgaria, Cyprus, Czech Republic, Denmark, Estonia, Finland, Germany, Greece, Crete, Hungary, Latvia, Lithuania, Netherlands, Norway, Poland, Romania, Russia, Central Russia, Northern Russia, Siberia, Slovakia, Spain, Sweden, UK and Ukraine), Asia (China, Anhui, Gansu, Hebei, Heilongjiang, Henan, Jiangsu, Ningxia, Shaanxi, Yunnan, Zhejiang, Japan, Kazakhstan, Korea Democratic People's Republic, Korea Republic, Nepal, Pakistan, Taiwan, Turkey and Uzbekistan), North America (Canada, Alberta, British Columbia, Manitoba, New Brunswick, Newfoundland, Nova Scotia, Ontario, Prince Edward Island, Quebec, Saskatchewan, Mexico, USA, Idaho, Kansas, North Dakota and Oregon) and South America (Bolivia).

Clavibacter michiganensis subsp. michiganensis strains virulence and genetic diversity. a first study in Argentina

2017 ◽  
Vol 149 (1) ◽  
pp. 35-42 ◽  
Author(s):  
Eliana Wassermann ◽  
Marcela Susana Montecchia ◽  
Olga Susana Correa ◽  
Vega Damián ◽  
Ana María Romero
Keyword(s):  
Genetic Diversity ◽  
Clavibacter Michiganensis ◽  
Clavibacter Michiganensis Subsp

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.

Characterization of three alternative splice variants associated with the Arabidopsis rhomboid protein gene At1g74130

Botany ◽  
2013 ◽  
Vol 91 (12) ◽  
pp. 840-849 ◽  
Author(s):  
Joshua Powles ◽  
Katharine Sedivy-Haley ◽  
Eric Chapman ◽  
Kenton Ko
Keyword(s):  
Alternative Splicing ◽  
Alternative Splice ◽  
Splice Variant ◽  
Serine Proteases ◽  
Transmembrane Domain ◽  
Splice Variants ◽  
Genes Encoding ◽  
Different Characteristics ◽  
Alternative Splice Variants

Rhomboid serine proteases are grouped into three main types — secretases, presenilin-like associated rhomboid-like (PARL) proteases, and “inactive” rhomboid proteins. Although the three rhomboid groups are distinct, the different types are likely to operate within the same cell or compartment, such as observed in the plastids of Arabidopsis. There are four distinct plastid rhomboid genes at play in Arabidopsis plastids, two for active types (At1g25290 and At5g25752) and two for inactive forms (At1g74130 and At1g74140). The number of working plastid rhomboids is further increased by alternative splicing, as reported for At1g25290. To understand how the plastid rhomboid system works, it is necessary to identify all rhomboid forms in play. To this end, this study was designed to examine the alternative splicing activities of At1g74130, one of the two genes encoding proteolytically “inactive” plastid rhomboids. The exon mapping and DNA sequencing results obtained here indicate the presence of three prominent alternative splice variants in the At1g74130 transcript population. The dominant splice variant, L, encodes the full-length protein. The other two splice variants, M and S, produce proteins lacking sections from the carboxyl transmembrane domain region. The splice variants M and S appear to be at levels with functional potential and appear to adjust relative to each other during development and in response to changes in the level of Tic40, a component of the plastid translocon. The splice variant proteins themselves exhibit different characteristics with respect to rhomboid protein–substrate interactions. These differences were observed in bacterial co-expression pull-down assays and in yeast mitochondrial studies. When considered together, the data suggest that the alternative splicing of At1g74130 bears functional significance in Arabidopsis and is likely to be part of a mechanism for diversifying plastid rhomboid function.

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