Welcome To Website IAS

Hot news

Independence Award

- First Rank - Second Rank - Third Rank

Labour Award

- First Rank - Second Rank -Third Rank

National Award

 - Study on food stuff for animal(2005)

 - Study on rice breeding for export and domestic consumption(2005)


- Hybrid Maize by Single Cross V2002 (2003)

- Tomato Grafting to Manage Ralstonia Disease(2005)

- Cassava variety KM140(2010)

Website links
Vietnamese calendar
Visitors summary
 Curently online :  1
 Total visitors :  4510925

Comparative Genomics to Develop a Specific Multiplex PCR Assay for Detection of Clavibacter michiganensis
Friday, 2020/05/15 | 08:30:45

Shree P. Thapa, Michael O’Leary, Marie-Agnès Jacques, Robert L. Gilbertson, and Gitta Coaker

BACTERIOLOGY J. - Published Online:31 Jan 2020 https://doi.org/10.1094/PHYTO-10-19-0405-R


Clavibacter michiganensis is a Gram-positive bacterial pathogen that proliferates in the xylem vessels of tomato, causing bacterial wilt and canker symptoms. Accurate detection is a crucial step in confirming outbreaks of bacterial canker and developing management strategies. A major problem with existing detection methods are false-positive and -negative results. Here, we report the use of comparative genomics of 37 diverse Clavibacter strains, including 21 strains sequenced in this study, to identify specific sequences that are C. michiganensis detection targets. Genome-wide phylogenic analyses revealed additional diversity within the genus Clavibacter. Pathogenic C. michiganensis strains varied in plasmid composition, highlighting the need for detection methods based on chromosomal targets. We utilized sequences of C. michiganensis-specific loci to develop a multiplex PCR-based diagnostic platform using two C. michiganensis chromosomal genes (rhuM and tomA) and an internal control amplifying both bacterial and plant DNA (16s ribosomal RNA). The multiplex PCR assay specifically detected C. michiganensis strains from a panel of 110 additional bacteria, including other Clavibacter spp. and bacterial pathogens of tomato. The assay was adapted to detect the presence of C. michiganensis in seed and tomato plant materials with high sensitivity and specificity. In conclusion, the described method represents a robust, specific tool for detection of C. michiganensis in tomato seed and infected plants.


See https://apsjournals.apsnet.org/doi/10.1094/PHYTO-10-19-0405-R

Figure 3: Clavibacter michiganensis primer design and multiplex PCR. A, Genetic architecture of flanking genes surrounding the tomA and rhuM loci. The chp/tomA cluster, including tomA, is only present in pathogenic C. michiganensisrhuM is only present in C. michiganensis tomato pathogens. Primers used to amplify each gene are shown in gray. B, Agarose gel electrophoresis of multiplex PCR assay products. Total genomic DNA was used as a template with the RhuM-F/R primers targeting the rhuM gene, the TomA-F1/R1 primers targeting the tomA gene, and the 16sR-F/R primers targeting 16S ribosomal RNA (rRNA) in either bacteria or plants. Lanes 1 to 5, endophytic Clavibacter strains (CASJ009, CFBP8017, CFBP8019, CFBP7576, and CFBP7494); lanes 6 to 39, pathogenic C. michiganensis (CFBP7311, CFBP1940, CFBP5842, CFBP7568, CFBP7488, CFBP2500, CFBP7158, CFBP7312, CFBP7316, CFBP7314, CFBP6885, CFBP7315, CFBP7589, NCPPB382, ATCC14456, ATCC10206, NZ2541, NZ1811, NZ5026, CASJ001, CASJ002, CASJ003, CASJ004, CASJ005, CASJ008, CA00001, CAYO001, AZ-28, NZ2550, 285D, 12B, 13C1, Cm4, and Cm5); lanes 40 to 43, C. sepedonicus (NZ2535, SB109, INM-1, and BRR7); lane 44, C. insidiosus (CIC266); lanes 45 and 46, C. tessellarius (CIC021 and CIC022); lanes 47 and 48, Xanthomonas euvesicatoria (RL677 and HB1); lanes 49 to 51 Pseudomonas syringae pv. tomato (PstA9, Pst18, and Pst838-8); lanes 52 to 54, Escherichia coli, ‘Candidatus Liberibacter asiaticus’ HHCA, and uninfected tomato DNA; and lane M = 1-kb DNA ladder.

Back      Print      View: 39

[ Other News ]___________________________________________________
  • Genome-wide analysis of autophagy-associated genes in foxtail millet (Setaria italica L.) and characterization of the function of SiATG8a in conferring tolerance to nitrogen starvation in rice.
  • Arabidopsis small nucleolar RNA monitors the efficient pre-rRNA processing during ribosome biogenesis
  • XA21-specific induction of stress-related genes following Xanthomonas infection of detached rice leaves.
  • Reducing the Use of Pesticides with Site-Specific Application: The Chemical Control of Rhizoctonia solani as a Case of Study for the Management of Soil-Borne Diseases
  • OsJRL, a rice jacalin-related mannose-binding lectin gene, enhances Escherichia coli viability under high-salinity stress and improves salinity tolerance of rice.
  • Production of lipopeptide biosurfactants by Bacillus atrophaeus 5-2a and their potential use in microbial enhanced oil recovery.
  • GhABF2, a bZIP transcription factor, confers drought and salinity tolerance in cotton (Gossypium hirsutum L.).
  • Resilience of cassava (Manihot esculenta Crantz) to salinity: implications for food security in low-lying regions.
  • Cellulose synthase complexes act in a concerted fashion to synthesize highly aggregated cellulose in secondary cell walls of plants
  • No adverse effects of transgenic maize on population dynamics of endophytic Bacillus subtilis strain B916-gfp
  • Identification and expression analysis of OsLPR family revealed the potential roles of OsLPR3 and 5 in maintaining phosphate homeostasis in rice
  • Functional analysis of molecular interactions in synthetic auxin response circuits
  • Titanium dioxide nanoparticles strongly impact soil microbial function by affecting archaeal nitrifiers.
  • Inducible Expression of the De-Novo Designed Antimicrobial Peptide SP1-1 in Tomato Confers Resistance to Xanthomonas campestris pv. vesicatoria.
  • Toward combined delignification and saccharification of wheat straw by a laccase-containing designer cellulosome
  • SNP-based discovery of salinity-tolerant QTLs in a bi-parental population of rice (Oryza sativa)
  • Pinpointing genes underlying the quantitative trait loci for root-knot nematode resistance in palaeopolyploid soybean by whole genome resequencing.
  • Transcriptome- Assisted Label-Free Quantitative Proteomics Analysis Reveals Novel Insights into Piper nigrum -Phytophthora capsici Phytopathosystem.
  • Brassinosteroids participate in the control of basal and acquired freezing tolerance of plants
  • Rapid hyperosmotic-induced Ca2+ responses in Arabidopsis thaliana exhibit sensory potentiation and involvement of plastidial KEA transporters
Designed & Powered by WEBSO CO.,LTD