Clavibacter
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Clavibacter sp. strain CF11, which was isolated from soil at a tomato-planting greenhouse in Inner Mongolia, North China, has a high capability for producing cold-active cellulase at low temperatures. Here, we report the draft genome sequence of strain CF11, which comprises 2,437 protein-coding sequences and 49 RNA-coding sequences.
- Publikační typ
- časopisecké články MeSH
Detection of Clavibacter michiganensis subsp. michiganensis (Cmm), causing bacterial canker of tomato, was verified using PTA-ELISA and IFAS with PAbs of Neogen Europe Ltd. (UK), and with published and also laboratory-generated PCR primers from the Cmm tomatinase gene. The specificity of this technique was determined with 15 plant-pathogenic and 4 common, saprophytic bacteria. With IFAS, crossreactions were found for Pantoea dispersa, P. agglomerans and Rahnella aquatilis, and with PTA-ELISA for Curtobacterium flaccumfaciens, Pectobacterium atrosepticum and Dickeya sp. Cross-reactions with subspecies other than michiganensis were also found using both methods. Molecular methods were optimized by verification of annealing temperatures and times for both primers. Conditions were finally adjusted to 30 s at 65 degrees C for Dreier's and 10 s at 69 degrees C for our primer set. After this optimization, both primer pairs produced positive reaction only with Cmm. By means of PTA-ELISA and IFAS, Cmm strains were detected at a concentration up to 10(5) CFU/mL and 10(3) CFU/mL, respectively. The PCR test with bacterial cell suspensions reached a sensitivity of 10(3) CFU/mL with our designed primers and 104 CFU/mL with Dreier's primer pair.
- MeSH
- Actinomycetales MeSH
- bakteriologické techniky metody MeSH
- časové faktory MeSH
- DNA primery genetika MeSH
- ELISA metody MeSH
- financování organizované MeSH
- hodnotící studie jako téma MeSH
- hybridizace nukleových kyselin metody MeSH
- nemoci rostlin mikrobiologie MeSH
- polymerázová řetězová reakce metody MeSH
- senzitivita a specificita MeSH
- Solanum lycopersicum mikrobiologie MeSH
- teplota MeSH
- zkřížené reakce MeSH
- Publikační typ
- srovnávací studie MeSH
A multiplex real-time PCR method based on fluorescent TaqMan® probes was developed for the simultaneous detection of the tomato pathogenic bacteria Clavibacter michiganensis subsp. michiganensis, Pseudomonas syringae pv. tomato and bacterial spot-causing xanthomonads. The specificity of the multiplex assay was validated on 44 bacterial strains, including 32 target pathogen strains as well as closely related species and nontarget tomato pathogenic bacteria. The designed multiplex real-time PCR showed high sensitivity when positive amplification was observed for one pg of bacterial DNA in the cases of Clavibacter michiganensis subsp. michiganensis and Pseudomonas syringae pv. tomato bacteria and 100 pg for bacterial spot-causing xanthomonads. The reliability of the developed multiplex real-time PCR assay for in planta detection was verified by recognition of the target pathogens in 18 tomato plants artificially inoculated by each of the target bacteria and tomato samples from production greenhouses.
- MeSH
- Actinobacteria genetika izolace a purifikace fyziologie MeSH
- kvantitativní polymerázová řetězová reakce * MeSH
- prostředí kontrolované MeSH
- Pseudomonas syringae genetika izolace a purifikace fyziologie MeSH
- Solanum lycopersicum růst a vývoj mikrobiologie MeSH
- Xanthomonas genetika izolace a purifikace fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- MeSH
- DNA fingerprinting metody využití MeSH
- finanční podpora výzkumu jako téma MeSH
- grampozitivní bakterie imunologie klasifikace MeSH
- polymerázová řetězová reakce metody využití MeSH
- Publikační typ
- srovnávací studie MeSH
- Geografické názvy
- Česká republika MeSH
This work was aimed to develop a multiplex PCR assay to detect infectious agents such as Clavibacter michiganensis subsp. michiganensis, Fusarium sp, Leveillula taurica, and begomoviruses in tomato (Solanum lycopersicum) plants. Specific primer sets of each pathogen were designed based on intergenic ribosomal RNA sequences for the first three, whereas for begomoviruses, primers were designed based on conserved regions. The design also considered that the length (200-800 bp) of the PCR products was resolvable by electrophoresis; thus 296, 380, 457, and 731 bp fragments for Clavibacter, Fusarium, Leveillula, and begomoviruses, respectively, were considered. PCR conditions were optimized to amplify all the products in a single tube from genomic DNA and circumvent PCR inhibitors from infected plants. Finally, when the multiplex PCR assay was tested with tomato plants infected with any of the four pathogens, specific PCR products confirmed the presence of the pathogens. Optimized PCR multiplex allowed for the accurate and simultaneous detection of Clavibacter, Fusarium, Leveillula, and begomoviruses in infected plants or seeds from tomato.
- MeSH
- Actinomycetales genetika izolace a purifikace MeSH
- Ascomycota genetika izolace a purifikace MeSH
- Begomovirus genetika izolace a purifikace MeSH
- DNA primery genetika MeSH
- elektroforéza v polyakrylamidovém gelu MeSH
- Fusarium genetika izolace a purifikace MeSH
- multiplexová polymerázová řetězová reakce metody MeSH
- nemoci rostlin mikrobiologie virologie MeSH
- Solanum lycopersicum mikrobiologie virologie MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
... Důležité fytopatogenní bakterie 444 -- 109.1 Agrobacterium 444 -- 109.2 Burkholderia 444 -- 109.3 Clavibacter ...
Vydání první 476 stran : ilustrace, portréty ; 31 cm
Vysokoškolská učebnice, která se zaměřuje na různé druhy mikroorganismů a jejich ekologii a patogenitu.
- MeSH
- ekologie MeSH
- infekční nemoci MeSH
- mikrobiologické jevy MeSH
- mikrobiologie životního prostředí MeSH
- molekulární biologie MeSH
- Publikační typ
- monografie MeSH
- Konspekt
- Mikrobiologie
- Učební osnovy. Vyučovací předměty. Učebnice
- NLK Obory
- mikrobiologie, lékařská mikrobiologie
- NLK Publikační typ
- učebnice vysokých škol
