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Statements

Subject Item
n2:RIV%2F00027006%3A_____%2F06%3A1202%21RIV08-MZE-00027006
rdf:type
n12:Vysledek skos:Concept
dcterms:description
A real-time polymerase chain reaction (PCR) assay was developed for the specific detection of Fusarium culmorum in infected seeds. Primers and TaqMan minor groove binder probe were derived from the sequence of a F. culmorum specific PCR product. The specificity of the assay was confirmed by test in seven Fusarium species and 21 non-Fusarium fungal species. With serial dilutions of purified genomic DNA from F. culmorum isolate B as a template, the detection limit of the assay was found to be 0,9 pg of fungal genomic DNA per reaction. A significant correlation (r2=0,982) and collinearity was found between DNA concentration and Ct values of real-time PCR assay with serial diluted DNAs extracted from three seed samples with different DON content. Eight barley and nine wheat varieties infected by F. culmorum isolate B were evaluated in 1 (barley samples) and in 4 (wheat samples) years. The results of real-time PCR analysis and enzyme-linked immunosorbent assay testing for DON content were compared and a s PCR reakce v reálném čase byla použita pro detekci patogenní houby Fusarium culmorum v pletivech pšenice a ječmene. Byla vyvinuta specifická sonda a primery pro danou reakci. Specificita reakce byla testována na 7 druzích hub rodu Fusarium a 21 druzích jiných druhů patogenních hub. Jako standard byla použita seriově naředěná DNA extrahovaná z mycelia izolátu B Fusarium culmorum. Na základě ředicí řady byla vytvořena kalibrační křivka. Odečtem hodnot Ct byla stanovana koncentrace F. culmorum v daném vzorku. Byly porovnány hodnoty Ct s hodnotami DON stanovenými metodou ELISA. Metoda real-time PCR se ukázala jako vysoce specifická a citlivá pro stanovení obsahu patogena v pletivech hostitele a může být použita ve fytotatologických studiích a v praxi. A real-time polymerase chain reaction (PCR) assay was developed for the specific detection of Fusarium culmorum in infected seeds. Primers and TaqMan minor groove binder probe were derived from the sequence of a F. culmorum specific PCR product. The specificity of the assay was confirmed by test in seven Fusarium species and 21 non-Fusarium fungal species. With serial dilutions of purified genomic DNA from F. culmorum isolate B as a template, the detection limit of the assay was found to be 0,9 pg of fungal genomic DNA per reaction. A significant correlation (r2=0,982) and collinearity was found between DNA concentration and Ct values of real-time PCR assay with serial diluted DNAs extracted from three seed samples with different DON content. Eight barley and nine wheat varieties infected by F. culmorum isolate B were evaluated in 1 (barley samples) and in 4 (wheat samples) years. The results of real-time PCR analysis and enzyme-linked immunosorbent assay testing for DON content were compared and a s
dcterms:title
Quantification of Fusarium culmorum in wheat and barley tissues using real-time PCR in comparison with DON content Kvantitativní stanovení Fusarium culmorum v pletivech pšenice a ječmene pomocí real-time PCR ve srovnání s obsahem DON Quantification of Fusarium culmorum in wheat and barley tissues using real-time PCR in comparison with DON content
skos:prefLabel
Kvantitativní stanovení Fusarium culmorum v pletivech pšenice a ječmene pomocí real-time PCR ve srovnání s obsahem DON Quantification of Fusarium culmorum in wheat and barley tissues using real-time PCR in comparison with DON content Quantification of Fusarium culmorum in wheat and barley tissues using real-time PCR in comparison with DON content
skos:notation
RIV/00027006:_____/06:1202!RIV08-MZE-00027006
n3:strany
603;611
n3:aktivita
n10:P
n3:aktivity
P(QG50076)
n3:cisloPeriodika
10
n3:dodaniDat
n15:2008
n3:domaciTvurceVysledku
n9:7746393 n9:9285555 n9:2129485 n9:9505024 n9:5482135 n9:1933795
n3:druhVysledku
n17:J
n3:duvernostUdaju
n16:S
n3:entitaPredkladatele
n6:predkladatel
n3:idSjednocenehoVysledku
496184
n3:idVysledku
RIV/00027006:_____/06:1202
n3:jazykVysledku
n13:eng
n3:klicovaSlova
barley; wheat; DON; Fusarium culmorum; real-time PCR
n3:klicoveSlovo
n4:barley n4:Fusarium%20culmorum n4:real-time%20PCR n4:DON n4:wheat
n3:kodStatuVydavatele
DE - Spolková republika Německo
n3:kontrolniKodProRIV
[D0AEF27547E0]
n3:nazevZdroje
Journal of Phytopathology
n3:obor
n11:GJ
n3:pocetDomacichTvurcuVysledku
6
n3:pocetTvurcuVysledku
6
n3:projekt
n14:QG50076
n3:rokUplatneniVysledku
n15:2006
n3:svazekPeriodika
154
n3:tvurceVysledku
Ovesná, Jaroslava Šíp, Václav Leišová, Leona Kučera, Ladislav Sýkorová, Světlana Chrpová, Jana
s:issn
0931-1785
s:numberOfPages
9