Quantification of Fusarium culmorum in wheat and barley tissues using real-time PCR in comparison with DON content

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027006%3A_____%2F06%3A1202" target="_blank" >RIV/00027006:_____/06:1202 - isvavai.cz</a>

Result on the web

—

DOI - Digital Object Identifier

—

Result language

angličtina

Original language name

Quantification of Fusarium culmorum in wheat and barley tissues using real-time PCR in comparison with DON content

Original language 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 c

Czech name

Kvantitativní stanovení Fusarium culmorum v pletivech pšenice a ječmene pomocí real-time PCR ve srovnání s obsahem DON

Czech description

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.

Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

GJ - Diseases and animal vermin, veterinary medicine

OECD FORD branch

—

Project

<a href="/en/project/QG50076" target="_blank" >QG50076: Analysis of Fusarium head blight pathogens of wheat in the Czech Republic, specification of effects of infection on hygienic quality of grain and introduction of differential control of the disease</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2006

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Journal of Phytopathology

ISSN

0931-1785

e-ISSN

—

Volume of the periodical

154

Issue of the periodical within the volume

10

Country of publishing house

DE - GERMANY

Number of pages

9

Pages from-to

603-611

UT code for WoS article

—

EID of the result in the Scopus database

—