Real-time PCR for quantification of toxigenic Fusarium species in barley and malt

Tuija Sarlin (Corresponding Author), Tapani Yli-Mattila, Marika Jestoi, Aldo Rizzo, Sari Paavanen-Huhtala, Auli Haikara

Research output: Contribution to journalArticleScientificpeer-review

58 Citations (Scopus)

Abstract

A real-time PCR technique was applied for the quantification of trichothecene-producing Fusarium species (TMTRI assay) as well as the highly toxigenic Fusarium graminearum (TMFg12 assay) present in barley grain and malt. PCR results were compared to the amounts of trichothecenes detected in the samples to find out if the PCR assays can be used for trichothecene screening instead of expensive and laborious chemical analyses. DNA was extracted from ground kernels using a commercial DNA extraction kit and analysed in a LightCycler® system using specific primers and fluorogenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium DNA and F. graminearum DNA present in barley grain and malt samples, respectively. Both TaqMan assays were considered to be sensitive and reproducible. Linearity of the assays was 4–5 log units when pure Fusarium DNAs were tested. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay could be used for estimation of the deoxynivalenol (DON) content in barley grain. Furthermore, the TMFg12 assay for F. graminearum gave a good estimation of the DON content in north American barley and malt samples, whilst the correlation was poor among Finnish samples. DON content and the level of F. graminearum DNA were found to be naturally low in Finnish barleys.
Original languageEnglish
Pages (from-to)371-380
Number of pages10
JournalEuropean Journal of Plant Pathology
Volume114
Issue number4
DOIs
Publication statusPublished - 2006
MoE publication typeA1 Journal article-refereed

Fingerprint

malt
Fusarium
quantitative polymerase chain reaction
barley
trichothecenes
assays
Fusarium graminearum
DNA
deoxynivalenol
sampling
probes (equipment)
screening
seeds

Keywords

  • Gibberella
  • trichothecenes
  • TaqMan

Cite this

Sarlin, Tuija ; Yli-Mattila, Tapani ; Jestoi, Marika ; Rizzo, Aldo ; Paavanen-Huhtala, Sari ; Haikara, Auli. / Real-time PCR for quantification of toxigenic Fusarium species in barley and malt. In: European Journal of Plant Pathology. 2006 ; Vol. 114, No. 4. pp. 371-380.
@article{4310a0a6a7724d5a9d9462d043c3a402,
title = "Real-time PCR for quantification of toxigenic Fusarium species in barley and malt",
abstract = "A real-time PCR technique was applied for the quantification of trichothecene-producing Fusarium species (TMTRI assay) as well as the highly toxigenic Fusarium graminearum (TMFg12 assay) present in barley grain and malt. PCR results were compared to the amounts of trichothecenes detected in the samples to find out if the PCR assays can be used for trichothecene screening instead of expensive and laborious chemical analyses. DNA was extracted from ground kernels using a commercial DNA extraction kit and analysed in a LightCycler{\circledR} system using specific primers and fluorogenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium DNA and F. graminearum DNA present in barley grain and malt samples, respectively. Both TaqMan assays were considered to be sensitive and reproducible. Linearity of the assays was 4–5 log units when pure Fusarium DNAs were tested. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay could be used for estimation of the deoxynivalenol (DON) content in barley grain. Furthermore, the TMFg12 assay for F. graminearum gave a good estimation of the DON content in north American barley and malt samples, whilst the correlation was poor among Finnish samples. DON content and the level of F. graminearum DNA were found to be naturally low in Finnish barleys.",
keywords = "Gibberella, trichothecenes, TaqMan",
author = "Tuija Sarlin and Tapani Yli-Mattila and Marika Jestoi and Aldo Rizzo and Sari Paavanen-Huhtala and Auli Haikara",
year = "2006",
doi = "10.1007/s10658-006-0001-9",
language = "English",
volume = "114",
pages = "371--380",
journal = "European Journal of Plant Pathology",
issn = "0929-1873",
publisher = "Springer",
number = "4",

}

Sarlin, T, Yli-Mattila, T, Jestoi, M, Rizzo, A, Paavanen-Huhtala, S & Haikara, A 2006, 'Real-time PCR for quantification of toxigenic Fusarium species in barley and malt', European Journal of Plant Pathology, vol. 114, no. 4, pp. 371-380. https://doi.org/10.1007/s10658-006-0001-9

Real-time PCR for quantification of toxigenic Fusarium species in barley and malt. / Sarlin, Tuija (Corresponding Author); Yli-Mattila, Tapani; Jestoi, Marika; Rizzo, Aldo; Paavanen-Huhtala, Sari; Haikara, Auli.

In: European Journal of Plant Pathology, Vol. 114, No. 4, 2006, p. 371-380.

Research output: Contribution to journalArticleScientificpeer-review

TY - JOUR

T1 - Real-time PCR for quantification of toxigenic Fusarium species in barley and malt

AU - Sarlin, Tuija

AU - Yli-Mattila, Tapani

AU - Jestoi, Marika

AU - Rizzo, Aldo

AU - Paavanen-Huhtala, Sari

AU - Haikara, Auli

PY - 2006

Y1 - 2006

N2 - A real-time PCR technique was applied for the quantification of trichothecene-producing Fusarium species (TMTRI assay) as well as the highly toxigenic Fusarium graminearum (TMFg12 assay) present in barley grain and malt. PCR results were compared to the amounts of trichothecenes detected in the samples to find out if the PCR assays can be used for trichothecene screening instead of expensive and laborious chemical analyses. DNA was extracted from ground kernels using a commercial DNA extraction kit and analysed in a LightCycler® system using specific primers and fluorogenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium DNA and F. graminearum DNA present in barley grain and malt samples, respectively. Both TaqMan assays were considered to be sensitive and reproducible. Linearity of the assays was 4–5 log units when pure Fusarium DNAs were tested. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay could be used for estimation of the deoxynivalenol (DON) content in barley grain. Furthermore, the TMFg12 assay for F. graminearum gave a good estimation of the DON content in north American barley and malt samples, whilst the correlation was poor among Finnish samples. DON content and the level of F. graminearum DNA were found to be naturally low in Finnish barleys.

AB - A real-time PCR technique was applied for the quantification of trichothecene-producing Fusarium species (TMTRI assay) as well as the highly toxigenic Fusarium graminearum (TMFg12 assay) present in barley grain and malt. PCR results were compared to the amounts of trichothecenes detected in the samples to find out if the PCR assays can be used for trichothecene screening instead of expensive and laborious chemical analyses. DNA was extracted from ground kernels using a commercial DNA extraction kit and analysed in a LightCycler® system using specific primers and fluorogenic TaqMan probes. Both naturally and artificially contaminated grains were analysed. The TMTRI assay and the TMFg12 assay enabled the quantification of trichothecene-producing Fusarium DNA and F. graminearum DNA present in barley grain and malt samples, respectively. Both TaqMan assays were considered to be sensitive and reproducible. Linearity of the assays was 4–5 log units when pure Fusarium DNAs were tested. The amount of Fusarium DNA analysed with the TMTRI-trichothecene assay could be used for estimation of the deoxynivalenol (DON) content in barley grain. Furthermore, the TMFg12 assay for F. graminearum gave a good estimation of the DON content in north American barley and malt samples, whilst the correlation was poor among Finnish samples. DON content and the level of F. graminearum DNA were found to be naturally low in Finnish barleys.

KW - Gibberella

KW - trichothecenes

KW - TaqMan

U2 - 10.1007/s10658-006-0001-9

DO - 10.1007/s10658-006-0001-9

M3 - Article

VL - 114

SP - 371

EP - 380

JO - European Journal of Plant Pathology

JF - European Journal of Plant Pathology

SN - 0929-1873

IS - 4

ER -