Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum Clade

Fusarium graminearum clade pathogens cause Fusarium head blight (FHB) or scab of wheat and other small cereal grains, producing different kinds of trichothecene mycotoxins that are detrimental to human and domestic animals. Type B trichothecene mycotoxins such as deoxynivalenol, 3-acetyldeoxynivalen...

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Main Authors: Jian-Hua Wang, Bo Qu, Jing-Bo Zhang, Tao Huang, Yu-Cai Liao, Fang-Fang Chen, He-Ping Li
Format: Article
Language:English
Published: MDPI AG 2008-12-01
Series:International Journal of Molecular Sciences
Subjects:
NIV
Online Access:http://www.mdpi.com/1422-0067/9/12/2495/
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spelling doaj-9417b0c647d64be8a4ab0093ebdf64c32020-11-25T02:09:29ZengMDPI AGInternational Journal of Molecular Sciences1422-00672008-12-019122495250410.3390/ijms9122495Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum CladeJian-Hua WangBo QuJing-Bo ZhangTao HuangYu-Cai LiaoFang-Fang ChenHe-Ping LiFusarium graminearum clade pathogens cause Fusarium head blight (FHB) or scab of wheat and other small cereal grains, producing different kinds of trichothecene mycotoxins that are detrimental to human and domestic animals. Type B trichothecene mycotoxins such as deoxynivalenol, 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON) and nivalenol (NIV) are the principal Fusarium mycotoxins reported in China, as well as in other countries. A genomic polymerase chain reaction (PCR) to predict chemotypes was developed based on the structural gene sequences of Tri13 genes involved in trichothecene mycotoxin biosynthesis pathways. A single pair of primers derived from the Tri13 genes detected a 583 bp fragment from 15-AcDON-chemotypes, a 644 bp fragment from 3-AcDON-chemotypes and an 859 bp fragment from NIV-producing strains. Fusarium strains from China, Nepal, USA and Europe were identified by this method, revealing their mycotoxin chemotypes identical to that obtained by chemical analyses of HPLC or GC/MS and other PCR assays. The mycotoxin chemotype-specific fragments were amplified from a highly variable region located in Tri13 genes with three deletions for 15-AcDON-chemotypes, two deletions for 3-AcDON-chemotypes and no deletion for NIV-producers. This PCR assay generated a single amplicon and thus should be more reliable than other PCR-based assays that showed the absence or presence of a PCR fragment since these assays may generate false-negative results. The results with strains from several different countries as well as from different hosts further indicated that this method should be globally applicable. This is a rapid, reliable and cost-effective method for the identification of type B trichothecene mycotoxin chemotypes in Fusarium species and food safety controls.http://www.mdpi.com/1422-0067/9/12/2495/Fusarium graminearum cladetrichothecenemycotoxin chemotype3- AcDON15-AcDONNIV
collection DOAJ
language English
format Article
sources DOAJ
author Jian-Hua Wang
Bo Qu
Jing-Bo Zhang
Tao Huang
Yu-Cai Liao
Fang-Fang Chen
He-Ping Li
spellingShingle Jian-Hua Wang
Bo Qu
Jing-Bo Zhang
Tao Huang
Yu-Cai Liao
Fang-Fang Chen
He-Ping Li
Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum Clade
International Journal of Molecular Sciences
Fusarium graminearum clade
trichothecene
mycotoxin chemotype
3- AcDON
15-AcDON
NIV
author_facet Jian-Hua Wang
Bo Qu
Jing-Bo Zhang
Tao Huang
Yu-Cai Liao
Fang-Fang Chen
He-Ping Li
author_sort Jian-Hua Wang
title Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum Clade
title_short Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum Clade
title_full Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum Clade
title_fullStr Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum Clade
title_full_unstemmed Development of a Generic PCR Detection of 3-Acetyldeoxynivalenol-, 15-Acetyldeoxynivalenol- and Nivalenol-Chemotypes of Fusarium graminearum Clade
title_sort development of a generic pcr detection of 3-acetyldeoxynivalenol-, 15-acetyldeoxynivalenol- and nivalenol-chemotypes of fusarium graminearum clade
publisher MDPI AG
series International Journal of Molecular Sciences
issn 1422-0067
publishDate 2008-12-01
description Fusarium graminearum clade pathogens cause Fusarium head blight (FHB) or scab of wheat and other small cereal grains, producing different kinds of trichothecene mycotoxins that are detrimental to human and domestic animals. Type B trichothecene mycotoxins such as deoxynivalenol, 3-acetyldeoxynivalenol (3-AcDON), 15-acetyldeoxynivalenol (15-AcDON) and nivalenol (NIV) are the principal Fusarium mycotoxins reported in China, as well as in other countries. A genomic polymerase chain reaction (PCR) to predict chemotypes was developed based on the structural gene sequences of Tri13 genes involved in trichothecene mycotoxin biosynthesis pathways. A single pair of primers derived from the Tri13 genes detected a 583 bp fragment from 15-AcDON-chemotypes, a 644 bp fragment from 3-AcDON-chemotypes and an 859 bp fragment from NIV-producing strains. Fusarium strains from China, Nepal, USA and Europe were identified by this method, revealing their mycotoxin chemotypes identical to that obtained by chemical analyses of HPLC or GC/MS and other PCR assays. The mycotoxin chemotype-specific fragments were amplified from a highly variable region located in Tri13 genes with three deletions for 15-AcDON-chemotypes, two deletions for 3-AcDON-chemotypes and no deletion for NIV-producers. This PCR assay generated a single amplicon and thus should be more reliable than other PCR-based assays that showed the absence or presence of a PCR fragment since these assays may generate false-negative results. The results with strains from several different countries as well as from different hosts further indicated that this method should be globally applicable. This is a rapid, reliable and cost-effective method for the identification of type B trichothecene mycotoxin chemotypes in Fusarium species and food safety controls.
topic Fusarium graminearum clade
trichothecene
mycotoxin chemotype
3- AcDON
15-AcDON
NIV
url http://www.mdpi.com/1422-0067/9/12/2495/
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