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Major Changes in Fusarium spp. in Wheat in the Netherlands
Authors:Cees Waalwijk  Pieter Kastelein  Ineke de Vries  Zoltan Kerényi  Theo van der Lee  Thamara Hesselink  Jürgen Köhl  Gert Kema
Institution:(1) Business Units Biointeractions and Plant Health, Plant Research International BV, Droevendaalsesteeg 1, P.O. Box 16, 6700 AA Wageningen, The Netherlands;(2) Business Units Crop and Production Ecology, Plant Research International BV, Droevendaalsesteeg 1, P.O. Box 16, 6700 AA Wageningen, The Netherlands;(3) Business Units Agricultural Biotechnology Center, Plant Research International BV, Gödöllo, Hungary
Abstract:The re-emergence of fusarium head blight throughout the world and especially in Western Europe prompted a survey of the situation in the Netherlands. To allow for a high throughput screening of large numbers of samples, a diagnostic PCR method was developed to detect the most common species of Fusarium occurring on wheat. Seven primer pairs were tested for their ability to identify isolates of Fusarium avenaceum, F. culmorum, F. graminearum, F. poae, F. proliferatum and Microdochium nivale var. majus and M. nivale var. nivale. Each primer pair only generated a PCR product with the corresponding Fusarium species and all PCR fragments had different molecular sizes. This allowed the generation of these amplicons using a mixture of all seven primer pairs. The robustness of this multiplex PCR encouraged us to screen a large series of isolates collected in 2000 and 2001. In both years 40 fields were sampled leading to a collection of 209 isolates from 2000 and 145 isolates from 2001. The results of the multiplex PCR demonstrated that F. graminearum was the most abundant species in the Fusarium complex on wheat in both years. This is in sharp contrast to reports from the 1980s and early 1990s, which found F. culmorum as the predominant species. Primers derived from the tri7 and tri13 genes, which are implicated in the acetylation and oxygenation of the C-4 atom of the backbone of the trichothecene molecule, were used to discriminate between deoxynivalenol and nivalenol (NIV) producers. The populations of F. culmorum and F. graminearum both showed a slight increase in NIV-producers in 2001.
Keywords:fusarium head blight  multiplex PCR  population diversity  molecular identification  mycotoxin
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