Abstract
The occurrence resistance to methyl benzimidazole carbamates (MBC)-fungicides in the Fusarium graminearum species complex (FGSC) is becoming a serious problem in the control of Fusarium head blight in China. The resistance is caused by point mutations in the β2-tubulingene. So far, five resistant genotypes (F167Y, E198Q, E198L, E198K and F200Y) have been reported in the field. To establish a high-throughput method for rapid detection of all the five mutations simultaneously, an efficient single-nucleotide-polymorphism-based genotyping method was developed based on the Luminex xMAP system. One pair of amplification primers and five allele specific primer extension probes were designed and optimized to specially distinguish the different genotypes within one single reaction. This method has good extensibility and can be combined with previous reported probes to form a highly integrated tool for species, trichothecene chemotype and MBC resistance detection. Using this method, carbendazim resistant FGSC isolates from Jiangsu, Anhui and Sichuan Province in China were identified. High and moderate frequencies of resistance were observed in Jiangsu and Anhui Province, respectively. Carbendazim resistance in F. asiaticum is only observed in the 3ADON genotype. Overall, our method proved to be useful for early detection of MBC resistance in the field and the result aids in the choice of fungicide type.
Highlights
Increased resistance to the most commonly used fungicide, carbendazim, is escalating Fusarium head blight problems in China
We developed a fast SNPs-based genotyping assay for the detection of all the five known methyl benzimidazole carbamates (MBC) resistant genotypes of Fusarium graminearum species complex (FGSC), which can be used complementary to the time consuming antifungal resistance tests based on plating and cultivation
F. asiaticum isolate js205, which is highly resistant to MBC because of a point mutation at codon 167 (Phe to Tyr, F167Y) in the Tub2 gene, F. asiaticum isolate js161, which is highly resistant to MBC because of a point mutation at codon 200 (Phe to Tyr, F200Y) in the Tub2 gene, F. asiaticum isolate js801, which is moderately resistant to MBC because of a point mutation at codon 198 (Glu to Gln, E198Q) in the Tub2 gene and F. asiaticum isolate js166, which is sensitive to MBC, were used as reference in this study (Table 1)
Summary
Increased resistance to the most commonly used fungicide, carbendazim, is escalating Fusarium head blight problems in China. Fusarium head blight (FHB) or head scab is a devastating disease of wheat (Triticum aestivum L) and barley (Hordeum vulgare L) in China (Zhang et al, 2012; Zhang et al, 2010) and many other regions of the world (Suga et al, 2008; Talas, Parzies & Miedaner, 2011). Due to the changes in climatic conditions and in agricultural practices, FHB outbreaks have occurred more frequently in China, leading to a significant yield loss in more than 5 million ha (approximately 20%) of wheat grown each year from 2008 to 2015
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