Specific Detection and Identification of Fusarium graminearum Sensu Stricto Using a PCR-RFLP Tool and Specific Primers Targeting the Translational Elongation Factor 1α Gene

Abstract

Fusarium graminearum is a toxigenic plant pathogen that causes Fusarium head blight (FHB) disease on cereal crops. It has recently shown to have cross-pathogenicity on noncereals (i.e., Fusarium root rot [FRR] on soybean) in Canada and elsewhere. Specific detection and differentiation of this potent toxigenic, trichothecene-producing pathogen among other closely related species is extremely important for disease control and mycotoxin monitoring. Here, we designed a PCR restriction fragment length polymorphism protocol based on the DNA sequence of the translational elongation factor 1α (TEF1α) gene. A unique restriction site to the enzyme HpaII is only found in F. graminearum sensu stricto strains among different Fusarium strains in the F. graminearum species complex (FGSC) and other Fusarium spp. associated with FHB in cereals and FRR in soybean. Partial amplification of the TEF1α gene with newly designed primers mh1/mh2 generated a 459-bp PCR fragment. Restriction digestion of the generated fragments with the HpaII enzyme generated a unique restriction pattern that can rapidly and accurately differentiate F. graminearum sensu stricto among all other Fusarium spp. A primer pair (FgssF/FgssR) specific to F. graminearum sensu stricto also was designed and can distinguish F. graminearum sensu stricto from all other Fusarium spp. in the FGSC and other closely related Fusarium spp. involved in FHB and FRR. This finding will be very useful for the specific detection of F. graminearum sensu stricto for diagnostic purposes as well as for the accurate detection of this pathogen in breeding and other research purposes.

Phenotypic and molecular characterization of the resistance to azoxystrobin and pyraclostrobin inFusarium graminearumpopulations from Brazil

“…The unidentified strains from PR state were assayed using the Fg16‐F/R primers, which allow differentiation between F . graminearum (<500 bp) and Fusarium meridionale (>500 bp) according to the band product size (Hafez et al, 2020; Nicholson et al, 1998). Primer sequences and product size can be found in Table 1.…”

Effect of Plasma-Activated Water Bubbles on Fusarium graminearum, Deoxynivalenol, and Germination of Naturally Infected Barley during Steeping

“…Tri5 , Tri6 , EF1-A and β-Actin genes were used for F. graminearum detection and relative quantification based on qPCR [ 46 , 47 , 48 , 49 ], in order to confirm the results obtained with the plating technique, which indicated that the PAW bubble treatment did not have any effect on F. graminearum infection or growth. DON biosynthesis is governed by fifteen genes that are present over three chromosomes [ 50 ] and an acidic pH of environment encourages DON biosynthesis.…”

“…Four F. graminearum -specific genes were targeted. The full sequence of the genes, including Trichothecene biosynthesis transcription regulator ( Tri6 , GenBank: AB017495.1), Terpene cyclase trichodiene synthase ( Tri5 ; GenBank: KJ677974.1) [ 46 ], Elongation factor 1-alpha ( EF1 A ; GenBank: KX084040.1) [ 47 ], and Actin (NCBI Reference Sequence: XM_011328784.1) [ 48 ] were retrieved from the NCBI sequence database. Gene-specific primers ( Table 6 ) were designed using Primer Express 3.0.1 (Applied Biosystems, Mississauga, ON, Canada).…”

An Efficient Strategy Combining Immunoassays and Molecular Identification for the Investigation of Fusarium Infections in Ear Rot of Maize in Guizhou Province, China

“…In addition, there are no orthologous copies of TEF-1 α in this genus, making this locus a better candidate for distinguishing phylogenetic relationships ( Amarasinghe et al, 2019 ). Therefore, classification and identification of Fusarium species based on TEF-1 α gene sequences has become the most common method ( Mirete et al, 2004 ; Hafez et al, 2020 ). Geiser et al (2004) established a Fusarium database based on the partial sequence of the TEF-1 α gene, which allows researchers to easily identify species under Fusarium spp.…”