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

Plant Disease ◽  
2020 ◽  
Vol 104 (4) ◽  
pp. 1076-1086
Author(s):  
Mohamed Hafez ◽  
Ahmed Abdelmagid ◽  
Lorne R. Adam ◽  
Fouad Daayf
Keyword(s):  
Fusarium Graminearum ◽  
Elongation Factor ◽  
Sensu Stricto ◽  
Restriction Pattern ◽  
Head Blight ◽  
Specific Detection ◽  
Fusarium Spp ◽  
Elongation Factor 1Α ◽  
Detection And Identification ◽  
Unique Restriction

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.

scholarly journals Trichothecene Genotype of Fusarium graminearum Isolates from Soybean (Glycine max) Seedling and Root Diseases in the United States

Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 1012-1012 ◽  
Author(s):  
M. L. Ellis ◽  
G. P. Munkvold
Keyword(s):  
United States ◽  
North America ◽  
Fusarium Graminearum ◽  
Root Rot ◽  
Elongation Factor ◽  
Host Shift ◽  
The United States ◽  
Sensu Stricto ◽  
Head Blight ◽  
The Third

Fusarium graminearum is an economically important pathogen that causes Fusarium head blight of wheat, barley, and oat, and Gibberella ear and stalk rot of maize. More recently, F. graminearum was reported as a soybean seedling and root pathogen in North America (1,5), causing seed decay, damping-off, and brown to reddish-brown root rot symptoms. Type B trichothecene mycotoxins are commonly produced by F. graminearum, which can be categorized into three trichothecene genotypes; those that produce 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), or nivalenol (NIV). The 15-ADON genotype is dominant in populations from small grains and maize in North America (4), but the 3-ADON genotype has recently been found (4). F. graminearum was known as a pathogen of wheat and maize in North America for over a century before it was reported as a soybean pathogen. Therefore, we hypothesized that recent reports on soybean could be associated with the appearance of the 3-ADON genotype. The objective of this research was to determine the trichothecene genotype of F. graminearum isolates from soybean in the United States. Thirty-eight isolates from soybean were evaluated. Twenty-seven isolates came from a 3-year survey for Fusarium root rot from 2007 to 2009 in Iowa. Other isolates (Ahmad Fakhoury, Southern Illinois University, Carbondale) were collected from soybean seedlings during a multi-state survey in 2012, and included three isolates from Illinois, three from Indiana, and five from Nebraska. Species identification and lineage of F. graminearum were confirmed by sequencing the translation elongation factor gene (EF1-α) using EF-1H and EF-2T primers. A maximum likelihood analysis of the EF1-α, including voucher strains from nine lineages of F. graminearum (2), placed all 38 isolates into lineage 7, F. graminearum sensu stricto (representative GenBank accessions KJ415349 to KJ415352). To determine the trichothecene genotype of each isolate we used three multiplex PCR assays. The first two assays targeted a portion of trichothecene biosynthesis genes Tri3 and Tri12 (4), while the third assay targeted portions of the Tri3, Tri5, and Tri7 genes (3). The PCR for the first two assays was conducted as described by Ward et al. (4) using four sets of primers: 3CON, 3NA, 3D15A, and 3D3A; and 12CON, 12NF, 12-15F, and 12-3F for the Tri3 and Tri12 genes, respectively. The PCR for the third assay was conducted as described by Quarta et al. (3) using the following primers: Tri3F971, Tri3F1325, Tri3R1679, Tri7F340, Tri7R965, 3551H, and 4056H. The amplification products were analyzed by gel electrophoresis. All 38 isolates produced amplicons consistent with the 15-ADON genotype; ~610 and 670 bp for the Tri3 and Tri12 genes, respectively (4), and two amplicons of ~708 and 525 bp for the Tri3/Tri5 genes (3). Our results indicated that the dominant trichothecene genotype among isolates of F. graminearum from soybean is 15-ADON, and the introduction of 3-ADON isolates does not explain the recent host shift of F. graminearum to soybean in North America. To our knowledge, this is the first assessment of trichothecene genotypes in F. graminearum populations from soybean from the United States. References: (1) K. E. Broders et al. Plant Dis. 91:1155, 2007. (2) K. O'Donnell et al. Fungal Gen. Biol. 41:600, 2004. (3) A. Quarta et al. FEMS Microbiol. Lett. 259:7, 2006. (4) T. D. Ward et al. Fungal Gen. Biol. 45:473, 2008. (5) A. G. Zue et al. Can. J. Plant Pathol. 29:35, 2007.

scholarly journals Trichothecene Genotypes of Fusarium graminearum Populations Isolated from Winter Wheat Crops in Serbia

Toxins ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 460 ◽  
Author(s):  
Vesna Krnjaja ◽  
Slavica Stanković ◽  
Ana Obradović ◽  
Tanja Petrović ◽  
Violeta Mandić ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Gene Clusters ◽  
Sensu Stricto ◽  
Molecular Techniques ◽  
Weather Data ◽  
Morphological Observation ◽  
Specific Primer ◽  
Head Blight ◽  
As Species ◽  
Species Specific

Fusarium graminearum as the main causal agent of Fusarium head blight (FHB) and its ability to produce trichothecenes was investigated by molecular techniques. A total of 37 strains isolated from the wheat, harvested in Serbia in 2005, 2008 and 2015, and previously designated by morphological observation as F. graminearum, were used for trichothecene genotypes characterization. The strains were identified using the species-specific primer set FG16R/FG16F while genotypic characterization was done using specific TRI13 and TRI3 sequences of the trichothecene gene clusters. The PCR assays identified all strains as species of F. graminearum sensu stricto with the DON/15-ADON genotype. The quantification of the mycotoxin (DON) was performed using the biochemical assay. The high levels of DON (>20,000 µg kg−1) were recorded in all of the strains from 2005, four strains from 2008 and two strains from 2015. Weather data of the investigated seasons, showed that the optimal temperature, frequent rains and high relative humidity (RH) was very favourable for the development of F. graminearum, affecting the DON biosynthesis.

scholarly journals The Effect of Fusarium Graminearum Inoculation on Winter Wheat at ARDS Turda,

2012 ◽  
Vol 69 (1) ◽  
Author(s):  
Rozalia KADAR ◽  
Amin Said SARDAR Amin Said SARDAR
Keyword(s):  
Winter Wheat ◽  
Fusarium Graminearum ◽  
Head Blight ◽  
Fusarium Spp ◽  
Flowering Stage ◽  
Stage Number ◽  
Precise Data

Fusarium  Head Blight (FHB), caused by Fusarium spp., has become one of the most destructive diseases in the world’s wheat growing areas, especially in humid and semihumid regions. More precise data relating the effects of FHB on yield have been obtained using inoculated trials. The situation is totally changed by inoculation with Fusarium. Because this disease affected wheat in flowering stage, number of grain/spike is strongly diminished. It can be observed the behavior of Turda 95 and Dumbrava varieties created at ARDS Turda which lose less number of grain than other cultivars.

Biological Control of Fusarium Head Blight of What Caused by Fusarium Graminearum and Fusarium spp. of Section Sporotrichiella

1997 ◽  
Vol 25 (3) ◽  
pp. 763-764
Author(s):  
G. V. Kalko ◽  
L. A. Nasarovskaya ◽  
I. I. Novikova ◽  
I. V. Boikova ◽  
V. G. Ivashenko
Keyword(s):  
Biological Control ◽  
Fusarium Head Blight ◽  
Fusarium Graminearum ◽  
Head Blight ◽  
Fusarium Spp

scholarly journals Population Structure of Fusarium graminearum Isolated from Different Sources in One Area over the Course of Three Years

2020 ◽  
Vol 110 (7) ◽  
pp. 1312-1318 ◽  
Author(s):  
Donatas Sneideris ◽  
Algirdas Ivanauskas ◽  
Petras Prakas ◽  
Dalius Butkauskas ◽  
Olga Treikale ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Population Analysis ◽  
Sensu Stricto ◽  
Variable Number ◽  
Plant Residue ◽  
Head Blight ◽  
Plant Host ◽  
Plant Matter ◽  
Control And Prevention ◽  
Different Sources

Fusarium head blight (FHB) is an important crop disease worldwide and is mainly caused by members of the Fusarium graminearum species complex. F. graminearum sensu stricto is the most common cosmopolitan and predominant FHB causal agent in Europe. Thus far, the majority of studies have focused on the primary hosts (wheat and barley) of this pathogen, while the relationships between other sources of infection remain unclear. We monitored and sampled test fields over the course of 3 years and acquired 804 F. graminearum isolates from different sources: primary hosts and other plant species included in the crop rotations, weeds from the test fields, decaying plant residue, soil samples, and crop seed. Of these isolates, 73.3% had the 15-acetyldeoxynivalenol genotype and 26.7% had the 3-acetyldeoxynivalenol genotype. F. graminearum isolation rates from weeds (>50%) were much higher than from soil (< 10%) or decaying plant matter (4%). Variable number of tandem repeat markers were used for population analysis. Noticeable genetic differentiation was detected between isolates from living plants and soil biome. In contrast, absence of any noticeable division between primary and alternative plant host communities indicates the importance of weeds and other segetal plants for FHB control and prevention.

scholarly journals Nivalenol-Type Populations of Fusarium graminearum and F. asiaticum Are Prevalent on Wheat in Southern Louisiana

2011 ◽  
Vol 101 (1) ◽  
pp. 124-134 ◽  
Author(s):  
Liane Rosewich Gale ◽  
Stephen A. Harrison ◽  
Todd J. Ward ◽  
Kerry O'Donnell ◽  
Eugene A. Milus ◽  
...  
Keyword(s):  
United States ◽  
North Carolina ◽  
Fusarium Graminearum ◽  
Gulf Coast ◽  
Pcr Primers ◽  
The United States ◽  
Sensu Stricto ◽  
Head Blight ◽  
Trichothecene Mycotoxin ◽  
Southern Louisiana

U.S. populations of the Fusarium graminearum clade cause head blight on wheat and barley and usually contaminate grain with the trichothecene mycotoxin deoxynivalenol (DON). Recently, however, individual nivalenol (NIV)-type isolates from the United States were described that belonged to either the newly described species F. gerlachii or the genetically distinct Gulf Coast population of F. graminearum sensu stricto (s.s.). Here, we describe the discovery of NIV-type F. graminearum s.s. populations that were found in high proportion (79%) among isolates from small-grain-growing regions of Louisiana. We genotyped 237 isolates from Louisiana with newly developed polymerase chain reaction (PCR) restriction fragment length polymorphism markers and multiplex PCR primers that distinguish among the three trichothecene types: the two DON types (15ADON and 3ADON) and NIV. These isolates were compared with 297 isolates from 11 other U.S. states, predominantly from the Midwest. Using Bayesian-model-based clustering, we discovered a southern Louisiana population of F. graminearum s.s. that was genetically distinct from the previously recognized pathogen population in the Midwest (MW15ADON population). Population membership was correlated with trichothecene type. Most isolates from the southern Louisiana population were of the NIV type, while the majority of the isolates from the Midwest were of the 15ADON type. A smaller proportion of isolates from Louisiana belonged to the previously described Gulf Coast population that was mostly of the 3ADON type. The NIV type was also identified in collections from Arkansas (12%), North Carolina (40%), and Missouri (2%), with the collections from Arkansas and North Carolina being small and unrepresentative. F. asiaticum was detected from the two southern Louisiana parishes Acadia and Alexandria. All identified 41 F. asiaticum isolates were of the NIV type. Greenhouse tests indicated that U.S. NIV types accumulated four times less trichothecene toxin than DON types on inoculated wheat. This is the first report of NIV-type populations of F. graminearum s. s. and F. asiaticum in the United States.

scholarly journals Within-Field Variation of Fusarium graminearum Isolates for Aggressiveness and Deoxynivalenol Production in Wheat Head Blight

2012 ◽  
Vol 102 (1) ◽  
pp. 128-134 ◽  
Author(s):  
Firas Talas ◽  
Rasha Kalih ◽  
Thomas Miedaner
Keyword(s):  
Fusarium Graminearum ◽  
Sensu Stricto ◽  
Wheat Breeding ◽  
Phenotypic Variance ◽  
Environment Interaction ◽  
Field Variation ◽  
Head Blight ◽  
Single Spore ◽  
Spring Wheat Cultivar ◽  
Multiple Resistance Genes

Fusarium head blight (FHB), caused by Fusarium graminearum sensu stricto (s.s.), causes tremendous annual yield losses in wheat worldwide. Variation of aggressiveness of isolates from individual field populations in terms of FHB infection and deoxynivalenol (DON) concentration in the host are important population parameters reflecting parasitic ability. Our main objective was to estimate the variation of both traits within three populations of F. graminearum s.s., each consisting of 30 single-spore isolates collected from small wheat fields in Germany, and to compare it with 11 isolates of a collection (F. graminearum collection) from four countries. The same isolates were characterized using 19 single-sequence repeat markers. All isolates were spray inoculated on a moderately resistant spring wheat cultivar at two field locations over 2 years (i.e., in four environments). The genotypic proportion of phenotypic variance (σ2G) within populations was significant (P < 0.01) for both traits, and the σ2G × environment interaction was even more important for mean FHB severity. Ranges in mean FHB severity and DON concentration in the host were only slightly smaller for the field populations than for the F. graminearum collection. Both traits were significantly (P < 0.05) correlated within and across populations. A further partitioning of σ2G revealed 72% of σ2G within and 28% of σ2G across populations for both traits. Molecular variance of the three populations was similarly distributed (73.6% within versus 26.4% between populations). In view of this high within-field variation for traits of parasitic ability and selection, neutral molecular markers, multiple resistance genes of different origin should be employed in wheat breeding programs to obtain a long-term stable FHB resistance.

scholarly journals First report of Head blight of wheat caused by Fusarium vorosii in Serbia

Plant Disease ◽  
2021 ◽  
Author(s):  
Ana Obradović ◽  
Jelena Stepanovic ◽  
Vesna Krnjaja ◽  
Aleksandra Bulajic ◽  
Goran Stanković ◽  
...  
Keyword(s):  
Fusarium Graminearum ◽  
Species Recognition ◽  
Histone H3 ◽  
Elongation Factor ◽  
Phylogenetic Species ◽  
Head Blight ◽  
Single Spore ◽  
First Report ◽  
Genealogical Concordance ◽  
Β Tubulin

The cosmopolitan species Fusarium graminearum Schwabe directly reduces yield, as well as grain quality of cereals, due to its ability to synthesize mycotoxins. Previously it was considered to be one species occurring on all continents. However, phylogenetic analysis employing the GCPSR method (Genealogical Concordance Phylogenetic Species Recognition) revealed the existence of 15 phylogenetic species within what is now recognised as the Fusarium graminearum Species Complex (FGSC) (Sarver et al. 2011). During 1996-2008, a MRIZP collection of FGSC isolates was established and isolates originating from wheat (5), maize (3) and barely (2) were selected for further study. Morphological features including the appearance of colonies and macroconidia (average size 38.5-53.1 × 4.6-5.4 µm, No 50) of all 10 isolates on PDA were consistent with descriptions of F. graminearum (O’Donnell et al. 2004, Leslie and Summerell 2006). Total DNA was isolated from mycelium removed from 7-day old colonies of single-spore isolates grown on PDA using the DNeasy Plant Mini Kit (Qiagen, Hilden). Further identification was based on amplification and sequencing of elongation factor TEF−1α, histone H3 and β−tubulin in both directions, with primers ef1/ef2, H3-1a/H3-1b and T1/T22, respectively (Jacobs et al. 2010). The sequences were deposited in NCBI under accession numbers MF974399 - MF974408 (TEF−1α), MG063783 - MG063792 (β−tubulin) and MF999139 - MF999148 (histone H3). Sequence analysis was performed using BLAST while genetic similarity was calculated using MEGA 6.0 software. Isolate 1339 originating from wheat (collected at the locality of Kikinda in 2006), shared 100% nucleotide identity with TEF−1α (DQ459745), histone H3 (DQ459728) and β−tubulin (DQ459643) of F. vorosii isolate NRRL37605 (Starkey et al. 2007). The remaining nine isolates were identified as F. graminearum as they shared 99% to 100% nucleotide similarity with F. graminearum NRRL 28439 (O’Donnell et al. 2004). Pathogenicity was tested using artificial inoculations of spikes during wheat flowering (Mesterhazy et al. 1999). Thirty classes were inoculated with each isolate, in three replicates. Inoculum was prepared from 7-day colonies on PDA, and 30 ml of a conidia suspension (1x105 conidia/ml) was used. Control plants were inoculated with sterile water. Three weeks after inoculation, typical Fusarium head blight symptoms were visible on inoculated plants, from which all 10 isolates were successfully reisolated. Control spikes remained symptomless. Disease severity was estimated on the 1-7 scale (Blandino et al. 2012). Average pathogenicity of the F. vorosii isolate 1339 was 1.9, and 2.4 -5.1 of F. graminearum isolates. Toxin production was determined using gas chromatography-tandem mass spectrometry. Kernels inoculated with the 10 isolates were ground and tested for the presence of deoxynivalenol (DON) and its acetyl derivatives 3ADON, 15ADON and NIV. F. vorosii isolate 1339 possessed the 15ADON chemotype, as well as eight F. graminearum isolates, while only one F. graminearum isolate was 3ADON chemotype. To date, F. vorosii has only been detected in Hungary on wheat (Toth et al. 2005) and Korea on barley, corn and rice (Lee et al. 2016). This is the first report of F. vorosii in Serbia, which is of great importance, because it indicates the spread of this toxigenic species. Further studies should be focused on determining the distribution, aggressiveness and toxicological profile of F. vorosii.

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