Mapping of genes expressed in Fusarium graminearum-infected heads of wheat cultivar 'Frontana'

Genome ◽  
2005 ◽  
Vol 48 (1) ◽  
pp. 88-96 ◽  
Author(s):  
F P Han ◽  
G Fedak ◽  
T Ouellet ◽  
H Dan ◽  
D J Somers

The isolation, physical, and genetic mapping of a group of wheat genes expressed in infected heads of Triticum aestivum 'Frontana' resistant to Fusarium head blight is reported. A cDNA library was built from heads of 'Frontana' through suppressive subtractive hybridization, to enrich for sequences induced by the pathogen Fusarium graminearum during infection. A group of 1794 clones was screened by dot blot hybridization for differential gene expression following infection. Twenty of these clones showed a strong difference in intensity of hybridization between infected and mock-inoculated wheat head samples, suggesting that they corresponded to genes induced during infection. The 20 clones were sequenced and used for mapping analysis. We determined a precise chromosomal location for 14 selected clones by using series of chromosome deletion stocks. It was shown that the 14 clones detected 90 fragments with the use of the restriction enzyme EcoRI; 52 bands were assigned to chromosome bins, whereas 38 fragments could not be assigned. The selected clones were also screened for polymorphisms on a 'Wuhan' × 'Maringa' wheat doubled haploid mapping population. One clone, Ta01_02b03, was related to a quantitative trait locus for type II resistance located on chromosome 2AL, as determined with simple sequence repeat markers on another mapping population, but did not map in the same location on our population. Another clone, Ta01_06f04, was identified by BLAST (basic local alignment search tool) search in public databases to code for a novel β-1,3-glucanase, homologous to a major pathogenesis-related protein. This clone mapped to chromosomal regions on chromosome 3, including 3BL and 3DL, where B glucanase gene clusters are known to exist. Seven other clones, including 1 coding for an ethylene-response element binding protein and 3 for ribosomal proteins, and 4 clones corresponding to proteins with unknown function, were also mapped.Key words: deletion stock, Fusarium head blight, physical mapping, expressed sequence tags, glucanase.

Genome ◽  
2005 ◽  
Vol 48 (1) ◽  
pp. 29-40 ◽  
Author(s):  
Lingrang Kong ◽  
Joseph M Anderson ◽  
Herbert W Ohm

Fusarium head blight (FHB), caused by species of the fungus Fusarium, is a worldwide disease of wheat (Triticum aestivum L.). The Chinese T. aestivum 'Ning7840' is one of few wheat cultivars with resistance to FHB. To identify differentially expressed genes corresponding to FHB resistance, a cDNA library was constructed using pooled mRNA isolated from glumes of 'Ning7840' harvested at 2, 6, 12, 24, 36, 72, and 96 h after inoculation (hai) with a conidia spore suspension of Fusarium graminearum. Suppressive subtractive hybridization (SSH) cDNA subtraction was carried out using pooled glume mRNAs from the tester and the control. The cDNA library was differentially screened using the forward subtracted cDNAs and the reverse subtracted cDNAs as probes. Twenty-four clones with significant matches to either plant (16 sequences) or fungal (8 sequences) genes were isolated based on their specific hybridization with forward subtracted cDNA and not reverse subtracted cDNA. Six putative defense-related genes were confirmed by real-time quantitative reverse-transcriptase PCR. Many-fold higher induction of three clones (A3F8, B10H1, and B11H3) in the resistant genotypes compared with susceptible genotypes indicates a putative role in the resistance response to Fusarium graminearum. Transcript accumulations of P450, chitinase (Chi1), and one unknown gene (clone B8Q9) in both resistant and susceptible genotypes suggest an involvement in a generalized resistance response to F. graminearum. Nucleotide sequence analysis showed that cDNA clone A4C6 encodes a cytochrome P450 gene (CYP709C3v2), including 14 N-terminal amino acids that have a membrane-associated helical motif. Other domains characteristic of eukaryotic P450 are also present in CYP709C3v2. The deduced polypeptide of cDNA clone B2H2 encodes an acidic isoform of class I chitinase containing a 960-bp coding region. Southern hybridization using aneuploid lines of T. aestivum 'Chinese Spring' indicated that CYP709C3v2 was located on the short arm of chromosomes 2B and 2D.Key words: Fusarium head blight (FHB), suppressive subtractive hybridization, defense response, real-time quantitative RT-PCR.


BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Thomas E. Witte ◽  
Linda J. Harris ◽  
Hai D. T. Nguyen ◽  
Anne Hermans ◽  
Anne Johnston ◽  
...  

Abstract Background Fusarium head blight is a disease of global concern that reduces crop yields and renders grains unfit for consumption due to mycotoxin contamination. Fusarium poae is frequently associated with cereal crops showing symptoms of Fusarium head blight. While previous studies have shown F. poae isolates produce a range of known mycotoxins, including type A and B trichothecenes, fusarins and beauvericin, genomic analysis suggests that this species may have lineage-specific accessory chromosomes with secondary metabolite biosynthetic gene clusters awaiting description. Methods We examined the biosynthetic potential of 38 F. poae isolates from Eastern Canada using a combination of long-read and short-read genome sequencing and untargeted, high resolution mass spectrometry metabolome analysis of extracts from isolates cultured in multiple media conditions. Results A high-quality assembly of isolate DAOMC 252244 (Fp157) contained four core chromosomes as well as seven additional contigs with traits associated with accessory chromosomes. One of the predicted accessory contigs harbours a functional biosynthetic gene cluster containing homologs of all genes associated with the production of apicidins. Metabolomic and genomic analyses confirm apicidins are produced in 4 of the 38 isolates investigated and genomic PCR screening detected the apicidin synthetase gene APS1 in approximately 7% of Eastern Canadian isolates surveyed. Conclusions Apicidin biosynthesis is linked to isolate-specific putative accessory chromosomes in F. poae. The data produced here are an important resource for furthering our understanding of accessory chromosome evolution and the biosynthetic potential of F. poae.


Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 68
Author(s):  
Gaetano Bentivenga ◽  
Alfio Spina ◽  
Karim Ammar ◽  
Maria Allegra ◽  
Santa Olga Cacciola

In 2009, a set of 35 cultivars of durum wheat (Triticum turgidum L. subsp. durum (Desf.) Husn.) of Italian origin was screened for fusarium head blight (FHB) susceptibility at CIMMYT (Mexico) and in the 2019–20 cropping season, 16 of these cultivars, which had been included in the Italian National Plant Variety Register, were tested again in southern and northern Italy. Wheat cultivars were artificially inoculated during anthesis with a conidial suspension of Fusarium graminearum sensu lato using a standard spray inoculation method. Inoculum was a mixture of mono-conidial isolates sourced in the same areas where the trials were performed. Isolates had been characterized on the basis of morphological characteristics and by DNA PCR amplification using a specific primer set and then selected for their virulence and ability to produce mycotoxins. The susceptibility to FHB was rated on the basis of the disease severity, disease incidence and FHB index. Almost all of the tested cultivars were susceptible or very susceptible to FHB with the only exception of “Duprì”, “Tiziana” and “Dylan” which proved to be moderately susceptible. The susceptibility to FHB was inversely correlated with the plant height and flowering biology, the tall and the late heading cultivars being less susceptible.


2008 ◽  
Vol 88 (6) ◽  
pp. 1087-1089 ◽  
Author(s):  
Stephen N Wegulo ◽  
Floyd E Dowell

Fusarium head blight (scab) of wheat, caused by Fusarium graminearum, often results in shriveled and/or discolored kernels, which are referred to as Fusarium-damaged kernels (FDK). FDK is a major grain grading factor and therefore is routinely determined for purposes of quality assurance. Measurement of FDK is usually done visually. Visual sorting can be laborious and is subject to inconsistencies resulting from variability in intra-rater repeatability and/or inter-rater reliability. The ability of a single-kernel near-infrared (SKNIR) system to detect FDK was evaluated by comparing FDK sorted by the system to FDK sorted visually. Visual sorting was strongly correlated with sorting by the SKNIR system (0.89 ≤ r ≤ 0.91); however, the SKNIR system had a wider range of FDK detection and was more consistent. Compared with the SKNIR system, visual raters overestimated FDK in samples with a low percentage of Fusarium-damaged grain and underestimated FDK in samples with a high percentage of Fusarium-damaged grain. Key words: Wheat, Fusarium head blight, Fusarium-damaged kernels, single-kernel near-infrared


2012 ◽  
Vol 33 (1) ◽  
pp. 97-111 ◽  
Author(s):  
Johann Leplat ◽  
Hanna Friberg ◽  
Muhammad Abid ◽  
Christian Steinberg

2020 ◽  
Vol 13 (2) ◽  
pp. 235-246
Author(s):  
W.Q. Shi ◽  
L.B. Xiang ◽  
D.Z. Yu ◽  
S.J. Gong ◽  
L.J. Yang

Fusarium graminearum causes Fusarium head blight (FHB), a devastating disease that leads to extensive yield and quality loss in wheat and barley production. Integrated pest management (IPM) is required to control this disease and biofungicides, such as tetramycin, could be a novel addition to IPM strategies. The current study investigated in vitro tetramycin toxicity in Fusarium graminearum and evaluated its effectiveness for the control of Fusarium head blight FHB. Tetramycin was shown to affect three key aspects of Fusarium pathogenicity: spore germination, mycelium growth and deoxynivalenol (DON) production. The in vitro results indicated that tetramycin had strong inhibitory activity on the mycelial growth and spore germination. Field trials indicated that tetramycin treatment resulted in a significant reduction in both the FHB disease index and the level of DON accumulation. The reduced DON content in harvested grain was correlated with the amount of Tri5 mRNA determined by qRT-PCR. Synergistic effects between tetramycin and metconazole, in both the in vitro and field experiments were found. Tetramycin could provide an alternative option to control FHB.


Toxins ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 460 ◽  
Author(s):  
Vesna Krnjaja ◽  
Slavica Stanković ◽  
Ana Obradović ◽  
Tanja Petrović ◽  
Violeta Mandić ◽  
...  

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.


2014 ◽  
Vol 104 (11) ◽  
pp. 1201-1207 ◽  
Author(s):  
Luca Sella ◽  
Katia Gazzetti ◽  
Carla Castiglioni ◽  
Wilhelm Schäfer ◽  
Francesco Favaron

Fusarium graminearum is a toxigenic fungal pathogen that causes Fusarium head blight (FHB) and crown rot on cereal crops worldwide. This fungus also causes damping-off and crown and root rots at the early stage of crop development in soybean cultivated in North and South America. Several F. graminearum genes were investigated for their contribution to FHB in cereals but no inherent study is reported for the dicotyledonous soybean host. In this study we determined the disease severity on soybean seedlings of five single gene disrupted mutants of F. graminearum, previously characterized in wheat spike infection. Three of these mutants are impaired on a specific function as the production of deoxynivalenol (DON, Δtri5), lipase (ΔFgl1), and xylanase (Δxyl03624), while the remaining two are MAP kinase mutants (ΔFgOS-2, Δgpmk1), which are altered in signaling pathways. The mutants that were reduced in virulence (Δtri5, ΔFgl1, and ΔFgOS-2) or are avirulent (Δgpmk1) on wheat were correspondently less virulent or avirulent in soybean seedlings, as shown by the extension of lesions and seedling lengths. The Δxyl03624 mutant was as virulent as the wild type mirroring the behavior observed in wheat. However, a different ranking of symptom severity occurred in the two hosts: the ΔFgOS-2 mutant, that infects wheat spikelets similarly to Δtri5 and ΔFgl1 mutants, provided much reduced symptoms in soybean. Differently from the other mutants, we observed that the ΔFgOS-2 mutant was several fold more sensitive to the glyceollin phytoalexin suggesting that its reduced virulence may be due to its hypersensitivity to this phytoalexin. In conclusion, lipase and DON seem important for full disease symptom development in soybean seedlings, OS-2 and Gpmk1 MAP kinases are essential for virulence, and OS-2 is involved in conferring resistance to the soybean phytoalexin.


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