Genetic characterization of Fusarium graminearum and F. culmorum isolates from Turkey by using random-amplified polymorphic DNA

Yörük, Emre; Albayrak, Gülruh

doi:10.4238/2013.april.25.7

Cited by 14 publications

(6 citation statements)

References 26 publications

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“…The CRED-RA method was used in the epigenetic profiling of camphor-treated and non-treated F. graminearum and F. culmorum . A routine RAPD protocol [39] was used in GTS assays of MspI -digested, HapII -digested, and non-digested control experimental sets. In total, 15 RAPD primers were used in this study (Table 2).…”

Section: Methodsmentioning

confidence: 99%

Investigation of Camphor Effects on Fusarium graminearum and F. culmorum at Different Molecular Levels

et al. 2018

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Fusarium graminearum and F. culmorum are phytopathogens, which cause destructive diseases in cereals. Epidemics of these phytopathogens are caused by mycotoxin contamination and the reduction of crop quality. In this study, the alteration due to in vitro camphor treatment on F. culmorum 9F and F. graminearum H11 isolates was investigated in terms of epigenetic, cellular, and transcription levels. Camphor with different concentrations (0.2, 0.4, 0.8, 1, 2, and 4 µg/µL) was applied to potato dextrose agar (PDA) growth media. The minimum inhibitory concentration (MIC) and the half maximal inhibitory concentration (IC50) were calculated as 2 and 1 µg/µL, respectively. hog1, mst20, CAT, POD, mgv1, stuA, and tri5 genes, which are related to various cellular processes and pathogenesis, were examined by qPCR assay. qPCR analysis showed that camphor treatment leads to the downregulation of tri5 expression but the upregulation of the remaining genes. Apoptosis and oxidative stress were confirmed via acridine orange/ethidium bromide (AO/EB) and dichlorofluorescin diacetate (DCF-DA) staining, respectively. Moreover, coupled restriction enzyme digestion-random amplification (CRED-RA) assay, used for DNA methylation analysis, was carried out to evaluate epigenetic alterations. The decrease in genomic template stability (GTS) values, which resulted due to the alterations in random amplified polymorphic DNA (RAPD) profiles caused by camphor treatment, were detected as 97.60% in F. culmorum 9F and 66.27% in F. graminearum H-11. The outer and inner methylated cytosine profiles are determined by CRED-RA assay as type I–IV epigenetic alterations. The outcomes indicated that camphor could lead to alterations at several molecular levels of F. graminearum and F. culmorum.

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Section: Methodsmentioning

confidence: 99%

Investigation of Camphor Effects on Fusarium graminearum and F. culmorum at Different Molecular Levels

et al. 2018

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“…RAPD and CRED-RA methods were used in the genomic template stability and epigenetic profiling of TCZ treated and non-treated F. graminearum. A routine RAPD protocol (Yörük and Albayrak, 2013) was used in GTS assays of non-digested control, MspI-digested, HapII-digested experiment sets by using thermal cycler (Hi-Media, India). The total of 20 RAPD primers were used in this study (Table 1).…”

Section: Genomic Dna (Gdna) Extraction Rapd (Random Amplified Polymorphic Dna) and Cred-ra (Coupled Restriction Enzyme Digestion-random Amentioning

confidence: 99%

Tetraconazole Leads to Alterations in Fusarium Graminearum at Different Molecular Levels

Yörük¹

2018

Appl. Ecol. Env. Res.

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The alterations in F. graminearum due to tetraconazole (TCZ) have been investigated in this study. The minimum inhibitory concentration (MIC), and inhibitory concentrations 50% and 25% (IC 50 and IC 25 ) were obtained by adding different concentrations of TCZ to potato dextrose agar (PDA). MIC, IC 50 and IC 25 values were detected as 32, 16 and 8 µg/mL TCZ. Epigenetic changes have been evaluated via genomic template stability (GTS) by the RAPD and CRED-RA methods. GTS values were recorded as 86.73% and 85.71% in IC 50 and IC 25 sets. A total of 157 bands were obtained via RAPD. The average % polymorphism values of HapII and MspI digested samples were detected as 5.72 and 5.37 with 4.68 and 2.6% for IC 25 and IC 50 groups, respectively. The expression levels of genes related to apoptosis (Hog1), cell stability (Mgv1), oxidative stress (POD) and deoxynivalenol production (tri5) in the control and the experiment sets were investigated by qPCR. Increased concentrations of TCZ lead to upregulation in Hog1, Mgv1 and POD genes whereas down regulation was recorded in tri5 expression. The late apoptosis and oxidative stress were detected via the acridine orange-ethidium bromide (AoEb) and the DCF-DA staining assays. The findings showed that TCZ could lead to damage on phytopathogenic fungi at genomic, epigenetics, transcriptomics and apoptotic levels.

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“…Similarly, isolates from the same or near agroecological location (s) were clustered so closely in phylogram. Even if the precise correlation between geographic origin and clustering of Fusarium isolates can't be stated whenever, locations can be accepted as a major characteristic behind the variation of Fusarium isolates (Carter et al, 2000(Carter et al, , 2002Mishra et al, 2004;Miedaner et al, 2001Miedaner et al, , 2008Yörük and Albayrak, 2013). Most of the isolates used in this study were obtained from wheat as the host.…”

Section: Resultsmentioning

confidence: 99%

First Report on Identification of Fusarium graminearum Species Complex Members from Turkey and Iran

Tunalı

Yörük

Sefer

et al. 2019

Turkish JAF Sci.Tech.

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Fusarium graminearum species complex is the major Fusarium head blight disease pathogen in worldwide. Fusarium head blight disease lead to damages on small grain cereals. The identification of the F. graminearum species complex is important for improving disease control and management. Traditional identification methods such as macroscopy and microscopy analysis need to be supported by biochemical and genetics assays. Thus, DNA sequencing-based methods is one of the most preferred, reliable, low priced in the identification methods. In the study, 54 F. graminearum isolates obtained from diseased wheat, barley and corn fields in Turkey and Iran were identified by morphological characteristics and then characterized by species-specific SCAR marker. β-tubulin, Tef1-α, 28s rDNA and Histone H3 genes amplified, purified and then sequenced. The merged multiloci length was obtained as last of all 2215 bp. These sequencing results was used to multiloci genotyping assays. Last of all, 20 isolates were determined as F. graminearum sensu stricto by multiloci genotyping analysis. Remaining isolates were identified as F. asiaticum or Fusarium sp. . The findings are important in terms of revealing the first-time identification in Turkish and Iranian isolates as F. graminearum species complex members by amplification four (β-tubulin, Tef1-α, 28s rDNA and Histone H3) highly conserved DNA regions.

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Genetic characterization of Fusarium graminearum and F. culmorum isolates from Turkey by using random-amplified polymorphic DNA

Cited by 14 publications

References 26 publications

Investigation of Camphor Effects on Fusarium graminearum and F. culmorum at Different Molecular Levels

Investigation of Camphor Effects on Fusarium graminearum and F. culmorum at Different Molecular Levels

Tetraconazole Leads to Alterations in Fusarium Graminearum at Different Molecular Levels

First Report on Identification of Fusarium graminearum Species Complex Members from Turkey and Iran

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