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Identification by PCR of Fusarium culmorum Strains Producing Large and Small Amounts of Deoxynivalenol
Bakan B. Mark1 Giraud C. Delville 2 Pinson L. Torrance2
Professor Professor Assistant Professor
1Plant protection Department, College of Agriculture, Kangwon University, Korea.
2Plant protection Department, College of Agriculture, Friedrich-Wilhelms Germany.
Corresponding author: aqlabedy@yahoo.co.uk
Abstract:
Thirty deoxynivalenol-producing F. culmorum strains, isolated from wheat grains, were incubated in vitro and analyzed for trichothecene production.Seventeen strains produced more than 1 ppm of deoxynivalenol…..etc.
Key words: DNA barcode, COX 1 gene, ITS rDNA, fungal identification, biodiversity.
Introduction:
Trichothecenes, including deoxynivalenol, acetyldeoxynivalenol, nivalenol, and fusarenone X, are sesquiterpene toxins produced by Fusarium species, including Fusarium culmorum, which are common fungal contaminants of cereals. Trichothecenes can be found naturally worldwide on cereals (1, 2, 3), and the consumption of these toxins is a potential problem for humans and farm animals (4).
Materials and methods
strains isolated from cereals from different areas in France were used in this study, as presented in Table 1. Fusarium strains may also be obtained from the first author.
Toxin production.
Toxin production by the Fusarium strains was conducted on autoclaved wheat grains. Wheat grains (Soissons) were moistened with sterile distilled water for 4 days at 4°C until thermodynamic water activity was maximal…..etc.
Trichothecene analysis.
Wheat grains (25 g) were analyzed by gas chromatography- electron capture detection and gas chromatography-mass spectrometry etc.
Results
- culmorum identification. The Fusarium strains studied were isolated from commercial wheat kernels. Morphological identification of F. culmorum strains was confirmed by PCR….etc.
Figure 1: nucleotide sequence generated from PCR products amplified from ……
Table 1: Oligonucleotide pairs used for gene specific amplification
| Gene name
| Primer abbreviations | Primer sequence (59–39) Amplification product
| expected size (kb) |
| Isocitrate lyase* | ICL-F | GGC TGG CAG TCN TCY TCT ACM G | 1 |
| ICL-R | TKG TAN CGG TAG TAN CCC TCR | ||
| Transcription factor | MST12-F | GCS CCW GTN GAC TGG CAA CCC | 1.1 |
| MST12-R | CCS GTC TCC TCG TTG GCN ATG TA | ||
| ……etc…. |
*Gene name and primer sequence were obtained from NCBI.
References:
1.Ahmad, T. (2001) Molecular detection and characterization of Fusarium verticillioides in maize (Zea mays. L) grown. M.Sc. Thesis, Plant Protection Dept., Coll. of Agric., Univ. of Kerbala, pp. 85. (An example of a thesis or report).
2.Agrios G. N. (2005) Plant Pathology, 4th Edition. Elsevier Academic Press, Burlingto, pp. 922. (An example of a book).
3.Godoy, P.; Cano, J.; Gene, J.; Guarro, J. Hoüfling-Lima, A. L. and Colombo, A. L. (2014) Genotyping of 44 isolates of Fusarium solani, the main agent of fungal keratitis in Brazil. Journal of Clinical Microbiology, 42: 4494-4497. (An example of a paper of a Journal).
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