Fusarium species and Deoxynivalenol in maize product of Moqan region

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1 DOI: Fusarium species and Deoxynivalenol in maize product of Moqan region - - ( : : ) -... (DON). Czapek PDA Nash-Snyder CLA PDA. % / F. proliferatum % / F. verticillioides : DON.% / F. oxysporum % /. (IAC) (HPLC) F. nygamai Corresponding author:

2 ... :. % ng/g DON / / (1 ppm) ng/g / DON. ( ) - F. verticillioides F. oxysporum F. nygamai. DON. DON F. proliferatum DON F. proliferatum. DON. DON. : Abstract Ear rot of corn caused by Fusarium species could create an overall comprehensive problem. Fusarium species can produce mycotoxins such as trichothecenes which threat the human's health. Cultivation of maize in Moqhan region was about hectares. Forty (5-10 Kg) samples each including 10 subsamples of corn ears collected at harvest time, kernels separated from the ears, dried and divided into two parts: one part for mycological studies and another part for toxicology studies. The media Nash-Snyder, PDA and Czapek were used for isolation, and PDA and CLA for identification. Deoxynivalenol detection and measurement of the corn samples were carried out using IAC - HPLC (High Performance Liquid Chromatography and immunoaffinity Columns). To study the potential of deoxynivalenol production of the isolates, ten isolates (two of each species) were selected as the representative of each species, the potential of DON production for each isolate was measured after one week incubation at 25-27ºC and two weeks at 12ºC alternatively. To ensure the deoxynivalenol production of the two F. proliferatum isolates, these isolates inoculated artificially in corn ears and after 10 days the amount of DON was detected in the grains. Fusarium species and the frequency of isolation were F. verticillioides, 47.65%;

3 : F. proliferatum, %, 15.33%; F. nygamai, 4.86%; F. oxysporum,3.06. Deoxynivalenol was detected in 45% of the samples. The total mean of contamination was 45% and the range of contamination was ng/g with the mean of ng/g. Among the Fusarium species isolates two isolates of F. proliferatum could produce DON in artificial conditions. Key words: deoxynivalenol, Fusarium species, corn..." ". (Gibberella ear rot) (Munkvold, 2003) (Fusarium ear rot).(menna et al., 1997) Fusarium graminearum. ( ).(Sutton, 1982) () F. culmorum.(tschanz et al., 1976).(Munkvold, 2003) F. verticillioides...

4 ... :. (DON).(Schollenberger et al., 2002; Schaafsma et al., 2001).(Abbas et al., 1988). DON.(Schaafsma et al., 2001)... DON DON. DON : -...( ).. DON

5 : - Fig 1- Infected corn samples Potato Dextrose Agar Nash-Snyder PPA 7 H 2 O :(Pepton PCNB Agar) Nash-Snyder KH 2 PO 4 : - Pepton.(Nelson et al., 1983) - - : (PCNB / ). Agar Teraclor MgSO 4 /. :(PDA) CLA SNA : - PDA.(Nelson et al., 1983) PDA. -.

6 ... :. PDA. (Nelson et al., 1983). (Nelson et al., 1983; Leslie and summerell, 2006) : - : - -. (Romer) - -. ( ).. (Whatman International Ltd, England) (GF/A glass microfibre ). (R-Biopharm) (Clean up) DONprep ) (. / (Rotary evaporator).(mirabolfathy and Karami Osboo, 2006) ( : : ) HPLC HPLC. : HPLC - - (binary HPLC pump 1525) Breeze (HPLC)

7 : UV Waters. 4 µm mm C-18 (Waters Milford, MA, USA) ( 6:6:88) : : (UV Detector) UV.. Breeze 222 nm. :(Calibration curve) - - HPLC.. (Regression).( ) (R 2 =0.999) ( - ) - Fig. 2. Deoxynivalenol calibration curve ( ng/ml)

8 ... : 250 ng/ml - Fig 3- The Chromatogram of Deoxynivalenol standard (250 ng/ml).. : - - µg/ml 2.5ml. (Recovery). ppm (spike).( ). HPLC %.

9 : - Fig 4- The chromatogram of the blank sample ppb - Fig 5- The chromatogram of 1000 ng/ml spiked sample : - - ) (.

10 ... :. / ) - ( F. proliferatum DON. : - :.F. oxysporum F. nygamai F. proliferatum Fusarium verticillioides : -.

11 : - Table 1- The frequency of Fusarium species isolated from each corn samples F. oxysporum F. nygamai F. verticioiledes F. proliferatum samples M M M M M M M M M M M M M M M M M M M M M M M M M M M9-8 4 M M M M M M M M M M M M M Total

12 ... : - Table 2 -The frequency (%)of Fusarium species isolated from corn kernels Fusarium species Frequency Fusarium verticillioides F. proliferatum F. nygamai F. oxysporum (Placinta et al., 1999).( ) % / ng/g / -/. (1 ppm) ng/g : - - ppm..

13 : ( ). : - -,F. nygamai F. verticillioides F. oxysporum.( ) F. proliferatum - Table 3- DON contamination (ppb) of corn samples DON contamination (ppb) Samples DON contamination (ppb) Samples M3 M4 M5 M6 M7 M8 M9 M10 M11 M12 M13 M14 M15 M16 M17 M18 M19 M20 M21 M M001 M002 M003 M004 M005 M006 M007 M008 M009 M010 M011 M012 M013 M014 M015 M016 M017 M018 M1 M2 : Not Determinated

14 ... :. DON F. proliferatum DON.( ) / / ppb - Table 4- Distribution of DON concentration in contaminated samples Contaminated samples (%) Don concentration (ppb) Table 5 - The potential of DON production in different Fusarium species isolates on rice powder media Sample number Isolate F. oxysporum F. oxysporum F. nygamai F. nygamai F. verticillioides F. verticillioides F. proliferatum F. proliferatum (ppb) Potential of deoxynevalenol

15 : F. proliferatum (ppb) DON - Table 6 -DON concentration detected (ppb) in the inoculated corn ears by F. proliferatum isolates Inoculated corn ears DON concentration (ppb) 1 2 F. proliferatum F. proliferatum F. proliferatum. F. verticillioides % / Liseola F. nygamai. F. oxysporum. Elegans F. graminearum.. F. verticillioides F. verticillioides F. graminearum.(stewart et al., 2002) F. verticillioides (Boujari and Ershad, 1993) F. verticillioides F. graminearum F. verticillioides F. graminearum F. verticillioides. F. graminearum.

16 ... :. DON.. F. proliferatum F. proliferatum F. verticillioides..f. graminearum F. nygamai F. oxysporum F. proliferatum F. verticillioides. F. graminearum F. proliferatum F. verticillioides. F. graminearum ) F. graminearum (.. F. verticillioides. F. graminearum F. graminearum F. graminearum. F. verticillioides. F. verticillioides.(stewart et al., 2002) F. graminearum

17 :.. ABBAS, H. K., C. J. MIROCHA, R. A. MERONUCK, J. D. POKORNY, S. L. GOULD and T. KOMMEDAHL, Mycotoxins and Fusarium spp. associated with infected ears of corn in Minnesota. Appl. Environ. Microbiol. 54: Agronomic considerations for reducing deoxynivalenol in wheat grain. Canadian Journal of Plant Pathology. 23: BOUJARI, J. and D. ERSHAD, An Investigation on Corn - seed Mycoflora. Iranian Journal of Plant Pathology, 29: LESLIE, F. and B. SUMMERELL, The Fusarium Laboratory Manual MENNA DI, M. E., D. R. LAUREN and A. HARDACRE, Fusaria and Fusarium toxins in New Zealand maize plants. Mycopathologia. 139: MIRABOLFATHY, M. and R. KARAMI OSBOO, Monitoring of deoxynivalenol (DON) in wheat crop at Golestan province. Proceedings of the 17 th Iranian Plant Protection Congress. P. 505 MUNKVOLD, G. P Epidemiology of Fusarium diseases and their mycotoxins in maize ears. European Journal of Plant Pathology. 109: NELSON, E. T., A. Toussoun and W. F. O. Marasas, Fusarium Species: an illustrated manual for identification. Pennsylvania State University Press, 193 pages PLACINTA, C. M., J. P. F. D MELLO and A. M. C. MAC DONALD, A review of :.

18 ... : worldwide contamination of cereal grains and animal feed with Fusarium mycotoxins. Anim. Feed Sci. Techno. 78: SCHAAFSMA, A. W., L. TAMBURIC-ILLNIC, J. D. MILLER, D. C. HOOKER, SCHOLLENBERGER, M., H. T. JARA, S. SUEY, W. DROCHNER, H. M. MGLLER, Fusarium toxins in wheat flour collected in an area in southwest Germany. International Journal of Food Microbiology, 72: SCIENTIFIC COMMITTEE ON FOOD, Opinion on Fusarium Toxins-Part 1: Deoxynivalenol (DON) (expressed on 2 December 1999). Available at intlcomm/food/fs/sc/scf/out44_en.pdf STEWART, D. W., L. M. REID, R. W. NICOL and A. W. SCHAAFSMA, A mathematical simulation of growth of Fusarium in maize ears after artificial inoculation. Phytopathology, 92: SUTTON, J. C Epidemiology of wheat head blight and maize ear rot caused by Fusarium graminearum. Canadian Journal of Plant Pathology. 4: TSCHANZ, A. T., R. K. HORST and P. E. NELSON, The effect of environment on sexual reproduction of Gibberella zeae. Mycologia 68: Address of the authors: Eng. Z. ALIAKBARI and Dr. H. AMINIAN, Department of Plant Protection, Faculty of Agriculture, Tehran University, Karaj, Iran; Dr. M. MIRABOLFATHY and Eng. R. KARAMI OSBOO, Iranian Research Institute of Plant Protection, P. O. Box 1454, Tehran, Iran.

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