DOI: http://dx.doi.org/10.22092/jaep.2012.107219 Fusarium species and Deoxynivalenol in maize product of Moqan region - - ( : : ) n.aliakbari@gmail.com. -... (DON). Czapek PDA Nash-Snyder CLA PDA. % / F. proliferatum % / F. verticillioides : DON.% / F. oxysporum % /. (IAC) (HPLC) F. nygamai Corresponding author: n.aliakbari@gmail.com
... :. % 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 10000 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%;
: F. proliferatum, 33.873%, 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 59.4 542.55ng/g with the mean of 95.30. 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...
... :. (DON).(Schollenberger et al., 2002; Schaafsma et al., 2001).(Abbas et al., 1988). DON.(Schaafsma et al., 2001)... DON DON. DON : -...( ).. DON
: - 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. -.
... :. 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)
: UV Waters. 4 µm 250 3.9 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 (25-1000 ng/ml)
... : 250 ng/ml - Fig 3- The Chromatogram of Deoxynivalenol standard (250 ng/ml).. : - - µg/ml 2.5ml. (Recovery). ppm (spike).( ). HPLC %.
: - Fig 4- The chromatogram of the blank sample ppb - Fig 5- The chromatogram of 1000 ng/ml spiked sample : - - ) (.
... :. / ) - (... -... F. proliferatum DON. : - :.F. oxysporum F. nygamai F. proliferatum Fusarium verticillioides : -.
: - Table 1- The frequency of Fusarium species isolated from each corn samples F. oxysporum F. nygamai F. verticioiledes F. proliferatum samples - - 7 8 M001 - - 12 6 M002-2 4 6 M003-12 - M004 3-7 5 M005 - - 9 6 M006 - - 12 - M007 - - 6 4 M008-13 5 M009 - - 5 5 M010 - - 10 5 M011 - - 9 3 M012-3 9 M013 5-8 5 M014 - - 8 4 M015-6 4 - M016 - - 10 5 M017 - - 10 - M018 5 4-6 M1 - - 10 2 M2 - - - 10 M3 - - 9 5 M4 - - 8 2 M5 - - 9 6 M6 1-14 3 M7 - - 5 5 M8-5 7 - M9-8 4 M10 - - 5 5 M11 - - 6 9 M12 - - 8 6 M13 - - 12 - M14 - - 14 6 M15 - - 13 5 M16-5 5 5 M17-10 7 M18 - - 5 10 M19 6 - - 6 M20-5 10 5 M21 - - 13 5 M22 20 27 320 188 Total
... : - Table 2 -The frequency (%)of Fusarium species isolated from corn kernels Fusarium species Frequency Fusarium verticillioides F. proliferatum F. nygamai F. oxysporum 57.65 33.87 4.86 3.60....(Placinta et al., 1999).( ) % / ng/g / -/. (1 ppm) ng/g : - - ppm..
: ( ). : - -,F. nygamai F. verticillioides F. oxysporum.( ) F. proliferatum - Table 3- DON contamination (ppb) of corn samples DON contamination (ppb) Samples DON contamination (ppb) Samples 59.4 62.3 237.2 213 83.3 M3 M4 M5 M6 M7 M8 M9 M10 M11 M12 M13 M14 M15 M16 M17 M18 M19 M20 M21 M22 2288.92 451.43 66.290 48.141 280.08 542.55 131.5 261.4 192 151.2 224.7 156.6 M001 M002 M003 M004 M005 M006 M007 M008 M009 M010 M011 M012 M013 M014 M015 M016 M017 M018 M1 M2 : Not Determinated
... :. DON F. proliferatum DON.( ) / / ppb - Table 4- Distribution of DON concentration in contaminated samples Contaminated samples (%) 7.5 12.5 17.5 2.5 2.5 2.5 Don concentration (ppb) 0-100 100-200 200-300 300-400 400-500 500-600 - Table 5 - The potential of DON production in different Fusarium species isolates on rice powder media Sample number 1 2 3 4 5 6 7 8 Isolate F. oxysporum F. oxysporum F. nygamai F. nygamai F. verticillioides F. verticillioides F. proliferatum F. proliferatum (ppb) Potential of deoxynevalenol 62.6 57.4
: 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 30.2 32.4 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.
... :. 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
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... : worldwide contamination of cereal grains and animal feed with Fusarium mycotoxins. Anim. Feed Sci. Techno. 78: 21-37. SCHAAFSMA, A. W., L. TAMBURIC-ILLNIC, J. D. MILLER, D. C. HOOKER, 2001. SCHOLLENBERGER, M., H. T. JARA, S. SUEY, W. DROCHNER, H. M. MGLLER, 2002. Fusarium toxins in wheat flour collected in an area in southwest Germany. International Journal of Food Microbiology, 72: 85-89. SCIENTIFIC COMMITTEE ON FOOD, 1999. Opinion on Fusarium Toxins-Part 1: Deoxynivalenol (DON) (expressed on 2 December 1999). Available at http://europa.eu. intlcomm/food/fs/sc/scf/out44_en.pdf STEWART, D. W., L. M. REID, R. W. NICOL and A. W. SCHAAFSMA, 2002. A mathematical simulation of growth of Fusarium in maize ears after artificial inoculation. Phytopathology, 92: 534-541. SUTTON, J. C. 1982. Epidemiology of wheat head blight and maize ear rot caused by Fusarium graminearum. Canadian Journal of Plant Pathology. 4: 195-209. TSCHANZ, A. T., R. K. HORST and P. E. NELSON, 1976. The effect of environment on sexual reproduction of Gibberella zeae. Mycologia 68: 327-340. 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.