Real-time PCR and agar plating method to predict Fusarium verticillioides and fumonisin B1 content in Nigerian maize

• Autorzy: Adejumo T O , Hettwer U. , Nutz S. , Karlovsky P.

Warianty tytułu

PL

Real-time PCR i metoda plytkowa do przewidywania zawartosci fumonisinu B1 wytwarzanego przez Fusarium verticillioides w nigeryjskiej kukurydzy

• Języki publikacji: EN

Abstrakty

EN

Eighty maize grain samples collected in Nigeria were investigated for fumonisin B1 (FB1) content and Fusarium verticillioides colonization. F. verticillioides DNA was quantified by species-specific real-time PCR and living propagules of the fungus were counted by agar-plating method. FB1 was detected in 55 (68.7%) of the total samples (mean: 98.5 μg/kg, range: 10 to 714 μg/kg) at 10 μg/kg detection limit. The mean amount of F. verticillioides DNA determined by real-time PCR was 49.7 μg/kg (range: 10-126.7 μg/kg), while agar plate method showed the presence of F. verticillioides in 45 samples (mean incidence: 21.0%, range: 6.7-60.0%). There was correlation ties between F. verticillioides DNA by real time PCR and fungal colonization by agar plate method (R = 0.71, p = 00001 at 95% confidence level), and means of FB1 and F. verticillioides DNA in the yellow and white maize were significantly different. Despite the high consumption of maize in Nigeria, the amount of FB1 ingested by consumers appears to be low. The estimated daily intake of fumonisins was 0.21 μg/kg body weight per day.

PL

Badano 80 prób nasion kukurydzy zebranych w Nigerii na zawartość fumonisinu B1 (FB1) i zasiedlenie ich przez Fusarium verticillioides. Określono zawartość DNA wykorzystując specyficzną dla gatunku metodę real-time PCR i obliczono żywotne elementy rozmnożeniowe grzyba metodą płytkową. FB1 wykryto w 55 (68,7%) przypadkach w ogólnej liczbie prób (średnia: 98,5 μg/kg, zakres: 10 do 714 μg/kg), przy limicie wykrywalności 10 μg/kg. Średnia ilość DNA F. verticillioides określona metodą rzeczywistego czasu PCR wynosiła 49,7 μg/kg (zakres: 10-126,7 μg/kg), podczas gdy metoda płytkowa ujawniła obecność F. verticillioides w 45 próbach (średnie występowanie: 21,0%, zakres 6,7-60,0%). Wystąpiły powiązania korelacyjne między DNA F. verticillioides ujawnione przez real-time PCR i zasiedleniem w przypadku metody płytek agarowych, (R = 0,71, p = 00001 przy poziomie ufności 95%), a średnie dla FB1 oraz DNA F. verticillioides w żółtej i białej kukurydzy były istotnie różne. Pomimo wysokiej konsumpcji kukurydzy w Nigerii ilość FB1 spożywanego przez konsumentów wydaje się niska. Szacunkowe dzienne spożycie fumonisinu wynosiło 0,21 μg/kg w stosunku do wagi ciała.

Słowa kluczowe

PL

Nigeria; maize; fumonisin B1; Fusarium certicillioides; real-time polymerase chain reaction; agar plate method; prediction

• Wydawca: -
• Czasopismo: Journal of Plant Protection Research
• Rocznik: 2009
• Tom: 49
• Numer: 4
• Opis fizyczny: p.399-404,ref.

Twórcy

autor

Adejumo T O

• Adekunle Ajasin University, P.M.B. 01, Akungba-Akoko, Nigeria

autor

Hettwer U.

autor

Nutz S.

autor

Karlovsky P.

Bibliografia

• Adejumo T. O., Hettwer U., Karlovsky P. 2007a. Occurrence of Fusarium species and trichothecenes in Nigerian maize. Int. J. Food Microbiol. 116: 350-7.
• Adejumo T. O., Hettwer U., Karlovsky P. 2007b. Survey of maize from south western Nigeria for Zearalenone, α- and β- Zearalenols, Fumonisin B1, and Enniatins produced by Fusarium species. Food Addit. Contam. 24 (9): 993-1000.
• Bankole S. A., Mabekoje O. O. 2004. Occurrence of aflatoxins and fumonisins in preharvest maize from south-western Nigeria. Food Addit. Contam. (21) 3: 251-255.
• Bottalico A. 1998. Fusarium diseases of cereals: Species complex and related mycotoxin profiles in Europe. J. Plant Pathol. 80: 85-103.
• Brandfass C., Karlovsky P. 2006. Simultaneous detection of Fusarium culmorum and F. graminearum in plant material by duplex PCR with melting curve analysis. BMC Microbiol. 6, p. 4.
• Chu F. S., Li G. Y. 1994. Simultaneous occurrence of fumonisin B1 and other mycotoxins in moldy corn collected from the People's Republic of China in regions with high incidences of esophageal cancer. Appl. Environ. Microbiol. (60) 3: 847-852.
• Clear R. M., Patrick S. K., Turkington T. K., Wallis R. 2002. Effect of dry heat treatment on seed borne Fusarium graminearum and other cereal pathogens. Can. J. Plant Pathol. 24: 489-498.
• De Venter T. V. 2000. Emerging food-borne diseases: a global responsibility. Food Nutrit. Agric. 26: 1-13.
• Demeke T., Clear R. M., Patrick S. K., Gaba D. 2005. Species-specific PCR-based assays for the detection of Fusarium species and a comparison with the whole seed plate method and trichothecene analysis. Int. J. Food Microbiol. 103: 271-284.
• Desjardins A. E. 2006. Fusarium Mycotoxins: Chemistry, Genetics, and Biology. APS, St. Paul, MN, 260 pp.
• Desjardins A. E., Manandhar H. K., Plattner R. D., Manandhar G. G., Poling S. M., Maragos C. M. 2000. Fusarium species from Nepalese rice and production of mycotoxins and gibberellic acid by selected species. Appl. Environ. Microbiol. (66) 3: 1020-1025.
• Donaldson G. C., Ball L. A., Axelrood P. E., Glass N. L. 1995. Primer sets developed to amplify conserved genes from filamentous ascomycetes are useful in differentiating Fusarium species associated with conifers. Appl. Environ. Microbiol. 61 (44): 1331-1340.
• El-Sayed A. M. A., Soher E. A., Sahab A. F. 2003. Occurrence of certain mycotoxins in corn and corn-based products and thermostability of fumonisin B1 during processing. Nahrung/Food (47) 4: 222-225.
• European Commission. 2003. Updated opinion of the Scientific Committee on Food on Fumonisin B1, B2 and B3. Expressed on 4 April 2003. http://europa.eu.int/comm/food/fs/sc/scf/out185_en.pdf
• Gelderblom W. C. A., Kriek N. P. J., Marasas W. F. O., Thiel P. G. 1991. Toxicity and carcinogenicity of the Fusarium moniliforme metabolite, fumonisin B1 in rats. Carcinogenesis 12: 1247-1251.
• Harrison L. R., Colvin B. M., Greene J. T., Newman L. E., Cole J. R. 1990. Pulmonary edema and hydrothorax in swine produced by fumonisin B1, a toxic metabolite of Fusarium moniliforme. J. Vet. Diagn. Invest. 2: 217-221.
• Jurado M., Vaźquez C., Marıń S., Sanchis V., Gonzalez-Jaeń M. T. 2006. PCR-based strategy to detect contamination with mycotoxigenic Fusarium species in maize. Syst. Appl. Microbiol. 29 (8): 681-689.
• Katta S. K., Cagampang A. E., Jackson L. S., Bullerman L. B. 1997. Distribution of Fusarium molds and fumonisins in drymilled corn fractions. Cereal Chem. 74: 858-863.
• Kellerman T. S., Marasas W. F. O., Thiel P. G., Gelderblom W. C. A., Cawood M., Coetzer J. A. W. 1990. Leukoencephalomalacia in two horses induced by oral dosing of fumonisin B1. Onderstepoort J. Vet. Res. 57: 269-275.
• Leslie J. F., Summerell B. A. 2006. The Fusarium Laboratory Manual. Blackwell Publishing, 400 pp.
• Leslie J. F., Pearson C. A., Nelson P. A., Toussoun T. A. 1990. Fusarium spp. from maize, sorghum, and soyabean fields in the central and eastern United States. Phytopathology 86: 343-350.
• Logrieco A., Moretti A., Ritieni A., Chelkowski J., Altomare C., Bottalico A., Randazzo G. 1993. Natural occurrence of beauvericin in preharvest Fusarium subglutinans infected maize ears in Poland. J. Agric. Food Chem. 41: 2149-2152.
• Mule G., Susca A., Stea G., Moretti A. 2004. A species-specific PCR assay based on the calmodulin partial gene for identification of Fusarium verticillioides F. proliferatum and F. subglutinans. Eur. J. Plant Pathol. 110: 495-502.
• Mulfinger S., Niessen L., Vogel R. F. 2000. PCR based quality control of toxigenic Fusarium spp. in brewing malt using ultrasonication for rapid sample preparation. Adv. Food Sci. 22: 38-46.
• Murray M. G., Thompson W. F. 1980. Rapid isolation of high molecular-weight plant DNA. Nucleic Acids Res. 8: 4321-4325.
• Prange A., Modrow H., Hormes J., Kramer J., Kohler P. 2005. Influence of mycotoxin producing fungi (Fusarium, Aspergillus, Penicillium) on gluten proteins during suboptimal storage of wheat after harvest and competitive interactions between field and storage fungi. J. Agric. Food Chem. 53: 6930-6938.
• Rava E., Viljoen J. H., Kallmeyer H., de Jager A. 1996. Fungi and mycotoxins in South African maize of the 1993 crop. Mycotoxin Res. 12: 15-24.
• Rheeder J. P., Marasas W. F. O., Thiel P. G., Sydenham E. W., Shepherd G. S., Van Schalkwyk D. J. 1992. Fusarium moniliforme and fumonisins in corn in relation to esophageal cancer in Transkei. Phytopathology 82: 353-357.
• Royer D., Humpf H. U., Guy P. A. 2004. Quantitative analysis of Fusarium mycotoxins in maize using accelerated solvent extraction before Liquid Chromatography/Atmospheric Pressure Chemical Ionization Tandem Mass Spectrometry. Food Addit. Contam. 21 (7): 678-692.
• Sambrook J., Fritsch E. F., Maniatis T. 1989. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY: 11: 271-282, 18.60-18.75.
• Schnerr H., Niessen L., Vogel R. F. 2001. Real time detection of the tri5 gene in Fusarium species by LightCycler™-PCR using SYBRR Green I for continuous fluorescence monitoring. Int. J. Food Microbiol. 71: 53-61.
• Stewart C. N. 1993. A rapid CTAB DNA isolation technique useful for rapid fingerprinting and other PCR applications. Biotechniques 14, p. 748.
• Strange R. N., Scott P. R. 2006. Plant Disease: a threat to global food security. Ann. Rev. Phytopathol. 43: 83-116.
• Taylor J. W., Geiser D. M., Burt A., Koufopanou V. 1999. The evolutionary biology and population genetics underlying fungal strain typing. Clin. Microbiol. Rev. 12: 126-146.
• Ueno Y., Iijima K., Wang S. D., Sugiura Y., Sekijima M., Tanaka T., Chen C., Yu Sz. 1997. Fumonisins as possible contributory risk factors for primary liver cancer: a 3-year study of corn harvested in Haimen, China by HPLC and ELISA. Food Chem. Toxicol. 35: 1143-1150.
• Viquez O. M., Castell-Perez M. E., Shelby R. A. 1996. Occurrence of fumonisin B1 in maize grown in Costa Rica. J. Agric. Food Chem. 44: 2789-2791.
• Voss K. A., Chamberlain W. J., Bacon C. W., Norred W. P. 1993. A preliminary investigation on renal and hepatic toxicity in rats fed purified fumonisin B1. Nat. Toxins 1: 222-228.
• Waalwijk C., van der Heide R., de Vries I., van der Lee T., Schoen C., Corainville G. C., Hä user-Hahn I., Kastelein P., Kfhl J., Lonnet P., Demarquet T., Kema G. H. J. 2004. Quantitative detection of Fusarium species in wheat using TaqMan. Eur. J. Plant Pathol. 110: 481-494.
• Williams K. C., Blaney B. J., Young R. A., Peters R. T. 1992. Assessment for animal feed of maize kernels naturally infected predominantly with Fusarium moniliforme and Diplodia maydis. II. Nutritive value as assessed by feeding to rats and pigs. Aust. J. Agric. Res. 43: 783-94.