In collaboration with Payame Noor University and Iranian Biotechnology Society

Document Type : Research Paper

Authors

professor assistant

Abstract

Fusarium Head Blight (FHB), which is caused commonly by Fusarium graminearum, has ability to result significant reduce in yield and also cause indirect losses due to the accumulation of potent mycotoxins (trichothecenes) in harvested grain as secondary metabolites; which are hazardous for human and animal. Trichothecene mycotoxins (such as DON) are potent protein synthesis inhibitors for eukaryotic organisms. Trans expression of AYT1 gene from S. cerevisiae capable of trichothecene 3-O-acetylation and converts DON to a less toxic acetylated form. One of the detoxification and resistance to mycotoxins is acetylation. The main goal of this study was to evaluate transgenic tobacco model plants to deoxynivalenol (DON). In order to detect expression of the AYT1 transgene, we added cMyc tag via PCR-Tagging method and introduced it into the model tobacco plants through Agrobacterium-mediated transformation in an attempt to detoxify DON. After confirmation of integrations of AYT1-cMyc into the tobacco genome by molecular analyses, Immuno-blotting and serological protein studies and trichothecene acetyl transferase activity analyses confirmed the expression of AYT1 and tolerance to 10 ppm concentration of DON in transgenic lines was observed.

Keywords

Main Subjects

 
REFERENCES
 
Bai GH, Desjardins AE and Plattner RD (2002) Deoxynivalenol-nonproducing Fusarium graminearum causes initial infection but does not cause disease spread in wheat spikes. Mycopathologia. 153: 91–98.
Chen L, McCormick SP, Hohn TM (2000) Altered Regulation of 15-Acetyldeoxynivalenol Production in Fusarium graminearum. Appl Environ Microbiol. 66(5): 2062–2065.
Eudes F, Comeau A, Rioux S, Collin J (2000) Phytotoxicité de huit mycotoxines associées à la fusariose de l’épi chez le blé. Can. J. Plant Pathol. 22: 286–292.
Kimura M, Kaneko I, Komiyama M, Takatsuki A, Koshino H, Yoneyama K, Yamaguchi I (1998) Trichothecene 3-O-acetyltransferase protects both the producing organism and transformed yeast from related mycotoxins. J Biol Chem 273: 1654–1661.
Manoharan M, Dahleen LS, Hohn TM, Neate SM, Yu X, Alexander NJ, McCormick SP, Bregitzer P, Schwarz PB, Horsley RD (2006) Expression of 3-OH trichothecene acetyltransferase in barley (Hordeum vulgare L.) and effects on deoxynivalenol. Plant Science. 171: 699–706.
Muhitch MJ, McCormick SP, Alexander NJ, Hohn TM (2000) Transgenic expression of the TRI101 or PDR5 gene increases resistance of tobacco to the phytotoxic effects of the trichothecene 4,15- diacetoxyscirpenol. Plant Sci. 157: 201 – 207.
Ohsato Sh , Ochiai-Fukuda T,· Nishiuchi T, Takahashi-Ando N,· Koizumi Sh, Hamamoto H, Kudo T, Yamaguchi I, Kimura M (2007) Transgenic rice plants expressing trichothecene3-O-acetyltransferase show resistance to the Fusarium phytotoxin deoxynivalenol. Plant Cell Rep. 26: 531–538.
Kimura M, Takahashi-Ando N, Nishiuchi T, Ohsato S, Tokai T, Ochiai N, Fujimura M, Kudo T, Hamamoto H, Yamaguchi I (2006) Molecular biology and biotechnology for reduction of Fusarium mycotoxin contamination. Pesticide Biochemistry and Physiology. 86: 117–123.
Alexander NJ, McCormick SP,Hohn TM (2002) The identification of the Saccharomyces cerevisiae gene AYT1(ORF-YLL063c) encoding an acetyltransferase. Yeast (19): 1425-1430.
Hoffman CS, Winston FA (1987) Ten-minute DNA preparation from yeast efficiently releases autonomous plasmid for transformation of Escherichia coli. Gene. 57(2-3): 267-72.
Doyle JJ, Doyle JL (1987) A rapidDNAisolation procedure for small quantities of fresh leaf tissue. Phytochem. Bulletin. 19: 11 – 15.
Bai GH, Shaner G (2004) Management and Resistance in Whaet and Barely to Fusarium Head Blight. Annul. Rev. Phytopathol. 42: 135 – 161.
Abolmaali SH, Mitterbauer R, Adam G (2008) Engineered bakers yeast as a sensitive bioassay indicator organism for the trichothecene toxin deoxynivalenol. J Microbiol Methods. 72(3): 306 – 12.
Ittu M, Hagima I, Moraru I, Raducanu F (1995) Reaction of some wheat and triticale genotypes to toxins, culture filtrates and cultures of Fusarium. In vivo screening and the relation between results obtained in vivo and in vitro. Theor. Appl. Genetics. 27: 1–13.
Bottalico A, Logrieco A, Visconti A (1989) Fusarium species and their mycotoxins in infected corn in Italy. Mycopathologia. 107: 85 – 92.
Snijder CHA (2004) Resistance in wheat to Fusarium infection and trichothecene formation. Toxicology Letters. 153: 37 – 46.