Szeged - Timisoara axis for the safe food and feed HURO / 0901/147/2.2.2 Acronym: SZETISA 1 LP - Szeged University PP- Banat University of Agricultural Sciences and Veterinary Medicine, Timisoara
Determination of microelements as affected by microbial toxins in wheat Fusarium detection The analyzed material Artificially infected material - 2 wheat cultivars - Szeged Fusarium graminearum Toxin quantification Fusarium quantification SYBR Green Different organs Different periods of infection Fc Fc Fusarium culmorum Fg Fg 15A2
Wheat specimen spontaneously and naturally infected - collected in Romania Samples – 2010 – 6 samples collected from diferent regions Samples -– the samples will be collected from large wheat farms from Timis and Arad county (July, 2011) Fusarium strain identification Fusarium graminearum, Fusarium culmorum – producing deoxinivalenol Fusarium verticillioides - producing fumonizin Fusarium proliferatum - producing fumonizin Toxin quantification How many samples?
The analyzed material Determination of microelements as affected by microbial toxins in maize. Samples – 2010 – 2 samples Bt and conventional feed containing soybean and corn Samples – the samples will be collected from farms where GMO corn is cultivated and also from large corn farms from Timis and Arad county (September, 2011) 2 corn cultivars - genetically modified (Bt) and conventional from different locations Genetically modification confirmation Fusarium strain identification Toxin quantification How many samples?
DNA extraction and purification WORKING METHODOLOGY Fungi Fungi from grains - wheat and corn feed PCR amplification Toxin quantification Strain identification Gene encoding the toxins Quantitative PCR Strain quantification Identifications of Fusarium strains toxin producers and nonproducers – distinguished on the basis of DNA polymorphism in the –tubulin gene as well as in the large ribosomal subunit or the internal transcribed spacer
Transgene detection MAIZE SAMPLES Mon 810 construct gene
Determination of microelements as affected by heavy metal load in alfalfa. Two alfalfa cultivars of different susceptibilities to heavy metal (HM) were selected. The analyzed material Satelit (sensitive) Sigma (tolerant)
Heavy metals treatments (Cd, Pb, Cu, Fe) DNA fingerprint Heavy metalConc (ppm) Cd Pb Cu Fe???? Preparing alfalfa plants for all the experimental variants – growth chamber The genetic relationship between two genotypes The intragenotype characterization
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