## 2 Materials and Methods
### PCR Analysis  
The PCR reaction was performed using according to GoTaq G2 Green Master Mix protocol (Promega, M7822). The DNA samples were amplified using 11 ISSR markers, as shown in Table 2. The primer's sequences were identified in the literature [37] and they were synthetized at Eurogenetec, Belgium. The PCR mix was prepared in a final volume of 25 \(\upmu\)L. Each reaction was comprised of GoTaq G2 DNA Polymerase, 2X Green GoTaq(r) G2 Reaction Buffer (pH 8.5), 3 mM MgCl\({}_{2}\), 400 \(\upmu\)M each dNTPs, 0.5 \(\upmu\)M primer and 30 ng DNA. Amplifications were performed in an Eppendorf Mastercycler under the following conditions: 94 \({}^{\circ}\)C for 2 min, followed by 30 cycles, each cycle consisting of three steps: (1) 94 \({}^{\circ}\)C for 30 s, (2) 48-58 \({}^{\circ}\)C for 30 s (depending on primer's Tm), (3) 72 \({}^{\circ}\)C for 1 min, and a final step of 72 \({}^{\circ}\)C for 7 min. \begin{table}
\begin{tabular}{c c c c c} \hline
**Genotype Number** & **Scientific Name** & **Accession Name** & **Country** & **Biological Status** \\ \hline TA1 & _T. aestivum_ L. & Izvor & Romania & Modern variety 1 \\ TA2 & _T. aestivum_ L. & Glosa & Romania & Modern variety 1 \\ TA3 & _T. aestivum_ L. & Miranda & Romania & Modern variety 1 \\ TA4 & _T. aestivum_ L. & Andrada & Romania & Modern variety 1 \\ TA5 & _T. aestivum_ L. & Dumbrava & Romania & Modern variety 1 \\ TA6 & _T.asetitium_ L. & Aureluis & Austria & Modern variety 1 \\ TA7 & _T. aestivum_ L. & Softru & France & Modern variety 1 \\ TA8 & _T. aestivum_ L. & Sosthene & France & Modern variety 1 \\ TA9 & _T. aestivum_ L. & Amicus & Austria & Modern variety 1 \\ TA10 & _T. aestivum_ L. & Sothys & France & Modern variety 1 \\ TA11 & _T. aestivum_ L. & Flavor & France & Modern variety 1 \\ TA12 & _T. aestivum_ L. & Solindo & France & Modern variety 1 \\ TA13 & _T. aestivum_ L. & Izalco & France & Modern variety 1 \\ TA14 & _T. aestivum_ L. & Tomage & Austria & Modern variety 1 \\ TA15 & _T. aestivum_ L. & Sophie & France & Modern variety 1 \\ TA16 & _T. aestivum_ L. & Apache & France & Modern variety 1 \\ TA17 & _T. aestivum_ L. & Anapurna & France & Modern variety 1 \\ TA18 & _T. aestivum_ L. & Illico & Switzerland & Modern variety 1 \\ TA19 & _T. aestivum_ L. & Sf. Ilie & Romania & Modern variety 1 \\ TA20 & _T. aestivum_ L. & Lucácesti & Romania & Modern variety 1 \\ TA21 & _T. aestivum_ L. & Udesti 1 & Romania & Modern variety 1 \\ TA22 & _T. aestivum_ L. & Udesti 2 & Romania & Modern variety 1 \\ TA23 & _T. aestivum_ L. & Udesti 3 & Romania & Modern variety 1 \\ TA24 & _T. aestivum_ L. & Frumosa & Romania & Modern variety 1 \\ TA25 & _T. aestivum_ L. & Tsäuti & Romania & Modern variety 1 \\ TM26 & _T. monocytogenes_ L. & SVGB-11842 & Romania & Landrace 2 \\ TM27 & _T.monocoecum_ L. & SVGB-11861 & Romania & Breeding line 3 \\ TS28 & _T. spelta_ L. & Ebners Rotkorn & Austria & Modern variety 1 \\ TS29 & _T. spelta_ L. & Frankenkorn & Austria & Modern variety 1 \\ TS30 & _T. spelta_ L. & Alkoran & Russia & Modern variety 1 \\ TS31 & _T. spelta_ L. & Oberkulmer Rotkorn & Germany & Modern variety 1 \\ \hline \end{tabular}  
* Currently cultivated variety, which is distinguished by superior characteristics of quality, productivity, uniformity and stability compared to a primitive variety and is widely used as a parent in the breeding program. 2 Local variety of a plant species that has been obtained under the action of natural and/or artificial empirical selection, in specific environmental conditions and that presents a series of distinct individual characteristics in order to be associated with a specific geographical region. 3 Biological material obtained by breeders through artificial selection, based on conscious selection schemes.  
\end{table} 
Table 1: Identification of genotypes regarding species, accession name, origin and biological status. some wheat samples of which wet gluten values were less than 22% were difficult to be used by bakery producers [25].