Studying of Chromosomal Substitution on protein Banding Patterns of high molecular weight-glutenin's (HMW-GS) subunits in wheat MAHMOOD SOLOUKI , ABBASALI EMAMJOMEH * Department of Agronomy and Plant Breeding University of Zabol Mofateh Street  Jahad Square  Zabol IRAN . ac . zabol @ imamjomeh _ a Abstract: - Substitution lines use for chromosomal location of genes. A storage protein of wheat seed contains gliadins and Glutenins. Glutenins is consisted of high molecular weight and low molecular weight. We extracted proteins from 27 substitution lines related to kapla  shayen substitution lines and Chinese Spring as control samples using laemmli et al (1970) protocol. After protein extraction, the samples were electrophoresed using SDS-PAGE. We stained the gels both commassie blue R250 and silver nitrate methods. We scored the bands based on 0 for absence and 1 for presence of each band. Then similarity matrix was calculated using Jaccard coefficient. Payne scoring system was calculated for each line. We observed 11 polymorphic bands. The ranges of coefficients of similarity were 0.55 -1. The coefficients of similarity were 1 for the most of lines. The coefficients of similarity of substitution lines of kapla and parent were 1 but about kapla 3A was 0.736. The coefficient of similarity of kapla 3A and other lines was low. These lines and control were clustered to 3 groups that kapla 3A and shayen 4B were in 2 separate and single groups. Based on silver staining, the range of coefficients of similarity were from 0.7 - 1. The coefficients of similarity of almost all lines were 1. Kapla 3A, kapla 4B and shayen 7B had lowest similarity. According cluster analysis, these lines and control were clustered to 2 groups that kapla 3A was in the separate group. Based on Payne scoring system, only kapla 3A had 2* and 10 bands that can be main reason of difference to other lines and control. According to these scoring the bands of all lines (except kapla 3A) were similar to parent kapla and Chinese Spring .But some of lines of shayen and control were different and these lines were clustered to 3 groups that shayen 7A , shayen 4B and kapla 3A were in the separate group. In the kapla cultivar, 3A substitution chromosome and in the shayen cultivar 7A and 4B chromosomes had high variation in the band pattern. Key- words: - Substitution Lines, Glutenin, HMW-GS, Silver Staining and Commassie Blue Staining. 1 Introduction Substitution lines are produced using chromosomal substitution between two cultivars (without changing in chromosome number) [1]. These lines are produced using aneuploid lines [9]. All differences between two cultivars is related to substituted chromosomes [2].These lines were used for determination of chromosomal location of genes related to environmental stresses [14]. Substitution lines are produced for different cultivars of wheat, because aneuploidy can tolerate by wheat [7]. Different alleles may produce proteins with different combinations of amino acids that can be segregate well using SDS-PAGE method [11]. Gluten (proteins of wheat seed) is effective on bread making quality; therefore determination of chromosomal location of these alleles is useful for genetic improvement of bread making quality of wheat [10]. Determination of glutenin's alleles is used highly in projects of wheat breeding [6]. Studying of storage proteins has been reported in many researches such as phylogenetic relationships, genomic homologies and genetic diversity [8]. The first chromosomal experiment using substitution lines was accomplished in cultivars of tacher and hoop (triticum aestivum). Dashti et al (1998) determined chromosomal location of heading date alleles in bread wheat. They showed that location of these genes are on chromosome 3A, 3D, and 1D in vichita cultivar and on 3A,3D,1D,4D,6A and 1B in shayen cultivar [4]; Then 1A,1D and 6D chromosomes are effective on Proceedings of the 2007 WSEAS Int. Conference on Cellular & Molecular Biology - Biophysics & Bioengineering, Athens, Greece, August 26-28, 2007 78