Tishreen University Journal for Research and Scientific Studies - Biological Sciences Series Vol. (30) No. (1) 2008

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1 Tishreen University Journal for Research and Scientific Studies - iological Sciences Series Vol. (30) No. () 2008 SDS-PGE (UPGM) Jaccard (908 HMW-GS high molecular wheight glutenin subunits HMW-GS

2 Tishreen University Journal for Research and Scientific Studies - iological Sciences Series Vol. (30) No. () 2008 Use of iochemical markers as an effevtive tools to detect genetic variability within tetraploides and hexaploides wheat varieties Dr. Mohammad Mouala Dr. Nizar Mir li** Dr. bdul Rahman Kalhout*** Suha shtar**** (Received 7 / 9 / ccepted 22// ) STRCT In this study two techniques (&SDS-PGE) have been applied to characterize 38 durum wheat varieties and 2 bread wheat varieties. Number of bands resulted by applying the first metohd () was 3 polymorphic bands, while 2 HMW- GS (high molecular wheigt glutenin subunits) have been resulted by applying SDS- PGE. Data were combined together (25 polymorphic bands) to calculate the genetic similarities between studied individuals using Jaccard's coeficient followed by setting up the cluster analysis using UPGM method. The value of genetic diversity was 56% between durum wheat varieties, while the same value was increased to reach 00% in the bread wheat varieties. Dendogram for all 50 genotypes showed a separation of bread wheat varieties in a sub cluster which is distinct from the sub cluster containig the bread wheat varieties. Finally we could conclude the effectivness of seed storage protein marker as a useful tools to characterize individuals, evaluate genetic diversity, registration of new varieties and pedigree analysis studies. Key words: Glutenin, Gliadin, durum wheat, bread wheat, HMW-GS, genetic diversity Prof, Department Of Field Crops, Faculty Of griculture, Tishreen University, Lattakia, Syria. **Head Of Department Of iotechnology, Commission Of tomic Energy. ***Scientist, General Commission For Scientific griculture Research. ****PHd student, Department Of Field Crops, Faculty Of griculture, Tishreen University, Lattakia, Syria.

3 Tishreen University Journal. io. Sciences Series Triticum Spp. (Duvick, 984) Fahima et al.,999, Menkir et al.,997 Ellis,984 (Smith.,984 (Hatzopoulos et al. 2002) ( Shuaib et al., 2007; Mir li 2002 a,b; Redaelli et al., 997; Payne et al., 984) (Mirli, 2000) Metakovsky & ranlard,998 Osborne N Spencer 984 Payne et al., 984; Mir li, 2000 ietz and Wall.,972 ٢٠٣

4 High molecular wheight sub units Low molecular wheight sub units reducing agent LMW-GS HMW-GS HMW-GS (LMW-GS) Gli- D Gli-2 6D 6 6 Pogna et al. (993) Metakovsky et al.(998) HMW-GS LMW Glu- D Pogna et al. (990) Gli- (linked) Glu-3 gel acidic polyacrylamide SDS-poyacrylamide gel SDS-PGE electrophoresis electrophoresis Triticum Durum C

5 Tishreen University Journal. io. Sciences Series Triticum estivum 2 ushuk and Zillman 978) Vortex iorad 50 Rm Relative mobility ushuk & Zillman (978) Payne et al. (98) 2% SDS Laemmli SDS-PGE Tris-HCl (ph 6.8) ٢٠٥

6 Glutenin,Gliadin Numerical Taxonomy and Multivariate nalysis System Jaccard genetic similarity Jaccard, 908) GS(ij)=-[a/(a+b+c)] c b a i j NTSYS Rohlf, 992) coefficient GS(ij) : Cluster analysis (UPGM) RM RM RM RM RM β α unique

7 Tishreen University Journal. io. Sciences Series RM Referen ce ٢٠٧

8 ,27 C 0 2 C 3

9 Tishreen University Journal. io. Sciences Series high molecular low molecular weight SDS-PGE Glutenin subunit wheight HMW-GS C H C ٢٠٩

10 H C

11 Tishreen University Journal. io. Sciences Series SDS-PGE Phylogenetic Jaccard 9 ٢١١

12 Jaccard SDS-PGE Phylogenetic 0 SDS-PGE Phylogenetic Jaccard

13 Tishreen University Journal. io. Sciences Series,, ( Triticum durum Desf Triticum aestivum L. Genome D) (Mechan et al.,978; payne et al.,98a; payne987; Gupta and Shephered 990a) β Gli- Gene silencing Mirli et al.,999 a,b; Payne et al.,98, 984 α Pogna et al.,993 Sozinov and Poperelya (980) Mechan et al., (978) loci). SDS-PGE HMW HMW-GS Johansson et al., (993), Margiotta et al., (996) ( Gene silencing) Mirli., et al., (999a) HMW-GS Mir li., et al., (999b) HMW-GS D D D Mirli., 987 ٢١٣

14 Patterns C SDS-PGE Wei et al., 2000 Fufa et al., 2005; Shuaib et al.,2007 aenziger et al.,200 qualitative trait Mirli (2000)

15 Tishreen University Journal. io. Sciences Series C land race Mir li (2000) D DN 3-ENZIGER, P. S; SHELTON, D.R; SHIPMN M.J. and GRYOSCH. reeding for end-use quality: Reflection on the Nebraska experience. Euphytica,. 200, 9: IETZ, J..; and WLL, J. S. wheat gluten subunits:molecular weights determined by sodium sulfate-polyacrylamide gel electrophoresis. Cereal Chem. 972, 49: ٢١٥

16 5-USHUK, W. and ZILLMN, R.R. Wheat cultivar identification by gliadin electrophoregrams. I. pparatus, method, and nomenclature. Canadian Journal of Plant Science, 978, 58, DUVICK, D.N. Genetic diversity in major farm crops on the farm and in reverse. Economic otany,984, 38, Ellis, J.R.S., The cereal grain trade in the united kingdom the problem of cereal vaiety. Phil. Trans. R. Soc. Lond. V, 984, 304: Fahima, T; Sun, G.L; eharay,.; Krugman, T; eiles,.; Nevo, E. RPD Polymorphism of wild emmer wheat populations, Triticum dicoccoides. Theoritical and applied genetic, 999, 98:v Fufa, H; aenziger, P. S; eecher, I; Dweikat, V; Graybosh, R..; Eskridge K. M.. Comparison of phenotypic and molecular marker-based classification of hard red winter wheat cultivars. Euphytica, 2005, 45: GUPT, R. ; and SHEPHERED, K. W. Two-step one-dimensional SDS-PGE analysis of LMW subunits of glutenin. I. variation and genetic control of the subunits in hexaploid wheats. Theo. pp. Genet, 990a, 80: HTZOPOULOS, P; NILS G; GINNOULI K; GZIS F; NIKOLOUDKIS N; MILIONI D; and HRLMPIDIS K. reeding, molecular markers and molecular biology of the olive tree. Eur J Lipid Sci Technol, 2002, 04: JCCRD, P.. Nouvelles recherches sur la distrubition florale. ull Soc Vaud Sci Nat 44, 908, JOHNSSON, E; HENRIKSSON, P; SVENSSON, G; and HENEEN, W.K. Detection, chromosomal location and evaluation of the functional value of a novel high M r glutenin subunit found in swedish wheats. J. cereal Sci. 7, 993, LEMMLI, U.K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 227, 970, MRGIOTT, ; URNO, M.; COLPRICO, G; JOHNSSON, E; UONOCORE, F; D'OVIDIO, R; and LFINDR, D. Detection of Y type subunit at the Glu- locus in some Swedish bread wheat lines J. Cereal Sci. 23, 996, MECHN, D. K; KSRD, D. D; and QULSET, C.O. Genetic aspects of wheat giadin proteins. iochem. Genet. 6, 978, MENKIR, ; GOLDSROUGH, P; and EJET, G. RPD based assessment of genetic diversity in cultivated races of Sorghum. Crop Sciences, 37, 997, METKOVSKY, E.V and RNLND, G. Genetic diversity of French common wheat germplasm based on gliadin allelles. Theoritical and pplied genetics. 96, 998, MIR LI, N. Protein improvement in Triticum Turgidum Var, durum (DESF.) bu induced mutations and hybridization with Triticum Tugidum Var. dicoccoides (Korn). Ph.D Thesis. The University of New Castle, U.K. 987, MIRLI, N.,. Heterogenity within old and modern durum and bread wheat grown in Syria using the and SDS-PE electrophoresis techniques. Plant varieties & seeds, 3, 2000, MIRLI, N., RI, M.I.E. ND L-SFDI,. High molecular weight glutenin subunits composiyion of Syrian-grown bread wheat and its relationships with gluten strength. J Genetics & reeding, 53, 999a, MIRLI, N; RI, M.I.E. and L-SFDI,. Frequencies of high and low molecular weight gluteninsubunits in durum wheat grown in Syria. Cereal Research Communications. 27, 999b,

17 Tishreen University Journal. io. Sciences Series 23- MIRLI, N. Gliadins composition and cluster analyses of Syrian grown durum wheat. Plant reeding & Seed Science. 46, 2002a, MIRLI, N. Cluster analysis of Syrian grown bread wheat genotypes based on gliadin composition. J.Genet.& reed. 56, 2002b, OSORNE, T.. The proteins of the wheat kernel. Carnegie Inst., Wash. Publ. No. 84, REDELLI, R. P. K;. NG W; POGN N. E. llelic variation at the storage protein loci of 55 US-Grown white wheats. Plant breeding. 6, 997, ROHLF, F. J. NTSYS-PC: numerical taxonomy andmultivariate analysis system version.70.state University of New York, Stony rook N. Y., US., PYNE, P.I; CORFIELD, K.G;, HOLT, L.M. and LCKMN, J... Correlations between the inheritance of certain high molecular weight subunits of glutenin and bread making quality in progenies of six crosses of bread wheat. Journal of Science of Food and griculture 32, 98, PYNE, P. L; HOLT, L. M; JCKSON, E. ;. LW, C. N. Wheat storage proteins:their genetics and their potential for manipulation by plant breeding. Phil. Trans. R. Soc. Lond.. 304, 984, PYNE, P. L; NIGHTINGLE, M. ; KRTTIGER,. F and HOLT, L. M. The relationship between HMW glutenin composition and the breamaking quality of ritish grown wheat varieties. J. Sci. Food gri. 40, 987, POGN, N.E; UTRN, J.C; MELLINI, F; LFINDR, D. and FEILLET, P. Chromosome -encoded gliadins and glutenin subunits in durum wheat: genetics and relationship to gluten strength. Journal of Cereal Science., 990, POGN, P. E; METKOVSKY E. V; REDELLI. R;. RINER F; DCHKEVITCH T. Recombination mapping of Gli-5, a new gliadin-coding locus on chromosomes and in common wheat. Theor.and ppl. Genet.87, 993, SPENCER, D. and OULTER, D. The Physiological Role of Storage Proteins in Seeds. Philosophical Transactions of the Royal Society of London. Series, iological Sciences,. 304,, No. 20, Seed Storage Proteins. 984, SHUI, M; ZE, ; LI, Z; LI, W; HMD, T and KHN, I. Characterization of wheat varieties by seed storage protein electrophoresis. frican Journal of iotechnology. 6 (5), 2007, SMITH, J, S. C; Genetic variability within U.S hybrid maize: multivariate analysis of isozyme data. Crop science. 24, 984, SOZINOV, and POPERELY, F,. Genetic classification of prolamines and its use for plant breeding. nn Tech gric. 29, 980, WEI, YM; ZHENG, YL; LIU, DC; ZHOU, YH; LN, XJ. Gliadin and HMWglutenin variations in Triticum turgidum L. ssp. Turgidum and T. aestivum L. Landraces native to Sichuan, China. Wheat information service. 90, 2000, ٢١٧