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/ () / / (b a ) * (/ : / : ).. b a..... / /. Pinb-D1e. Pinb-D1c... :.() (Matrix)..( ). (Triticum aestivum).(). fshahria@yahoo.com.au : : *

/ () / /.() () b a..( ) Pinb-D1d Pinb-D1c Pinb-D1e ().( )...( ). (Landrace). (Single kernel characterization system (SKCS))..()..() (Ha).() D a (Pina) a.() (Pinb) b.( ) (Friabilin).().().() ( kda ) () (Domain).() b a.() b (Pina-D1a/Pinb-D1a). b a

... (b a ). Pinb PCR / Pinb PCR. C Pina Pina- Pinb No (Pina null) D1b ().() JMP4. t ( ).. ((α = / -/ -/ )..().. DNA DNA () CTAB DNA.. () Pinb Pina ( ).. (PCR) Touch Down Biometra (T gradient) DNA / DNA Taq dntps / MgCl2. () C C C / C C C. Pinb-D1c Pinb-D1e (Cleaved amplified polymorphic sequences (CAPS)) Pinb-D1c MvaI Pinb-D1e PvuII

/ () / /. ( ) ( C) (bp) Pinb ATG AAG ACC TTA TTC CTC CTA TCA CCA GTA ATA GCC ACT AGG GAA Pina ATG AAG GCC CTC TTC CTCA TCA CCA GTA ATA GCC AAT AGTG ATG AAG ACC TTA TTC CTC CTA CTC ATG CTC ACA GCC GCT No ATG AAG ACC TTA TTC CTC CTA CTC ATG CTC ACA GCC GCC Pina PCR :. : ( - - () - - - - - () - :) Pinb PCR PCR.( - - - () - :) No.(bp DNA Ladder Plus) DNA :M. Pinb Pinb-D1c..() CCG CTG PvuII. Pinb Pinb PCR..( )... Pinb-D1c Pinb-D1e TGG Pinb-D1e.() TGA MvaI

... (b a ). SKCS () () () () Pinb-D1e. (CIMMYT).(). Pinb- ( ). Pinb-D1c D1e ) ( ( ).

/ () / / PvuII Pinb PCR :. Pinb-D1c :.( - - - - - () - - -) Pinb-D1c Pinb-D1e MvaI Pinb PCR : M.(() - - - - - -) Pinb-D1e.(bp DNA Ladder Plus) DNA (P. /). ( ). ( ). D1b b.( ) a a. a.( ) ( ).(P /) ( )...() Pina- Pinb-. Pinb-D1e Pinb-D1c D1b ( ) ( )

... (b a )..... Ha () D....( ).....() ا....... 3. Blochet, J.E., C. Chevalier, E. Forest, E. Pebay-Peyroula, M.F. Gautier, P. Joudrier, M. Pezolet and D. Marion. 1993. Complete amino acid sequence of puroindoline, a new basic and cystine-rich protein with a unique tryptophan-rich domain, isolated from wheat endosperm by Triton X-114 phase partitioning. FEBS Lett. 329: 336-340. 4. Brennan, C.S., B.D. Sulaiman, J.D. Schofield and J.G. Vaughan. 1993. The immunolocation of friabilin and its association with endosperm texture. Asp. Appl. Biol. 36: 69-73. 5. Capparelli, R., G. Borriello, M.J. Giroux and M.G. Amoroso. 2003. Puroindoline A-gene expression is involved in association of puroindolines to starch. Theor. Appl. Genet. 107: 1463-1468. 6. Chen, F., Z.H. He, X.C. Xia, L.Q.Xia, X.Y. Zhang, M. Lillemo and C.F. Morris. 2006. Molecular and biochemical characterization of puroindoline a and b alleles in Chinese landraces and historical cultivars. Theor. Appl. Genet. 112: 400-409. 7. Chen, F., Y. Yu, X. Xia and Z. He. 2007. Prevalence of a novel puroindoline b allele in Yunnan endemic wheats (Triticum aestivum ssp. yunnanense King). Euphytica 156: 39-46. 8. Corona, V., L., Gazza, G. Boggini and N.E. Pogna. 2001. Variation in friabilin composition as determined by A- PAGE fractionation and PCR amplification, and its relationship to grain hardness in bread wheat. J. Cereal Sci. 34: 243-250. 9. Curtis, B.C. 2002. Wheat in the World. published online at: http://www.fao.org/docrep/006/ Y4011E/y4011e04.htm 10. Darlington, H.F., L. Tesci, N. Harris, D. Griggs, I. Cantrell and P.R. Shewry. 2000. Starch granule associated proteins in barley and wheat. J. Cereal Sci. 32: 21-29. 11. Gautier, M.F., M.E. Aleman, A. Guirao, D. Marion and P. Joudrier. 1994. Triticum aestivum puroindolines, two basic cysteine-rich seed proteins: cdna sequence analysis and developmental gene expression. Plant Mol. Biol. 25: 43-57. 12. Gazza, L., F. Nocente, P.K.W. Ng and N.E. Pogna. 2005. Genetic and biochemical analysis of common wheat cultivars lacking puroindoline a. Theor. Appl. Genet. 110: 470-478. 13. Gazza, L., F. Taddei, M. Corbellini, P. Cacciatori and N.E. Pogna. 2008. Genetic and environmental factors affecting grain texture in common wheat. J. Cereal Sci. 47: 52-58. 14. Giroux, M.J. and C.F. Morris. 1997. A glycine to serine change in puroindoline b is associated with wheat grain hardness and low levels of starch-surface friabilin. Theor. Appl. Genet. 95: 857-864. 15. Giroux, M.J. and C.F. Morris. 1998. Wheat grain hardness results from highly conserved mutations in the friabilin..

/ () / / components puroindoline a and b. Proc. Natl. Acad. Sci. USA 95: 6262-6266. 16. Giroux, M.J., L. Talbert, D.K. Habernicht, S. Lanning, A. Hemphill and J.M. Martin. 2000. Association of puroindoline sequence type and grain hardness in hard red spring wheat. Crop Sci. 40: 370-374. 17. Greenwell, P. and J.D. Schofield. 1986. A starch granule protein associated with endosperm softness in wheat. Cereal Chem. 63: 379-380. 18. Lillemo, M. and C.F. Morris. 2000. A leucine to proline mutation in puroindoline b is frequently present in hard wheats from Northern Europe. Theor. Appl. Genet. 100: 1100-1107. 19. Lillemo, M., F. Chen, X. Xia, M. William, R.J. Peňa, R. Trethowan and Z. He. 2006. Puroindoline grain hardness alleles in CIMMYT bread wheat germplasm. J. Cereal Sci. 44: 86-92. 20. Marion, D., B. Bakan and K. Elmorjani. 2007. Plant lipid binding proteins: Properties and applications. Biotechnol. Adv. 25: 195-197. 21. Martin, J.M., R.C. Frohberg, C.F. Morris, L.E. Talbert and M.J. Giroux. 2001. Milling and bread baking traits associated with puroindoline sequence type in hard red spring wheat. Crop Sci. 41: 228-234. 22. Morris, C.F., M. Lillemo, M.C. Simeone, M.J. Giroux, S.L. Babb and K.K. Kidwell. 2001. Prevalence of puroindoline grain hardness genotypes among historically significant North American spring and winter wheats. Crop Sci. 41: 218-228. 23. Morris, C.F. 2002. Puroindolines: the molecular genetic basis of wheat grain hardness. Plant Mol. Biol. 48: 633-647. 24. Morris, C.F. and A.N. Massa. 2003. Puroindoline genotype of the U.S. national institute of standards and technology reference material 8441, wheat hardness. Cereal Chem. 80: 674-678. 25. Pickering, P.A. and M. Bhave. 2007. Comprehensive analysis of Australian hard wheat cultivars shows limited puroindoline allele diversity. Plant Sci. 172: 371-379. 26. Ram, S., N. Jain, J. Shoran and R. Singh. 2005. New frame shift mutation in puroindoline b in Indian wheat cultivars Hyb65 and NI5439. J. Plant Biochem. Biotechnol. 14: 45-48. 27. Saghai-Maroof, M.A., K.M. Soliman, R.A. Jorgensen and R.W. Allad. 1984. DNA spacerlength polymorphism in barley: Mendelian inheritance, Chromosomal location and Population dynamics. Proc. Natl. Acad. Sci. USA. 81: 8014-8018. 28. Turnbull, K.M. and S. Rahman. 2002. Endosperm texture in wheat. J. Cereal Sci. 36: 327-337. 29. Turnbull, K.M., D. Marion, T. Gaborit, R. Appels and S. Rahman. 2003. Early expression of grain hardness in the developing wheat endosperm. Planta 216: 699-706. 30. Turner, M., Y. Mukai, P. Leroy, B. Charef, R. Appels and S. Rahman. 1999. The Ha locus of wheat: identification of a polymorphic region for tracing grain hardness in crosses. Genome 42: 1242-1250.