A Dissertation Submitted To The Graduate School Of Henan University In Partial Fulfillment Of The Requirements For The Degree Of Master Of Science

Grain hardness is one of the main quality characters of wheat. Puroindoline a(Pina) and Puroindoline b(Pinb) are the major genes controlling grain hardness in wheat.Puroindoline a (Pina) and Puroindoline b (Pinb) play critical roles in determining wheat grain texture, and mutation or absence of either genes results in different grain hardness. There is no large-scale investigation of Ae.tauschii in the Henan, Shaanxi Province of Yellow River and Xinjiang Province where many kinds of Ae.tauschii accessions have been found.In this study,we had analysed the puroindoline protein and alleles using SDS-PAGE and PCR with specific primers.The main content and methods is as follow:(1)Analysis of Puroindoline protein using SDS-PAGE:Puroindoline protein was extracted from the seeds of 51 Ae.tauschii accessions and were separated using SDS-PAGE.SDS-PAGE analysis using seed protein extracts revealed the presence of the putative puroindoline protein whose molecular weight is about 15kD in each of the 51 Ae.tauschii accessions(2)Isolation and characterization of novel variant of puroindoline genes:Based on the reported homologues of Pina and Pinb, two pair of specific primers was designed, and Pina and Pinb was PCR-amplified from 51 accessions belonging to Aegilops genus in the optimum pcr reaction mixture.32 positive PCR products of Pina and 35 positive PCR products of Pinb were sequenced and aligned with those already submitted in GenBank. The results revealed that 30 Pina and 32 Pinb sequences were identical with those in GenBank but from different acessions, while 2 Pina and 3 Pinb sequences were mutated. The 2 mutated Pina and 3 mutated Pinb 447bp in full length, shared a highest identity, and they were deduced to encode 149 amino acid residues with the tryptophan-rich domain (WRW-WKWWK) and (WPTKWWK) which are unique domain in the cereal...