Analysis Of Sequences Variance Of Waxy Gene 5' Upstream Regulation Region In Wheat

According to the conserved sequences of Waxy gene 5' upstream regulation region and encoding region,the gene-specific primers were designed to amplify the 5' upstream regulation region from wheat.The PCR amplyfied fragments in Triticum were sequenced and investigated.The results were as follows:1.Four sequences were obtained from Chinese Spring and classified into two types which included two sequences respectively,Typeâ… sequences(about 1100bp)and Typeâ…¡sequences(about 850bp).NCBI Blast indicated that all four sequences were Waxy gene 5' upstream regulation region sequences.Many conserved sequences,SNPs and seventeen deletion regions(more than three bases) were found by further sequencing analysis.All four sequences included regulatory element TATA box located at -1009bp,Typeâ… sequences included regulatory element EB "TGTTATAAG" and GCN4 Motif "AAGTCAAGTC" which located at -667bp and -315bp respectively.2.Nine sequences were obtained from three Chinese Spring Deletants(N4AT4D,N7AT7B and N7DT7B)and classified into two types which included four sequences and five sequences repectively,Typeâ… sequences(about 1100bp) and Typeâ…¡sequences(about 850bp).Typeâ… sequences were not amplyfied from N7DT7B indicated that Typeâ… sequences located at 7D,thus Typeâ…¡sequences located 4A and 7A.NCBI Blast indicated that all nine sequences were Waxy gene 5' upstream regulation region sequences.Many conserved sequences,SNPs and seventeen deletion regions(more than three bases) were found by further sequencing analysis.All nine sequences included regulatory element TATA box located at -1009bp,Typeâ… sequences included regulatory element EB "TGTTATAAG" and GCN4 motif "AAGTCAAGTC" which located at -667bp and -315bp respectively.3.Thirty-four sequences(700-1100bp) were obtained from diploid materials(six sequences were from Triticum monococcum ssp.aegilopoides(AA),Nine sequences were from Triticum monococcum ssp.monococcum(AA),seven sequences were from Triticum Urartu(AA),one sequence were from Ae.speltoides(SS),three sequences were from Ae. longissima(S~1S~1),eight sequences were from Ae.tauschii(DD) ).NCBI Blast indicated that all Thirty-four sequences were Waxy gene 5' upstream regulation region sequences. Many conserved sequences,SNPs and twenty-eight deletion regions(more than three bases) were found by further sequencing analysis.All thirty-four sequences included regulatary element TATA box located at -1011bp,and the sequences from Ae.tauschii(DD)(except the sequences from PI486268) included regulatory element EB"TGTTATAAG" and GCN4 motif"AAGTCAAGTC" which located at -676bp and -316bp respectively.4.Nine sequences(800-900bp) were obtained from tetraploid wheat and classified into tetraploid A type(six sequences) and tetraploid B type(three sequences) with reference to the length of the sequences.NCBI Blast indicated that all nine sequences were Waxy gene 5' upstream regulation region sequences.Many conserved sequences,SNPs and twenty-one deletion regions(more than three bases) were found by further sequencing analysis.Nine sequences included regulatary element TATA box located at -875bp,and tetraploid B type sequences included GCN4 motif "AAGTCAAGTC" which located at -311bp.5.Thirteen sequences were obtained from Triticum aestivum and classified into Typeâ… sequences(about 1100bp),Typeâ…¡sequences(about 850bp) and Yunnan Small(about 900bp) which included seven,five and one sequences repectively.NCBI Blast indicated that all thirteen sequences were Waxy gene 5' upstream regulation region sequences.Many conserved sequences,SNPs and twenty deletion regions(more than three bases) were found by further sequencing analysis.All thirteen sequences included regulatary element TATA box located at -1117bp,and Typeâ… sequences included regulatory element EB "TGTTATAAG" and GCN4 motif "AAGTCAAGTC" which located at -669bp and -315bp respectively.Yunnan Small type included GCN4 motif "AAGTCAAGTC" located at -315bp.6.The clustering results indicated that the sequences from Triticum monococcum ssp. aegilopoides(AA),Triticum monococcum ssp.monococcum(AA),Triticum Urartu(AA) were clusterred together,the sequences from Ae.speltoides(SS) and Ae.longissima(S~1S~1) were clusterred together,the sequences from Ae.tauschii(DD) were clusterred together. Typeâ… sequences and the sequences from Ae.tauschii(DD) were clusterred together.Typeâ…¡sequences,Tetraploid B type sequences and the sequences from Ae.longissima(S~1S~1) were clusterred together,and Yunnan Small type clusterred together with the sequences from Ae.speltoides(SS).The results also indicated that Waxy gene 5' upstream regulation region would be utilized on Triticum evoluation and molecular markers associated selection breeding.