Analysis and validation of genome-specific DNA variations in 5′flanking conserved sequences of wheat low-molecular-weight glutenin subunit genes  

作　　者：BAUM Bernard NEVO Eviatar 

机构地区：[1]Agriculture & Agri-Food Canada, Eastern Cereal and Oilseed Research Centre, Ottawa K1A 0C6, Canada [2]Institute of Evolution, University of Haifa, Mt. Carmel, Haifa 31905, Israel

出　　处：《Science China(Life Sciences)》2006年第4期322-331,共10页中国科学（生命科学英文版）

基　　金：supported by the National High Technology Research and Development Program of China(863 Program)(Grant No.2003AA207100);National Natural Science Foundation of China(Grant Nos.30300219&30571163);the Foundation for the Author of National Excellent Doctoral Dissertation of China from Ministry of Education,China(Grant Nos.200357&200458);supported by the Program for Changjiang Scholars and Innovative Research Teams in University of China(Grant No.IRT0453)

摘　　要：The thirty-three 5′ flanking conserved sequences of the known low-molecular-weight subunit (LMW-GS) genes have been divided into eight clusters, which was in agreement with the classification based on the deduced N-terminal protein sequences. The DNA polymorphism between the eight clusters was obtained by sequence alignment,; a total of 34 polymorphic positions were observed in the approximately 200 bp regions, among which 18 polymorphic positions were candidate SNPs. Seven cluster-specific primer sets were designed for seven out of eight clusters containing cluster-specific bases, with which the genomic DNA of the ditelosomic lines of group 1 chromosomes of a wheat variety ‘Chinese Spring’ was employed to carry out chromosome assignment. The subsequent cloning; DNA sequencing of PCR fragments validated the sequences specificity of the 5′ flanking conserved sequences between LMW-GS gene groups in different genomes. These results suggested that the coding; 5′ flanking regions of LMW-GS genes are likely to have evolved in a concerted fashion. The seven primer sets developed in this study could be used to isolate the complete ORFs of seven groups of LMW-GS genes, respectively,; therefore possess great value for further research in the contributions of a single LMW-GS gene to wheat quality in the complex genetic background; the efficient selections of quality-related components in breeding programs.The thirty-three 5′ flanking conserved sequences of the known low-molecular-weight subunit (LMW-GS) genes have been divided into eight clusters, which was in agreement with the classification based on the deduced N-terminal protein sequences. The DNA polymorphism between the eight clusters was obtained by sequence alignment, and a total of 34 polymorphic positions were observed in the approximately 200 bp regions, among which 18 polymorphic positions were candidate SNPs. Seven cluster-specific primer sets were designed for seven out of eight clusters containing cluster-specific bases, with which the genomic DNA of the ditelosomic lines of group 1 chromosomes of a wheat variety ‘Chinese Spring’ was employed to carry out chromosome assignment. The subse-quent cloning and DNA sequencing of PCR fragments validated the sequences specificity of the 5′ flanking conserved sequences between LMW-GS gene groups in different genomes. These results suggested that the coding and 5′ flanking regions of LMW-GS genes are likely to have evolved in a concerted fashion. The seven primer sets developed in this study could be used to isolate the com-plete ORFs of seven groups of LMW-GS genes, respectively, and therefore possess great value for further research in the contributions of a single LMW-GS gene to wheat quality in the complex genetic background and the efficient selections of quality-related components in breeding programs.

关 键 词：wheat  LMW-GS gene  5' flanking sequence  DNA polymorphism  SNP  GENE cloning. 

分 类 号：S512.1[农业科学—作物学] Q943[生物学—植物学]