title: PCR-based isolation and identification of full-length low-molecular-weight glutenin subunit genes in bread wheat (Triticum aestivum L.).
abstract: Low-molecular-weight glutenin subunits (LMW-GSs) are encoded by a multi-gene family and are essential for determining the quality of wheat flour products, such as bread and noodles. However, the exact role or contribution of individual LMW-GS genes to wheat quality remains unclear. This is, at least in part, due to the difficulty in characterizing complete sequences of all LMW-GS gene family members in bread wheat. To identify full-length LMW-GS genes, a polymerase chain reaction (PCR)-based method was established, consisting of newly designed conserved primers and the previously developed LMW-GS gene molecular marker system. Using the PCR-based method, 17 LMW-GS genes were identified and characterized in Xiaoyan 54, of which 12 contained full-length sequences. Sequence alignments showed that 13 LMW-GS genes were identical to those found in Xiaoyan 54 using the genomic DNA library screening, and the other four full-length LMW-GS genes were first isolated from Xiaoyan 54. In Chinese Spring, 16 unique LMW-GS genes were isolated, and 13 of them contained full-length coding sequences. Additionally, 16 and 17 LMW-GS genes in Dongnong 101 and Lvhan 328 (chosen from the micro-core collections of Chinese germplasm), respectively, were also identified. Sequence alignments revealed that at least 15 LMW-GS genes were common in the four wheat varieties, and allelic variants of each gene shared high sequence identities (>95%) but exhibited length polymorphism in repetitive regions. This study provides a PCR-based method for efficiently identifying LMW-GS genes in bread wheat, which will improve the characterization of complex members of the LMW-GS gene family and facilitate the understanding of their contributions to wheat quality.
authors.affiliation: State Key Laboratory of Plant Cell and Chromosome Engineering, National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 1 West Beichen Road, Chaoyang District, Beijing 100101, China.
authors.forename: Xiaofei
authors.initials: X
authors.lastname: Zhang
authors.forename: Dongcheng
authors.initials: D
authors.lastname: Liu
authors.forename: Wei
authors.initials: W
authors.lastname: Jiang
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Guo
authors.forename: Wenlong
authors.initials: W
authors.lastname: Yang
authors.forename: Jiazhu
authors.initials: J
authors.lastname: Sun
authors.forename: Hongqing
authors.initials: H
authors.lastname: Ling
authors.forename: Aimin
authors.initials: A
authors.lastname: Zhang
mesh_terms: D001483:Base Sequence; D001939:Bread; D002681:China; D017124:Conserved Sequence / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004587:Electrophoresis, Agar Gel; D017343:Genes, Plant / Q000235:genetics*; D005983:Glutens / Q000737:chemistry* / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D021122:Protein Subunits / Q000737:chemistry* / Q000235:genetics*; D015203:Reproducibility of Results; D016415:Sequence Alignment; D014908:Triticum / Q000235:genetics*
chemical_list: D017931:DNA Primers; D021122:Protein Subunits; D005983:Glutens; C037302:glutenin