Molecular characterization of gliadins of Chinese Spring wheat in relation to celiac disease elicitors
Molecular characterization of gliadins of Chinese Spring wheat in relation to celiac disease elicitors
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DOI：
10.1266/ggs.17-00034
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发表时间：
2018-02-01
影响因子：
1.1
通讯作者：
Ogihara, Yasunari
中科院分区：
文献类型：
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作者：
Kawaura, Kanako;Miura, Mayuko;Ogihara, Yasunari
小麦种子贮藏蛋白麦胶蛋白和谷蛋白是由多基因编码的。麦胶蛋白进一步分为α -、γ -、δ -和ω -麦胶蛋白。编码α -麦胶蛋白的基因属于一个庞大的多基因家族，其成员位于同源组6染色体上的gli2位点。编码其他麦胶蛋白的基因位于同源组1染色体上的麦胶蛋白1位点上。采用二维聚丙烯酰胺凝胶电泳（2-DE）对麦胶蛋白进行了表征。本研究对中国春小麦非整倍体系的麦胶蛋白进行了比较。2-DE后，麦胶蛋白分离成70个点，在6A、6B和6D染色体上分别编码了10个、10个和16个点，表明它们是α -麦胶蛋白。同样，在1A、1B和1D染色体上分别编码了6、3和7个点，这表明它们是γ -麦胶蛋白。不能分配给染色体的点进行n端测序，并确定为α -麦胶蛋白或γ -麦胶蛋白。2-DE谱显示，分配给6D染色体的特异性α -麦胶蛋白位点在四染色体组2A染色体中丢失。此外，针对腹腔疾病（CD）表位glia - α 9肽的western blotting结果表明，在染色体6D上含有CD表位的α -麦胶蛋白在染色体2A四染色体系中缺失。利用2-DE、定量RT-PCR和基因组PCR对α -麦胶蛋白进行系统分析发现，四染色体2A系在glii - d2位点携带染色体片段的缺失。Gli-D2位点的这种结构改变可能为降低CD免疫毒性的育种计划提供遗传资源。
The wheat seed storage proteins gliadin and glutenin are encoded by multigenes. Gliadins are further classified into alpha-, gamma-, delta- and omega-gliadins. Genes encoding alpha-gliadins belong to a large multigene family, whose members are located on the homoeologous group 6 chromosomes at the Gli-2 loci. Genes encoding other gliadins are located on the homoeologous group 1 chromosomes at the Gli-1 loci. Two-dimensional polyacrylamide gel electrophoresis (2-DE) was used to characterize and profile the gliadins. The gliadins in aneuploid Chinese Spring wheat lines were then compared in this study. Gliadin proteins separated into 70 spots after 2-DE and a total of 10, 10 and 16 spots were encoded on chromosomes 6A, 6B and 6D, respectively, which suggested that they were alpha-gliadins. Similarly, six, three and seven spots were encoded on chromosomes 1A, 1B and 1D, respectively, which indicated that they were gamma-gliadins. Spots that could not be assigned to chromosomes were N-terminally sequenced and were all determined to be alpha-gliadins or gamma-gliadins. The 2-DE profiles showed that specific alpha-gliadin spots assigned to chromosome 6D were lost in tetrasomic chromosome 2A lines. Furthermore, western blotting against the Glia-alpha 9 peptide, an epitope for celiac disease (CD), suggested that alpha-gliadins harboring the CD epitope on chromosome 6D were absent in the tetrasomic chromosome 2A lines. Systematic analysis of alpha-gliadins using 2-DE, quantitative RT-PCR and genomic PCR revealed that tetrasomic 2A lines carry deletion of a chromosome segment at the Gli-D2 locus. This structural alteration at the Gli-D2 locus may provide a genetic resource in breeding programs for the reduction of CD immunotoxicity.