Dumur Jérôme, Jahier Joseph, Bancel Emmanuelle, Laurière Michel, Bernard Michel, Branlard Gérard
UMR INRA-ENSAR, Amélioration des Plantes et Biotechnologies Végétales, Le Rheu, France.
Proteomics. 2004 Sep;4(9):2685-95. doi: 10.1002/pmic.200300800.
Three monosomic lines (MSLs) and three nullisomic lines (NSLs) of the homeologous group 1 and one euploid line of the bread wheat Triticum aestivum cultivar Courtot were used in a proteomic approach to investigate the effects of zero, one or two doses of chromosomes 1A, 1B and 1D on the amount of endosperm proteins. Polypeptides whose amounts changed significantly between each aneuploid line and the euploid line were identified using image analyses of two-dimensional gel electrophoresis patterns resulting from specific endosperm protein extractions. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry and electrospray ionization tandem mass spectrometry were also used for protein identification. Removing one chromosome or a chromosome pair allowed varying responses to be observed for the remaining endosperm protein genes. Compensation phenomena for the high molecular weight glutenin subunits (HMW-GS) were detected only in the MSLs. Subunits Bx7, By8 and Dy12 were the only HMW-GS overexpressed (from 152-737%) when chromosomes 1A or 1B or 1D were at hemizygous state. Thirteen new protein spots were detected only in the NSL1D, and seven were identified as HMW-GS analogs. These seven new spots may result from the expression of inactive genes. The HMW-GS were of significantly higher volume in MSLs, whereas the low molecular weight glutenin subunits and the gamma-gliadins were of lower volume in aneuploid lines. Most of the down-regulated proteins in the MSLs were storage proteins encoded at loci located on another chromosome pair. Complex regulations between chromosomes and loci of the homeologous groups 1 and 6 in bread wheat are discussed.
采用蛋白质组学方法,利用普通小麦品种科托(Triticum aestivum cultivar Courtot)的3条1号部分同源群单体附加系(MSLs)、3条1号部分同源群缺体附加系(NSLs)以及1条整倍体系,研究1A、1B和1D染色体零剂量、单剂量或双剂量对胚乳蛋白含量的影响。通过对特定胚乳蛋白提取物进行二维凝胶电泳图谱的图像分析,鉴定出在每个非整倍体系与整倍体系之间含量有显著变化的多肽。基质辅助激光解吸/电离飞行时间质谱和电喷雾电离串联质谱也用于蛋白质鉴定。去除一条染色体或一对染色体后,可观察到其余胚乳蛋白基因的不同反应。仅在单体附加系中检测到高分子量谷蛋白亚基(HMW-GS)的补偿现象。当1A、1B或1D染色体处于半合子状态时,亚基Bx7、By8和Dy12是仅有的过表达HMW-GS(过表达152%-737%)。仅在缺体附加系1D中检测到13个新的蛋白点,其中7个被鉴定为HMW-GS类似物。这7个新蛋白点可能是由无活性基因的表达产生的。单体附加系中HMW-GS的体积显著更大,而非整倍体系中低分子量谷蛋白亚基和γ-醇溶蛋白的体积更小。单体附加系中大多数下调的蛋白是由位于另一对染色体上的位点编码的贮藏蛋白。本文还讨论了普通小麦1号和6号部分同源群染色体与位点之间的复杂调控。