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利用春小麦品种 Chinese Spring 的缺失系对小麦胚乳白蛋白和球蛋白进行蛋白质组学和遗传学分析。

Proteomic and genetic analysis of wheat endosperm albumins and globulins using deletion lines of cultivar Chinese Spring.

机构信息

INRA, UMR 1095 GDEC-UBP, 5 chemin de Beaulieu, 63100, Clermont Ferrand, France.

出版信息

Theor Appl Genet. 2012 Nov;125(7):1433-48. doi: 10.1007/s00122-012-1924-5. Epub 2012 Jun 30.

Abstract

Albumins and globulins from the endosperm of Triticum aestivum L. cv Chinese Spring (CS) were analysed to establish a proteome reference map for this standard wheat cultivar. Approximately, 1,145 Coomassie-stained spots were detected by two-dimensional gel electrophoresis (2DE), 410 of which were identified using mass spectrometry and data mining. Salt-soluble endosperm proteins from 67 CS deletion lines were also separated by 2DE (four gels per line). Image analysis of the 268 2DE gels as compared to the CS reference proteome allowed the detection of qualitative and quantitative variations in endosperm proteins due to chromosomal deletions. This differential analysis of spots allowed structural or regulatory genes, encoding 211 proteins, to be located on segments of the 21 wheat chromosomes. In addition, variance analysis of quantitative variations in spot volume showed that the expression of 391 proteins is controlled by one or more chromosome bins with 262 significant increases and 196 significant decreases in spot volume. The spot volume of several proteins was increased or decreased by numerous chromosomal regions and homoeologous-like regulation was revealed for some proteins. Quantitative or qualitative variation in a total of 386 proteins was influenced by genes assigned to at least one chromosomal region, while 66 % of all stained proteins were not found to be influenced by chromosome bins. Proteomics of deletion lines can, therefore, be used to simultaneously analyse the composition and genetics of a complex tissue, such as the wheat endosperm.

摘要

从小麦品种 Chinese Spring(CS)的胚乳中分离出白蛋白和球蛋白,以建立该标准小麦品种的蛋白质组参考图谱。通过二维凝胶电泳(2DE)检测到约 1145 个考马斯亮蓝染色斑点,其中 410 个斑点通过质谱分析和数据挖掘进行了鉴定。还通过 2DE 分离了 67 条 CS 缺失系的盐溶性胚乳蛋白(每条线四个凝胶)。将 268 张 2DE 凝胶的图像分析与 CS 参考蛋白质组进行比较,检测到由于染色体缺失导致胚乳蛋白的定性和定量变化。这些斑点的差异分析允许将结构或调节基因(编码 211 种蛋白质)定位在 21 条小麦染色体的片段上。此外,斑点体积定量变化的方差分析表明,391 种蛋白质的表达受一个或多个染色体 bin 控制,其中 262 种蛋白质的斑点体积显著增加,196 种蛋白质的斑点体积显著减少。一些蛋白质的斑点体积增加或减少是由许多染色体区域引起的,并且一些蛋白质显示出同源相似的调节。总共 386 种蛋白质的数量或质量变化受到至少一个染色体区域分配的基因的影响,而 66%的所有染色蛋白质未发现受到染色体 bin 的影响。因此,缺失系的蛋白质组学可以用于同时分析复杂组织(如小麦胚乳)的组成和遗传学。

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