College of Life Science, Capital Normal University, Beijing 100048, China.
BMC Plant Biol. 2012 Aug 19;12:147. doi: 10.1186/1471-2229-12-147.
The analyses of protein synthesis, accumulation and regulation during grain development in wheat are more complex because of its larger genome size compared to model plants such as Arabidopsis and rice. In this study, grains from two wheat cultivars Jimai 20 and Zhoumai 16 with different gluten quality properties were harvested at five development stages, and were used to displayed variable expression patterns of grain proteins.
Proteome characterization during grain development in Chinese bread wheat cultivars Jimai 20 and Zhoumai 16 with different quality properties was investigated by 2-DE and tandem MALDI-TOF/TOF-MS. Identification of 117 differentially accumulated protein spots representing 82 unique proteins and five main expression patterns enabled a chronological description of wheat grain formation. Significant proteome expression differences between the two cultivars were found; these included 14 protein spots that accumulated in both cultivars but with different patterns and 27 cultivar-different spots. Among the cultivar-different protein spots, 14 accumulated in higher abundance in Jimai 20 than in Zhoumai 16, and included NAD-dependent isocitrate dehydrogenase, triticin precursor, LMW-s glutenin subunit and replication factor C-like protein. These proteins are likely to be associated with superior gluten quality. In addition, some proteins such as class II chitinase and peroxidase 1 with isoforms in developing grains were shown to be phosphorylated by Pro-Q Diamond staining and phosphorprotein site prediction. Phosphorylation could have important roles in wheat grain development. qRT-PCR analysis demonstrated that transcriptional and translational expression patterns of many genes were significantly different.
Wheat grain proteins displayed variable expression patterns at different developmental stages and a considerable number of protein spots showed differential accumulation between two cultivars. Differences in seed storage proteins were considered to be related to different quality performance of the flour from these wheat cultivars. Some proteins with isoforms were phosphorylated, and this may reflect their importance in grain development. Our results provide new insights into proteome characterization during grain development in different wheat genotypes.
与拟南芥和水稻等模式植物相比，小麦的基因组更大，因此在分析其发育过程中的蛋白质合成、积累和调控时更为复杂。本研究以面筋品质特性不同的两个小麦品种济麦 20 和周麦 16 的籽粒为材料，在五个发育阶段进行收获，用于展示籽粒蛋白质的可变表达模式。
利用 2-DE 和串联 MALDI-TOF/TOF-MS 对具有不同品质特性的中国面包小麦品种济麦 20 和周麦 16 的籽粒发育过程中的蛋白质组进行了分析。鉴定了 117 个差异表达的蛋白点，代表 82 种独特的蛋白质和 5 种主要的表达模式，从而可以对小麦籽粒形成进行时间描述。在两个品种之间发现了显著的蛋白质组表达差异；其中包括在两个品种中都积累但模式不同的 14 个蛋白点和 27 个品种特异的蛋白点。在品种特异的蛋白点中，有 14 个在济麦 20 中的积累量高于周麦 16，包括 NAD 依赖的异柠檬酸脱氢酶、麦谷蛋白前体、LMW-s 谷蛋白亚基和复制因子 C 样蛋白。这些蛋白质可能与较好的面筋品质有关。此外，一些蛋白质，如发育籽粒中的 II 类几丁质酶和过氧化物酶 1，通过 Pro-Q Diamond 染色和磷酸化蛋白位点预测显示出磷酸化。磷酸化可能在小麦籽粒发育中具有重要作用。qRT-PCR 分析表明，许多基因的转录和翻译表达模式差异显著。
小麦籽粒蛋白质在不同发育阶段表现出不同的表达模式，许多蛋白点在两个品种之间表现出差异积累。种子贮藏蛋白的差异被认为与这些小麦品种面粉的不同品质性能有关。一些具有同工型的蛋白质发生磷酸化，这可能反映了它们在籽粒发育中的重要性。本研究结果为不同小麦基因型籽粒发育过程中的蛋白质组学研究提供了新的见解。
