Serrato Antonio J, Cejudo Francisco J
Instituto de Bioquímica Vegetal y Fotosíntesis, Centro de Investigaciones Científicas Isla de la Cartuja, Avda Américo Vespucio s/n, 41092 Sevilla, Spain.
Planta. 2003 Jul;217(3):392-9. doi: 10.1007/s00425-003-1009-4. Epub 2003 Mar 15.
Thioredoxin h (Trx h) proteins are ubiquitous in all wheat organs, but show the highest accumulation in mature seeds. This distribution suggests the expression of Trx h during seed development. In the present study, we have analyzed the pattern of Trx h expression in developing wheat ( Triticum aestivum L.) seeds. Northern blot analysis detected a single band at any stage of development, which corresponded to the expression of at least two genes, TrxhA and TrxhB, as shown by competitive reverse transcription-polymerase chain reaction experiments. The analysis of the content of Trx h polypeptides showed the highest content in the embryo. The spatial pattern of accumulation of these proteins was established by immunocytological techniques. At early stages of development Trx h proteins localized to maternal tissues (nucellus projection cells and pedicel), the route of transport of nutrients to the developing endosperm. In the endosperm, Trx h proteins accumulated at a high level in the aleurone layer. At later stages of development, during seed maturation, Trx h proteins localized predominantly to the nucleus of both aleurone and scutellum cells, a feature exclusive of these seed tissues. The nuclear localization of Trx h proteins was associated with oxidative stress in these tissues, as shown by in situ staining of superoxide radicals in developing and germinating seeds.
硫氧还蛋白h(Trx h)在小麦所有器官中普遍存在,但在成熟种子中积累量最高。这种分布情况表明Trx h在种子发育过程中表达。在本研究中,我们分析了发育中的小麦(普通小麦)种子中Trx h的表达模式。Northern杂交分析在发育的任何阶段都检测到一条带,竞争性逆转录-聚合酶链反应实验表明,该带对应于至少两个基因TrxhA和TrxhB的表达。Trx h多肽含量分析显示,胚中的含量最高。通过免疫细胞技术确定了这些蛋白质积累的空间模式。在发育早期,Trx h蛋白定位于母体组织(珠心突起细胞和花梗),这是营养物质向发育中的胚乳运输的途径。在胚乳中,Trx h蛋白在糊粉层中大量积累。在发育后期,即种子成熟期间,Trx h蛋白主要定位于糊粉层和盾片细胞的细胞核,这是这些种子组织特有的特征。发育中和萌发种子中超氧自由基的原位染色表明,Trx h蛋白的核定位与这些组织中的氧化应激有关。