Functional characterization of the A. thaliana Aldehyde Dehydrogenase gene family

许多醛在低浓度下对生物体有毒。因此，严格控制代谢醛水平是很重要的。醛脱氢酶（ALDHs）形成一个大的蛋白质家族，并催化各种醛底物的不可逆氧化。植物具有多种不同的ALDHs，但其生物学作用的信息有限。拟开展的研究项目旨在确定拟南芥ALDH基因家族的生物学功能，重点关注3类ALDH、BADH和瘤体反应性ALDH基因的成员，因为它们在渗透胁迫适应中具有潜在作用。将采用以下实验方法：分离鉴定出的ALDH基因的敲除突变，研究其对生理和形态特征的影响。此外，还将构建具有修饰ALDH基因表达的转基因植株。在确定的生理条件下，分析野生型、突变型和转基因植物中ALDH的表达。重组蛋白将用于确定aldh的酶学性质。为了鉴定假定的体内底物，野生型、突变型和转基因植物的代谢物谱分析和底物捕获策略将在未来的方法中使用。

Many aldehydes are toxic for organisms at low concentrations. It is therefore important that metabolic aldehyde levels are strictly controlled. Aldehyde dehydrogenases (ALDHs) form a large protein family and catalyze the irreversible oxidation of a wide range of aldehyde substrates. Plants possess a variety of distinct ALDHs, but information on their biological role is limited. The aim of the proposed research project is to determine the biological function of the ALDH gene family from A. thaliana focusing on members of the class-3 ALDH, BADH and turgor-responsive ALDH genes, because of their potential role in osmotic stress adaptation. The following experimental approaches will be pursued: Knock-Out mutations for the identified ALDH genes will be isolated to study their effects on physiological and morphological characteristics. In addition, transgenic plants with a modified ALDH gene expression will be constructed. ALDH expression will be analyzed in wild-type, mutant and transgenic plants under defined physiological conditions. Recombinant proteins will be used to determine enzymatic properties of the ALDHs. To identify putative in vivo substrates, metabolite profiling of wild-type, mutant and transgenic plants and a substrate trapping strategy will be used in future approaches.