Department Cell Biology, Plant Physiology, University of Salzburg, Hellbrunnerstr. 34, Salzburg, 5020, Austria.
Plant J. 2013 Apr;74(2):239-47. doi: 10.1111/tpj.12116. Epub 2013 Feb 13.
Numerous nucleotide sugars are needed in plants to synthesize cell wall polymers and glycoproteins. The de novo synthesis of nucleotide sugars is of major importance. During growth, however, some polymers are broken down to monosaccharides. Reactivation of these sugars into nucleotide sugars occurs in two steps: first, by a substrate-specific sugar-1-kinase and, second, by UDP-sugar-pyrophosphorylase (USP), which has broad substrate specificity. A knock-out of the USP gene results in non-fertile pollen. By using various genetic complementation approaches we obtained a strong (>95%) knock-down line in USP that allowed us to investigate the physiological role of the enzyme during the life cycle. Mutant plants show an arabinose reduction in the cell wall, and accumulate mainly two sugars, arabinose and xylose, in the cytoplasm. The arabinogalactanproteins in usp mutants show no significant reduction in size. USP is also part of the myo-inositol oxygenation pathway to UDP-glucuronic acid; however, free glucuronic acid does not accumulate in cells, suggesting alternative conversion pathways of this monosaccharide. The knock-down plants are mostly sterile because of the improper formation of anthers and pollen sacks.
植物中需要大量的核苷酸糖来合成细胞壁聚合物和糖蛋白。核苷酸糖的从头合成非常重要。然而,在生长过程中,一些聚合物会分解成单糖。这些糖重新激活为核苷酸糖需要经过两步反应:首先,通过特异性糖激酶;然后,通过 UDP-糖焦磷酸化酶(USP),该酶具有广泛的底物特异性。USP 基因的敲除会导致花粉不育。通过使用各种遗传互补方法,我们获得了 USP 的强敲低 (>95%)系,使我们能够在生命周期内研究该酶的生理作用。突变体植物的细胞壁阿拉伯糖减少,细胞质中主要积累两种糖,阿拉伯糖和木糖。usp 突变体中的阿拉伯半乳聚糖蛋白大小没有明显减少。USP 也是肌醇氧化途径向 UDP-葡萄糖醛酸的一部分;然而,细胞中没有游离葡萄糖醛酸积累,这表明该单糖可能存在替代转化途径。由于花药和花粉囊的形成不当,敲低植物大多不育。