苹果液泡膜蔗糖转运蛋白基因MdSUT4的表达分析与功能鉴定

发布时间：2018-05-24 23:05

本文选题：苹果 + 液泡膜蔗糖转运蛋白　；参考：《园艺学报》2017年07期

【摘要】：以新疆红肉苹果杂种一代优系‘红脆1号’为试材,克隆MdSUT4并原核诱导获得其重组蛋白,对其进行生物信息学分析,测定其在不同组织及不同发育时期果实中的表达,通过亚细胞定位及转基因鉴定其功能。结果发现,MdSUT4编码的蛋白大小为55 kD左右,定位于8号染色体,由5个外显子和4个内含子组成;糖代谢相关基因系统进化树分析发现,MdSUT4与AtSUC4、PpSUT4在同一个进化支上;定量表达分析发现,MdSUT4在苹果的花、叶和幼果中均有较高的表达,且在果实发育中的表达水平与蔗糖含量显著负相关;MdSUT4启动子含有与糖信号及激素信号相关的顺式作用元件;在‘王林’苹果愈伤组织中过表达MdSUT4,能降低蔗糖含量,但却提高类黄酮含量;MdSUT4定位于‘王林’愈伤组织原生质体的液泡膜上。以上结果表明,MdSUT4可能参与了蔗糖从液泡膜中的外排,并可能促进类黄酮的合成。
[Abstract]:The recombinant protein of Xinjiang red apple hybrid 'Hongkui 1' was obtained by cloning MdSUT4 and inducing it into prokaryotic cells. The recombinant protein was analyzed by bioinformatics and its expression in different tissues and fruits at different developmental stages was determined. Its function was identified by subcellular localization and transgenic. The results showed that the protein encoding MdSUT4 was about 55kD, located on chromosome 8 and composed of 5 exons and 4 introns, and MdSUT4 and AtSUC4PpSUT4 were found to be on the same evolutionary branch by phylogenetic tree analysis. Quantitative expression analysis showed that MdSUT4 was highly expressed in apple flowers, leaves and young fruits, and the expression level of MdSUT4 in fruit development was negatively correlated with sucrose content. MdSUT4 promoter contained cis-acting elements related to sugar signal and hormone signal. Overexpression of MdSUT4 in 'Wanglin' apple callus decreased sucrose content, but increased flavonoid content. MdSUT4 was located on the vacuolar membrane of 'Wanglin' callus protoplast. These results suggest that MdSUT4 may be involved in the efflux of sucrose from the tonoplast and may promote the synthesis of flavonoids.
【作者单位】：山东农业大学园艺科学与工程学院/作物生物学国家重点实验室;
【分类号】：S661.1

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