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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。
The recombinant protein of MdSUT4 was cloned and prokaryotic induced by Xinjiang red meat apple hybrid genotype “Huancui 1”. The recombinant protein was analyzed by bioinformatics and the expression of MdSUT4 in different tissues and fruits at different developmental stages , Through subcellular localization and transgene identification of its function. The results showed that the size of MdSUT4 encoded protein was about 55 kD and located on chromosome 8, which consisted of 5 exons and 4 introns. Phylogenetic tree analysis showed that MdSUT4 was closely related to AtSUC4 and PpSUT4 A branch of evolution. Quantitative expression analysis showed that MdSUT4 was highly expressed in the flowers, leaves and young fruits of apple, and its expression level in fruit development was negatively correlated with sucrose content. And hormone signal related cis-acting elements. Overexpression of MdSUT4 in ’Wanglin’ apple callus could reduce sucrose content but increase flavonoid content. MdSUT4 was localized in the vacuole of ’Wanglin’ callus protoplast Membrane. The above results indicate that MdSUT4 may be involved in the efflux of sucrose from the vacuolar membrane and may promote the synthesis of flavonoids.