PDF(Pubmed)
Abstract：
Legumin A is a seed storage protein that provides nutrients for seed germination. The purpose of this study was to describe the structure and expression pattern of the EuLEGA gene in Eucommia ulmoides Oliver (E. ulmoides) and to infer its functional role. The 1287 bp coding sequence of the EuLEGA CDS of the EuLEGA gene, encoding a protein containing 428 amino acid residues, was cloned. The structure predicted that the protein belonged to the RmlC (deoxythymidine diphosphates, dTDP)-4-dehydrorhamnose 3,5-epimerase)-like cupin conserved domain family, which contains both RmlC, a key enzyme for the synthesis of rhamnose and legumin A. The overexpression (OE) vector of the EuLEGA gene was constructed and genetically transformed into tobacco and E. ulmoides; the RNA interference (RNAi) vector of the EuLEGA gene was constructed and genetically transformed into E. ulmoides; and the contents of legumin A and rhamnose were detected. The results showed that the EuLEGA gene could significantly increase the content of legumin A in transgenic tobacco leaves and transgenic E. ulmoides regenerative buds, and the OE of this gene in E. ulmoides could promote an increase in rhamnose content. RNAi caused a significant decrease in the legumin A content in the regenerated buds of E. ulmoides. These was a significant increase in legumin A in the transgenic tobacco seeds, and these results indicate that the expression of the EuLEGA gene is closely related to the accumulation of legumin A. Subcellular localization studies revealed that EuLEGA is localized to the cytoplasm with the vacuolar membrane. Analysis of the EuLEGA gene expression data revealed that the expression level of the EuLEGA gene in the samaras was significantly greater than that in the leaves and stems. In addition, the study also demonstrated that GA3 can upregulate the expression levels of the EuLEGA gene, while ABA and MeJA can downregulate its expression levels.
摘要：
豆素A是一种种子贮藏蛋白，为种子萌发提供营养。本研究的目的是描述杜仲中EuLEGA基因的结构和表达模式（E.ulmoides)并推断其功能作用。EuLEGA基因的EuLEGACDS的1287bp编码序列，编码含有428个氨基酸残基的蛋白质，被克隆。该结构预测该蛋白质属于RmlC（脱氧胸苷二磷酸，dTDP）-4-脱水甘露糖3,5-差向异构酶）样cupin保守结构域家族，其中包含两个RmlC，构建了EuLEGA基因的过表达（OE）载体，并将其遗传转化到烟草和E.ulmoides中；构建了EuLEGA基因的RNA干扰（RNAi）载体，并将其遗传转化到E.结果表明,EuLEGA基因能显著提高转基因烟草叶片和转基因杜松子菜再生芽中豆腐素A的含量,并且该基因在E.ulmoides中的OE可以促进鼠李糖含量的增加。RNAi导致E.ulmoides再生芽中豆球蛋白A含量显着降低。这是转基因烟草种子中豆球蛋白A的显着增加，这些结果表明，EuLEGA基因的表达与豆类A的积累密切相关。亚细胞定位研究表明，EuLEGA定位于液泡膜的细胞质中。对EuLEGA基因表达数据的分析显示，EuLEGA基因在萨马拉中的表达水平显着高于在叶和茎中的表达水平。此外,该研究还表明GA3可以上调EuLEGA基因的表达水平，而ABA和MeJA可以下调其表达水平。
