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Cloning and functional characterization of the legumin A gene (EuLEGA) from Eucommia ulmoides Oliver

文献类型： 外文期刊

作者： Zheng, Lina 1 ; Zhao, De-Gang 1 ;

作者机构： 1.Guizhou Univ, Coll Life Sci Inst Agro Bioengn, Key Lab Plant Resource Conservat & Germplasm Inno, Minist Educ, Guiyang 550025, Guizhou, Peoples R China

2.Guizhou Acad Agr Sci, Biotechnol Inst Guizhou, Guizhou Plant Conservat Technol Ctr, Guiyang 550006, Guizhou, Peoples R China

关键词： Eucommia ulmoides; Legumin A; Legumin A-encoding gene; Transgene

期刊名称：SCIENTIFIC REPORTS （ 影响因子：3.8； 五年影响因子：4.3 ）

ISSN： 2045-2322

年卷期： 2024 年 14 卷 1 期

页码：

收录情况： SCI

摘要： 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 GA(3) can upregulate the expression levels of the EuLEGA gene, while ABA and MeJA can downregulate its expression levels.

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