Table_5_Genome-Wide Analysis of the C2 Domain Family in Soybean and Identification of a Putative Abiotic Stress Response Gene GmC2-148.xlsx

Plant C2 domain proteins play essential biological functions in numerous plants. In this study, 180 soybean C2 domain genes were identified by screening. Phylogenetic relationship analysis revealed that C2 domain genes fell into three distinct groups with diverged gene structure and conserved functional domain. Chromosomal location analysis indicated that C2 domain genes mapped to 20 chromosomes. The transcript profiles based on RNA-seq data showed that GmC2-58, GmC2-88, and GmC2-148 had higher levels of expression under salt, drought, and abscisic acid (ABA) treatments. GmC2-148, encoding a cell membrane-localized protein, had the highest level of response to various treatments according to real-time quantitative polymerase chain reaction (RT-qPCR) analysis. Under salt and drought stresses, the soybean plants with GmC2-148 transgenic hairy roots showed delayed leaf rolling, a higher content of proline (Pro), and lower contents of H2O2, O2– and malondialdehyde (MDA) compared to those of the empty vector (EV) plants. The results of transgenic Arabidopsis in salt and drought treatments were consistent with those in soybean treatments. In addition, the soybean plants with GmC2-148 transgenic hairy roots increased transcript levels of several abiotic stress-related marker genes, including COR47, NCDE3, NAC11, WRKY13, DREB2A, MYB84, bZIP44, and KIN1 which resulted in enhanced abiotic stress tolerance in soybean. These results indicate that C2 domain genes are involved in response to salt and drought stresses, and this study provides a genome-wide analysis of the C2 domain family in soybean.

植物C2结构域蛋白在众多植物中发挥着至关重要的生物学功能。本研究通过筛选鉴定了180个大豆C2结构域基因。系统发育关系分析显示，C2结构域基因可分为三个不同的组，这些组具有分化的基因结构和保守的功能域。染色体定位分析表明，C2结构域基因分布于20条染色体上。基于RNA-seq数据的转录分析揭示了GmC2-58、GmC2-88和GmC2-148在盐、干旱和脱落酸（ABA）处理下的表达水平较高。根据实时定量聚合酶链反应（RT-qPCR）分析，编码细胞膜定位蛋白的GmC2-148对各种处理具有最高的响应水平。在盐和干旱胁迫下，携带GmC2-148转基因毛根的大豆植株相较于空载体（EV）植株，表现出延迟的叶片卷曲、较高的脯氨酸（Pro）含量以及较低的H2O2、O2-和丙二醛（MDA）含量。在盐和干旱处理中，转基因拟南芥的结果与大豆处理结果一致。此外，携带GmC2-148转基因毛根的大豆植株提高了包括COR47、NCDE3、NAC11、WRKY13、DREB2A、MYB84、bZIP44和KIN1在内的多个与抗非生物胁迫相关标记基因的转录水平，从而增强了大豆的抗非生物胁迫能力。这些结果表明，C2结构域基因参与对盐和干旱胁迫的响应，本研究对大豆C2结构域家族进行了全基因组分析。