Table_6_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，鉴定出180个大豆C2结构域基因。系统发育关系分析表明，C2结构域基因可分为三个不同的组别，这些组别在基因结构上存在分化，而在功能域上则保持保守。染色体定位分析显示，C2结构域基因分布于20条染色体上。基于RNA测序数据的转录谱分析显示，在盐、干旱和脱落酸（ABA）处理下，GmC2-58、GmC2-88和GmC2-148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在多种处理条件下表现出最高的反应水平。在盐和干旱胁迫下，与空载体（EV）植物相比，携带GmC2-148转基因毛根的大豆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植株的处理结果一致。此外，携带GmC2-148转基因毛根的大豆植株中，与多种非生物胁迫相关标记基因（如COR47、NCDE3、NAC11、WRKY13、DREB2A、MYB84、bZIP44和KIN1）的转录水平增加，从而增强了大豆的非生物胁迫耐受性。这些结果揭示了C2结构域基因在应对盐和干旱胁迫中的作用，并为大豆C2结构域家族的全基因组分析提供了依据。