Guo Weileng, Sun Yuanqing, Chai Juqi, Liu Lei, Li Jiaqi, Guo Rui, Guo Changhong
Key Laboratory of Molecular and Cytogenetics, College of Life Science and Technology, Harbin Normal University, Harbin 150025, China.
Key Laboratory of Molecular and Cytogenetics, College of Life Science and Technology, Harbin Normal University, Harbin 150025, China.
Gene. 2025 Sep 5;963:149609. doi: 10.1016/j.gene.2025.149609. Epub 2025 Jun 2.
Trehalose-6-phosphate synthase (TPS) is an indispensable enzyme in the trehalose synthesis pathway, which plays an active role in plant responses to abiotic stress. However, the identification of the TPS gene family in alfalfa and its effects under saline-alkali stress remain unclear. This study identified 38 MsTPS genes that could be categorized into two classes. Class I genes had more introns than did Class II genes. Class II genes possessed the full motif, whereas Class I genes lacked motif 8. The evolutionary pattern of MsTPS genes primarily manifested as fragment duplication events. The promoter regions of the MsTPS genes were enriched with multiple hormone- and stress-response cis-acting regulatory elements. Transcriptome analyses revealed that nine MsTPS genes were upregulated under saline-alkali stress. A qRT-PCR analysis revealed that MsTPS16 exhibited the most significant upregulated expression. Subcellular localization analyses revealed that the MsTPS16 protein was mainly localized in the cytoplasm, vacuolar membrane, and cell membrane. The heterologous expression of MsTPS16 promoted yeast growth under saline-alkali stress, and its overexpression in alfalfa hairy roots reduced the reactive oxygen species content and enhanced the expression of genes encoding the key enzymes of the antioxidant pathway under saline-alkali stress. This study provides comprehensive insights into the functional characterization of the MsTPS gene family and highlights a potentially influential gene involved in tolerance to saline-alkali stress in alfalfa.
海藻糖-6-磷酸合酶（TPS）是海藻糖合成途径中不可或缺的一种酶，在植物对非生物胁迫的响应中发挥着积极作用。然而，苜蓿中TPS基因家族的鉴定及其在盐碱胁迫下的作用仍不清楚。本研究鉴定出38个MsTPS基因，可分为两类。I类基因比II类基因具有更多的内含子。II类基因具有完整的基序，而I类基因缺乏基序8。MsTPS基因的进化模式主要表现为片段重复事件。MsTPS基因的启动子区域富含多种激素和胁迫响应顺式作用调控元件。转录组分析表明，9个MsTPS基因在盐碱胁迫下上调表达。qRT-PCR分析表明，MsTPS16表现出最显著的上调表达。亚细胞定位分析表明，MsTPS16蛋白主要定位于细胞质、液泡膜和细胞膜。MsTPS16的异源表达促进了盐碱胁迫下酵母的生长，其在苜蓿毛状根中的过表达降低了活性氧含量，并增强了盐碱胁迫下抗氧化途径关键酶编码基因的表达。本研究为MsTPS基因家族的功能特性提供了全面的见解，并突出了一个可能影响苜蓿耐盐碱胁迫的基因。
