Li Yiran, Lian Boying, Gao Yanna, Zhang Nan, Ahmad Adeel, Wang Mengyan, Wu Hongmei, Wu Jiajia, Yang Fan, Wang Baoquan, Hao Fushun, Wei Hengling
State Key Laboratory of Cotton Bio-breeding and Integrated Utilization, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang, Henan 455000, China; National Key Laboratory of Cotton Bio-breeding and Integrated Utilization, School of Life Sciences, Henan University, Kaifeng 475004, China.
State Key Laboratory of Cotton Bio-breeding and Integrated Utilization, Institute of Cotton Research, Chinese Academy of Agricultural Sciences, Anyang, Henan 455000, China.
Plant Sci. 2025 Oct;359:112601. doi: 10.1016/j.plantsci.2025.112601. Epub 2025 Jun 7.
Aquaporins (AQPs) facilitate transmembrane water transport and play a crucial role in plant adaptation to diverse abiotic stress conditions. Plasma membrane Intrinsic Proteins (PIPs) are a crucial subgroup of aquaporins that facilitate water transport across plant cell membrane. However, the role of PIPs under salt stress responses in cotton remains partially unexplored. In this study, we found that the expression level of the aquaporin gene GhPIP2;1 was significantly upregulated under salt stress at three-leaf stage in upland cotton. The overexpression of GhPIP2;1 in Arabidopsis thaliana and virus-induced gene silencing (VIGS) in upland cotton evidenced that GhPIP2;1 positively regulate salt tolerance function. Biochemical analysis including superoxide dismutase (SOD), peroxidase (POD) activities and malondialdehyde (MDA) content measurements indicated that GhPIP2;1 could enhance stress tolerance by regulating the reactive oxygen species (ROS). The Electrophoretic mobility shift assay (EMSA) and dual luciferase reporter assay confirmed that GhNAC072 specifically binds to the promoter region of GhPIP2;1 and enhance the expression of GhPIP2;1. Phenotypic observations and physiological measurements of GhNAC072 overexpressed Arabidopsis and silenced cotton plants demonstrated that GhNAC072 positively responds to salt stress. The expression level of GhPIP2;1 was significantly decreased in GhNAC072-silenced plants, indicating that GhNAC072 enhances salt tolerance by regulating GhPIP2;1 expression. In summary, this study systematically confirmed that GhPIP2;1 may involve in salt tolerance mechanism in cotton and its expression is regulated by GhNAC072. These results will provide gene resources for salt tolerance and enrich the knowledge on the mechanism of salt tolerance in cotton.
水通道蛋白（AQPs）促进跨膜水分运输，在植物适应多种非生物胁迫条件中发挥关键作用。质膜内在蛋白（PIPs）是水通道蛋白的一个关键亚组，有助于水分跨植物细胞膜运输。然而，PIPs在棉花盐胁迫响应中的作用仍部分未被探索。在本研究中，我们发现陆地棉三叶期盐胁迫下，水通道蛋白基因GhPIP2;1的表达水平显著上调。GhPIP2;1在拟南芥中的过表达以及陆地棉中的病毒诱导基因沉默（VIGS）证明，GhPIP2;1正向调节耐盐功能。包括超氧化物歧化酶（SOD）、过氧化物酶（POD）活性和丙二醛（MDA）含量测定在内的生化分析表明，GhPIP2;1可通过调节活性氧（ROS）增强胁迫耐受性。电泳迁移率变动分析（EMSA）和双荧光素酶报告基因分析证实，GhNAC072特异性结合GhPIP2;1的启动子区域并增强GhPIP2;1的表达。对过表达GhNAC072的拟南芥和沉默GhNAC072的棉花植株的表型观察和生理测量表明，GhNAC072对盐胁迫有正向响应。GhNAC072沉默植株中GhPIP2;1的表达水平显著降低，表明GhNAC072通过调节GhPIP2;1的表达增强耐盐性。总之，本研究系统地证实了GhPIP2;1可能参与棉花的耐盐机制，其表达受GhNAC072调控。这些结果将为棉花耐盐提供基因资源，并丰富棉花耐盐机制的知识。
