Fan Yapeng, Wang Ning, Wang Shuai, Yang Zhining, Chen Xiao, Lu Xuke, Huang Hui, Chen Xiugui, Zhao Lanjie, Zhang Menghao, Sun Yuping, Wang Junjuan, Guo Lixue, Wang Lidong, Song Ruize, Wang Jing, Zhang Xinrui, Yu Xin, Liu Yi, Zhou Xue-Rong, Meng Junting, Feng Keyun, Dai Maohua, Ye Wuwei
Institute of Cotton Research of Chinese Academy of Agricultural Sciences / Zhengzhou Research Base, State Key Laboratory of Cotton Bio-breeding and Integrated Utilization, School of Agricultural Sciences, Zhengzhou University / National Center of Technology Innovation for Comprehensive Utilization of Saline-Alkali Land, Anyang 455000, Henan, China.
Institute of Crop Sciences, Gansu Academy of Agricultural Sciences, Lanzhou 730070 Gansu, China.
J Adv Res. 2025 Jun 15. doi: 10.1016/j.jare.2025.06.018.
Ascorbic acid (AsA) is involved in plant responses to various abiotic stresses. However, its specific function in alkaline stress tolerance remains poorly understood. The L-galactono-1,4-lactone dehydrogenase (GLDH) gene is crucial for AsA synthesis, yet the precise role of GLDH in modulating plant resistance to alkaline stress has not been comprehensively characterized.
To investigate the role of GLDH genes in enhancing tolerance to alkaline stress.
Bioinformatics analysis of the GLDH gene family members was conducted, and an evolutionary tree was constructed using MEGA software. Cis-acting elements and gene structures were analyzed using TBtools. Gene expression levels were quantified by qRT-PCR, while the function of the GhGLDH35A gene was validated through VIGS (Virus-induced gene silencing) in cotton, heterologous overexpression in Arabidopsis thaliana, and complementation assays in yeast.
Our study investigated the effects of salt-alkaline stress on cotton and found that alkaline stress caused significantly more severe damage than salt stress. The GLDH family genes were identified and analyzed, revealing a high degree of evolutionary conservation. Most GhGLDH genes exhibited a positive response to alkaline stress and were regulated by light. Among them, GhGLDH35A, which is highly expressed within the GLDH family, was found to play a key role in conferring tolerance to alkaline stress. Subcellular localization analysis indicated that GhGLDH35A is localized in the mitochondria. Silencing of GhGLDH35A in cotton resulted in reduced tolerance to alkaline stress, disruption of ROS homeostasis, and impairment of photosynthesis and stomatal function. Conversely, overexpression of GhGLDH35A in Arabidopsis enhanced alkaline stress resistance by elevating AsA levels, increasing antioxidant enzyme activities to enhance ROS scavenging, sustaining photosynthesis, and promoting stomatal closure. Furthermore, heterologous expression of GhGLDH35A in yeast also improved its tolerance to alkaline stress.
GhGLDH35A positively regulates alkaline stress tolerance by enhancing antioxidant defenses and regulating stomatal movement.
抗坏血酸（AsA）参与植物对各种非生物胁迫的响应。然而，其在耐碱性胁迫中的具体功能仍知之甚少。L-半乳糖-1,4-内酯脱氢酶（GLDH）基因对AsA合成至关重要，但GLDH在调节植物对碱性胁迫抗性中的精确作用尚未得到全面表征。
研究GLDH基因在增强对碱性胁迫耐受性中的作用。
对GLDH基因家族成员进行生物信息学分析，使用MEGA软件构建进化树。使用TBtools分析顺式作用元件和基因结构。通过qRT-PCR定量基因表达水平，同时通过棉花中的病毒诱导基因沉默（VIGS）、拟南芥中的异源过表达以及酵母中的互补试验验证GhGLDH35A基因的功能。
我们的研究调查了盐碱胁迫对棉花的影响，发现碱性胁迫造成的损害比盐胁迫严重得多。对GLDH家族基因进行了鉴定和分析，发现其具有高度的进化保守性。大多数GhGLDH基因对碱性胁迫表现出阳性反应，并受光调控。其中，在GLDH家族中高表达的GhGLDH35A被发现赋予对碱性胁迫耐受性方面起关键作用。亚细胞定位分析表明GhGLDH35A定位于线粒体。棉花中GhGLDH35A的沉默导致对碱性胁迫的耐受性降低、活性氧稳态破坏以及光合作用和气孔功能受损。相反，拟南芥中GhGLDH35A的过表达通过提高AsA水平、增加抗氧化酶活性以增强活性氧清除、维持光合作用和促进气孔关闭来增强碱性胁迫抗性。此外，GhGLDH35A在酵母中的异源表达也提高了其对碱性胁迫的耐受性。
GhGLDH35A通过增强抗氧化防御和调节气孔运动来正向调节碱性胁迫耐受性。
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