Yang Xiaopeng, Luo Fei, Wang Haiyang, Liu Yu, Zhang Shiyi, Yan Dingwei, Wei Yuewei, Li Junying, Zhang Di, Ji Xiaoming
College of Tobacco Science, Henan Agricultural University, Zhengzhou, 450046, China.
College of Tobacco Science, Henan Agricultural University, Zhengzhou, 450046, China.
Anal Chim Acta. 2025 Jan 22;1336:343478. doi: 10.1016/j.aca.2024.343478. Epub 2024 Nov 28.
Nitroxyl (HNO) is an emerging signaling molecule that plays a significant regulatory role in various aspects of plant biology, including stress responses and developmental processes. However, understanding the precise actions of HNO in plants has been challenging due to the absence of highly sensitive and real-time in situ monitoring tools. Consequently, it is crucial to develop effective and accurate detection methods for HNO. Establishing such methodologies will enable researchers to elucidate the functional roles of HNO in plant physiological processes, thereby advancing our knowledge of plant resilience and adaptation under environmental stressors.
Herein, we successfully constructed a near-infrared fluorescent probe, DCIF-HNO, based on the dicyanoisophorone platform as fluorophore and 2-(diphenylphosphino)benzoate as HNO recognition site for identifying HNO in plants. Probe DCIF-HNO exhibited rapid response, excellent selectivity, and high sensitivity to HNO in vitro spectroscopic tests, while also demonstrating low toxicity and biocompatibility. A rapid and portable smartphone sensing platform for HNO in actual samples was successfully constructed based on probe DCIF-HNO and color recognition application. Moreover, probe DCIF-HNO was successfully applied to plant cells and tissues, enabling real-time visualization and detection of HNO and revealing the complex network of HNO interactions during HS/NO crosstalk in plants. Furthermore, the increase in HNO levels in plants response to high salt and Cr stress was observed using probe DCIF-HNO. Transcriptome sequencing and differential metabolites analysis were employed to gain insight into the mechanism of HNO production under Cr stress.
Due to the optical properties and high-resolution imaging capabilities of DCIF-HNO, this study offers a novel framework for elucidating the signaling role of HNO in plant stress responses. The precise visualization of HNO dynamics enhances our understanding of the complex molecular pathways involved in plant adaptation to abiotic stressors. This research not only advances plant physiology but also has significant implications for developing strategies to enhance agricultural resilience in challenging environmental conditions.
硝酰基(HNO)是一种新兴的信号分子,在植物生物学的各个方面发挥着重要的调节作用,包括应激反应和发育过程。然而,由于缺乏高灵敏度和实时原位监测工具,了解HNO在植物中的精确作用一直具有挑战性。因此,开发有效的HNO检测方法至关重要。建立此类方法将使研究人员能够阐明HNO在植物生理过程中的功能作用,从而增进我们对植物在环境应激源下的恢复力和适应性的认识。
在此,我们成功构建了一种基于二氰基异佛尔酮平台作为荧光团、2-(二苯基膦基)苯甲酸酯作为HNO识别位点的近红外荧光探针DCIF-HNO,用于识别植物中的HNO。探针DCIF-HNO在体外光谱测试中对HNO表现出快速响应、优异的选择性和高灵敏度,同时还具有低毒性和生物相容性。基于探针DCIF-HNO和颜色识别应用,成功构建了一个用于实际样品中HNO的快速便携式智能手机传感平台。此外,探针DCIF-HNO成功应用于植物细胞和组织,能够实时可视化和检测HNO,并揭示植物中HS/NO串扰过程中HNO相互作用的复杂网络。此外,使用探针DCIF-HNO观察到植物对高盐和铬胁迫的反应中HNO水平的增加。采用转录组测序和差异代谢物分析来深入了解铬胁迫下HNO产生的机制。
由于DCIF-HNO的光学性质和高分辨率成像能力,本研究为阐明HNO在植物应激反应中的信号作用提供了一个新框架。HNO动态的精确可视化增强了我们对植物适应非生物应激源所涉及的复杂分子途径的理解。这项研究不仅推动了植物生理学的发展,而且对制定在具有挑战性的环境条件下增强农业恢复力的策略也具有重要意义。