School of Biological Science and Engineering, Hebei University of Economics and Business, Shijiazhuang 050061, China.
Biology Institute, Hebei Academy of Sciences, Shijiazhuang 050051, China.
Molecules. 2024 Feb 29;29(5):1074. doi: 10.3390/molecules29051074.
Cyclic adenosine monophosphate (cAMP) is an important second messenger in cells, mediating various stimulation signals such as the growth and development of organisms and stress and participating in regulating various biological processes of cells. This article explores the quantitative determination of cAMP in plants using High-Performance Liquid Chromatography (HPLC) and applies this method to analyzing the changes in cAMP content during the process of plant response to the bacterial quorum sensing signal -acyl homoserine lactone (AHL). Research has shown that the optimal detection conditions for HPLC are as follows: the chromatographic column is Venusil MP C18 (2), the mobile phase is methanol-water (0.1% trifluoroacetic acid) (:, 10:90), the detection wavelength is 259 nm, the column temperature is 35 °C, and the flow rate is 0.8 mL/min. The precision of the standard sample of this method is 98.21%, the precision of the sample is 98.87%, and the recovery rate is 101.067%. The optimal extraction conditions for cAMP in Arabidopsis are to use 15% methanol ultrasonic extraction for 10 min, followed by a 40 °C water bath for 4 h. Bacterial AHL signal processing can significantly stimulate an increase in cAMP levels in Arabidopsis leaves and roots. The establishment of HPLC detection methods for the cAMP content in plants is of great significance for in-depth research on the signal transduction mechanisms of plant-bacterial interactions.
环磷酸腺苷（cAMP）是细胞内一种重要的第二信使，介导着生物体的生长发育、应激等各种刺激信号，参与调节细胞的各种生物过程。本文采用高效液相色谱法（HPLC）定量测定植物中的 cAMP，并将该方法应用于分析植物对细菌群体感应信号——酰基高丝氨酸内酯（AHL）的响应过程中 cAMP 含量的变化。研究表明，HPLC 的最佳检测条件为：色谱柱 Venusil MP C18（2），流动相甲醇-水（0.1%三氟乙酸）（：，10:90），检测波长 259nm，柱温 35°C，流速 0.8mL/min。该方法标准样品的精密度为 98.21%，样品精密度为 98.87%，回收率为 101.067%。拟南芥中 cAMP 的最佳提取条件是采用 15%甲醇超声提取 10min，40°C水浴 4h。细菌 AHL 信号处理可以显著刺激拟南芥叶片和根系中 cAMP 水平的增加。建立植物中 cAMP 含量的 HPLC 检测方法，对于深入研究植物-细菌相互作用的信号转导机制具有重要意义。
