植物根际促生菌发酵液中植物激素含量的液液萃取-高效液相色谱测定法  

Determination of plant hormones in bacterial fermentation products of plant growth-promoting rhizobacteria by liquid-liquid extraction high-performance liquid chromatography

作　　者：刘晓婷 姚拓[1] 马亚春 柴加丽 LIU Xiaoting;YAO Tuo;MA Yachun;CHAI Jiali(College of Grassland Science,Gansu Agricultural University,Lanzhou 730070,Gansu,China)

机构地区：[1]甘肃农业大学草业学院,甘肃兰州730070

出　　处：《草业科学》2022年第10期2222-2228,共7页Pratacultural Science

基　　金：高寒草地退化植被恢复与土壤改良菌剂研发与示范(GSAU-TSYF-2021-011);甘肃省教育厅“双一流”科研重点项目(GSSYLXM-02)。

摘　　要：植物根际促生菌分泌的植物激素在植物生长过程中发挥着重要作用,能够调节植物生长、发育及抗逆的各个过程。了解植物根际促生菌分泌的植物激素含量,对优良促生菌资源的筛选至关重要。本研究建立了高效液相色谱法以快速准确测定植物根际促生菌发酵液中植物激素含量。样品用超纯水-乙酸乙酯(体积比1:2)提取,以Agilent ZORBAX Eclipse Plus C色谱柱分离,采用体积比45:55的甲醇-水(含0.2%冰乙酸)作为流动相,流速为0.8mL·min^(-1),柱温为30℃,进样体积为10μL,在254 nm检测波长下进行检测。结果表明:3种植物激素在测定的浓度范围内线性关系良好,相关系数(R~2)均大于0.997,样品的平均加标回收率为88.02%~108.40%,相对标准偏差(n=3)为0.02%~6.51%。该方法简单、快速,适用于植物根际促生菌中3种植物激素同时测定,研究结果为今后微生物肥料的研制应用提供了菌种资源与参考。Plant hormones secreted by plant growth-promoting rhizobacteria play an important role in plant growth, as they regulate the various processes related to plant growth, development, and stress resistance. Understanding the content of plant hormones in bacterial fermentation products of plant growth-promoting rhizobacteria is crucial for the screening of excellent growth-promoting bacterial resources. Therefore, this study aims to develop a simple high-performance liquid chromatographic method for the simultaneous detection of gibberellin A3, indole-3-acetic acid, and trans-Zeatin. The samples were extracted with ultrapure water and ethyl acetate(1 : 2, by volume) and then separated by Agilent ZORBAX Eclipse Plus C18 chromatographic column. The mobile phase was 45% methanol and 55% ultrapure water(containing 0.2% glacial acetic acid) and the detection wavelength was set at 254 nm. The flow rate was 0.8 mL·min^(-1), the column temperature of 30 ℃ and the injection volume of 10 μL were adopted. Results indicated that the linear relationship of the three plant hormones is good within their linear range and the correlation coefficients were not less than 0.997. The average recoveries of this method ranged from 88.02% to 108.40% with relative standard deviation of 0.02%~ 6.51%. This method is simple, rapid, and suitable for simultaneous determination of several plant hormones in plant growth-promoting rhizobacteria, which could provide a good reference and bacterial strain resources for the development and application of microbial fertilizer in the future.

关 键 词：植物根际促生菌 高效液相色谱法 植物激素 定量测定 赤霉素 吲哚乙酸 玉米素 

分 类 号：O657.72[理学—分析化学] TQ446[理学—化学] Q946.885[化学工程—化学肥料工业]