Quantification of near-attomole gibberellins in floral organs dissected from a single Arabidopsis thaliana flower
Quantification of near-attomole gibberellins in floral organs dissected from a single Arabidopsis thaliana flower
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从单个拟南芥花解剖的花器官中近阿托摩尔赤霉素的定量
DOI：
10.1111/tpj.13580
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发表时间：
2017-08-01
期刊：
影响因子：
7.2
通讯作者：
Chen, Yi
中科院分区：
文献类型：
--
作者：
Li, Dongmei;Guo, Zhenpeng;Chen, Yi
对鲜重低于毫克的极微小植物器官中的超痕量植物激素进行定量仍然存在方法学瓶颈。在内源性赤霉素 (GA) 的测定中，挑战变得更加严峻，内源性赤霉素 (GA) 是一类难以分离和检测的化合物。在此，开发了一种使用超高效液相色谱-串联质谱法并结合GA与乙醇中的N-(3-二甲基氨基丙基)-N-乙基碳二亚胺反应的衍生化技术的定量方法。该方法经验证能够测定花器官（约 80-250 克）（雌蕊、雄蕊、花瓣、萼片和花托）中的 GA，这些器官仅从拟南芥一朵花上解剖。在植物中，沿着非 13-羟基化途径和早期 13-羟基化途径，从花期 13、14 和 15 的花器官测量到 GA 的丰度模式存在显着差异。这使得我们能够从花的子层面深入了解 GA 如何影响花的发育。该方法表现出出色的检测限和定量限，分别低至 5.41 和 18.0 attomole，并提供了相当宽的线性范围（0.01 至 25 femtomole），线性系数高于 0.9961。该方法的精密度经过评估，日内测定的相对标准偏差低于 10.6%，日间测定的相对标准偏差低于 11.4%，回收率范围为 64.0% 至 107%。
There remains a methodological bottleneck in the quantification of ultra-trace plant hormones in very tiny plant organs at fresh weights below a milligram. The challenge becomes even more serious in the determination of endogenous gibberellins (GAs), which are a class of compounds that are difficult to separate and detect. Herein, a quantification method using ultra-high-performance liquid chromatography-tandem mass spectrometry was developed, combined with a derivatization technique in which GAs react with N-(3-dimethylaminopropyl)-N-ethylcarbodiimide in ethanol. The method was validated as capable of determining GAs in floral organs (about 80-250g) - pistil, stamens, petals, sepals and receptacle - which were dissected from only one flower of Arabidopsis thaliana. Substantially different abundance patterns of GAs were measured from the floral organs at floral stages 13, 14 and 15 along the non-13-hydroxylation pathway and the early 13-hydroxylation pathway in plants. This allows sub-flower-level insights into how GAs affect floral development. The method exhibited excellent limit of detection and limit of quantification down to 5.41 and 18.0 attomole, respectively, and offered a fairly wide linear range from 0.01 to 25 femtomole with linear coefficients above 0.9961. The precision of the method was evaluated with relative standard deviations below 10.6% for intra-day and 11.4% for inter-day assays, and recoveries ranged from 64.0% to 107%.