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Quantification of near-attomole gibberellins in floral organs dissected from a single Arabidopsis thaliana flower

文献类型： 外文期刊

作者： Li, Dongmei 1 ; Guo, Zhenpeng 1 ; Liu, Cuimei 1 ; Li, Jincheng 3 ; Xu, Wenzhong 4 ; Chen, Yi 1 ;

作者机构： 1.Chinese Acad Sci, Inst Chem, CAS Res Educ Ctr Excellence Mol Sci, Key Lab Analyt Chem Living Biosyst, Beijing 100190, Peoples R China

2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China

3.Chinese Acad Fishery Sci, Beijing 100141, Peoples R China

4.Chinese Acad Sci, Inst Bot, Key Lab Plant Resources, Beijing 100093, Peoples R China

5.Beijing Natl Lab Mol Sci, Beijing 100190, Peoples R China

关键词： gibberellins;UHPLC-ESI-MS;MS;tiny fresh plant organ;quantification;Arabidopsis thaliana flower

期刊名称：PLANT JOURNAL （ 影响因子：6.417； 五年影响因子：7.627 ）

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收录情况： SCI

摘要： 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%.

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