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One-pot sample preparation approach for profiling spatial distribution of gibberellins in a single shoot of germinating cereal seeds

文献类型： 外文期刊

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

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

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

3.North China Univ Sci & Technol, Sch Pharm, Tangshan 063210, Hebei, Peoples R China

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

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

关键词： gibberellins; one-pot sample preparation approach; N-(3-dimethylaminopropyl)-N '-ethylcarbodiimide; spatial distribution; single shoot; UHPLC-ESI-MS; MS; Triticum aestivum; Oryza sativa; Zea mays; technical advance

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

ISSN： 0960-7412

年卷期： 2019 年 99 卷 5 期

页码：

收录情况： SCI

摘要： Sample preparation remains a bottleneck in the rapid and reliable quantification of gibberellins (GAs) for obtaining an insight into the physiological processes mediated by GAs. The challenges arise from not only the extremely low content of GAs in complex plant matrices, but the poor detectability of GAs by mass spectrometry (MS) in negative ion mode. In an effort to solve these urgent difficulties, we present a spatial-resolved analysis method to investigate the distribution of GAs in tiny plant tissues based on a simplified one-pot sample preparation approach coupled with ultrahigh-performance liquid chromatography-tandem MS. By integrating extraction and derivatization into one step, target GAs were effectively extracted from plant materials and simultaneously reacted with N-(3-dimethylaminopropyl)-N '-ethylcarbodiimide, the sample preparation time was largely shortened, the probability of sample loss was minimized and the detection sensitivity of MS was also greatly improved compared with underivatized GAs. Under optimal conditions, the method was validated from the quantification linearity, limits of detection and limits of quantification in the presence of plant matrices, recoveries, and precision. With the proposed method, 15 endogenous GAs were detected and, among these, 11 GAs could be quantified in 0.50 mg fresh weight (FW) wheat shoot samples, and five GAs were quantified in only 0.15 mg FW developing seed samples of Arabidopsis thaliana. The distribution patterns of GAs along both the non-13-hydroxylation pathway and the early 13-hydroxylation pathway in a single shoot of germinating wheat, rice and maize seeds were finally profiled with a spatial resolution down to approximately 1 mm(2).

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