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One‐pot sample preparation approach for profiling spatial distribution of gibberellins in a single shoot of germinating cereal seeds
Author(s) -
Liu Cuimei,
Li Dongmei,
Li Jincheng,
Guo Zhenpeng,
Chen Yi
Publication year - 2019
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/tpj.14367
Subject(s) - gibberellin , shoot , derivatization , gas chromatography , chromatography , germination , sample preparation , gas chromatography–mass spectrometry , chemistry , mass spectrometry , agronomy , botany , biology
Summary Sample preparation remains a bottleneck in the rapid and reliable quantification of gibberellins ( GA s) for obtaining an insight into the physiological processes mediated by GA s. The challenges arise from not only the extremely low content of GA s in complex plant matrices, but the poor detectability of GA s 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 GA s 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 GA s 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 GA s. 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 GA s were detected and, among these, 11 GA s could be quantified in 0.50 mg fresh weight ( FW ) wheat shoot samples, and five GA s were quantified in only 0.15 mg FW developing seed samples of Arabidopsis thaliana . The distribution patterns of GA s 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 .