Department of Biosciences, Faculty of Science and Engineering, Teikyo University, Utsunomiya, 320-8551, Japan; Division of Integrated Science and Engineering, Graduate School of Science and Engineering, Teikyo University, Utsunomiya, 320-8551, Japan; Advanced Instrumental Analysis Center, Teikyo University, Utsunomiya, 320-8551, Japan.
Department of Biosciences, Faculty of Science and Engineering, Teikyo University, Utsunomiya, 320-8551, Japan; Division of Integrated Science and Engineering, Graduate School of Science and Engineering, Teikyo University, Utsunomiya, 320-8551, Japan.
Phytochemistry. 2021 Aug;188:112812. doi: 10.1016/j.phytochem.2021.112812. Epub 2021 May 18.
Jasmonic acid (JA) and its precursors are oxylipins derived from α-linolenic acid (αLA). Presumably, they are involved in the regulation of seed embryogenesis, dormancy, and germination. However, their spatial localization in the developing Phaseolus vulgaris L. (common bean) seeds has not been fully elucidated. Therefore, desorption electrospray ionization-mass spectrometry imaging (DESI-MSI) was performed to investigate their localization in the developing seeds. Peaks corresponding to the chemical formulae of αLA and 3-oxo-2-(2-(Z)-pentenyl)-cyclopentane-1-octanoic acid (OPC-8:0) were localized mainly in the radicle and seed coat, while that of 12-oxo-phytodienoic acid (OPDA) in the seed coat. This was consistent with the quantitative results obtained using liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) analysis. In contrast, DESI-tandem MSI (MS/MSI) and LC-ESI-MS/MS analyses showed that the effects of isomers on the DESI-MSI ion images were small for αLA and OPDA, but not for OPC-8:0. This indicated that DESI-MSI could accurately visualize αLA and OPDA, while DESI-MS/MSI was necessary to visualize OPC-8:0. The results demonstrated that free αLA and OPC-8:0 were abundant in the radicle and seed coat, while free OPDA was accumulated in the seed coat. Interestingly, the localization pattern of OPDA was similar to that of JA. In addition, compared to the concentrations of OPDA, the concentration of OPC-8:0 was lower in the seed coat and higher in the radicle. These results suggest that OPDA and/or JA play a biological role mainly in the seed coat, while OPC-8:0 is biologically active mainly in the radicle. Therefore, DESI-MSI coupled with LC-ESI-MS is a useful tool for spatial analysis of JA-related compounds in developing common bean seeds.
茉莉酸(JA)及其前体是由α-亚麻酸(αLA)衍生而来的氧化脂。它们可能参与调控种子胚胎发生、休眠和萌发。然而,它们在发育中的菜豆(普通豆)种子中的空间定位尚未完全阐明。因此,进行了解吸电喷雾电离-质谱成像(DESI-MSI)以研究它们在发育中的种子中的定位。对应于αLA 和 3-氧代-2-(2-(Z)-戊烯基)-环戊烷-1-辛酸(OPC-8:0)化学公式的峰主要定位于胚根和种皮中,而 12-氧代-植物二烯酸(OPDA)则定位于种皮中。这与使用液相色谱-电喷雾电离-质谱(LC-ESI-MS)分析获得的定量结果一致。相比之下,DESI-串联质谱成像(MS/MSI)和 LC-ESI-MS/MS 分析表明,异构体对 DESI-MSI 离子图像的影响对于 αLA 和 OPDA 较小,但对于 OPC-8:0 则较大。这表明 DESI-MSI 可以准确地可视化 αLA 和 OPDA,而 DESI-MS/MSI 则有必要可视化 OPC-8:0。结果表明,游离αLA 和 OPC-8:0 在胚根和种皮中含量丰富,而游离 OPDA则在种皮中积累。有趣的是,OPDA 的定位模式与 JA 相似。此外,与 OPDA 的浓度相比,OPC-8:0 在种皮中的浓度较低,在胚根中的浓度较高。这些结果表明,OPDA 和/或 JA 主要在种皮中发挥生物学作用,而 OPC-8:0 主要在胚根中发挥生物学作用。因此,DESI-MSI 与 LC-ESI-MS 相结合是一种用于分析发育中的普通豆种子中与 JA 相关化合物的空间分析的有用工具。