Balcke Gerd U, Bennewitz Stefan, Zabel Sebastian, Tissier Alain
Department of Cell and Metabolic Biology, Institute of Plant Biochemistry, Weinberg 3, Halle, 06120, Germany.
Methods Mol Biol. 2014;1153:189-202. doi: 10.1007/978-1-4939-0606-2_13.
Plant glandular trichomes are specialized secretory structures located on the surface of the aerial parts of plants with large biosynthetic capacity, often with terpenoids as output molecules. The collection of plant trichomes requires a method to separate trichomes from leaf epidermal tissues. For metabolite profiling, trichome tissue needs to be rapidly quenched in order to maintain the indigenous state of intracellular intermediates. Appropriate extraction and chromatographic separation methods must be available, which address the wide-ranging polarity of metabolites. In this chapter, a protocol for trichome harvest using a frozen paint brush is presented. A work flow for broad-range metabolite profiling using LC-MS(2) analysis is described, which is applicable to assess very hydrophilic isoprenoid precursors as well as more hydrophobic metabolites from trichomes and other plant tissues.
植物腺毛是位于植物地上部分表面的特殊分泌结构,具有强大的生物合成能力,其输出分子通常为萜类化合物。收集植物腺毛需要一种从叶片表皮组织中分离腺毛的方法。对于代谢物谱分析,腺毛组织需要迅速淬灭,以维持细胞内中间体的原始状态。必须有合适的提取和色谱分离方法,以应对代谢物广泛的极性。在本章中,介绍了一种使用冷冻画笔收获腺毛的方案。描述了一种使用液相色谱-质谱联用(2)分析进行广泛代谢物谱分析的工作流程,该流程适用于评估来自腺毛和其他植物组织的亲水性很强的类异戊二烯前体以及疏水性更强的代谢物。