Qin Wei, Li Yongpeng, Peng Bowen, Liu Hang, Chen Tiantian, Yan Xin, Zhang Yaojie, Wang Chen, Yao Xinghao, Fu Xueqing, Li Ling, Tang Kexuan
Frontiers Science Center for Transformative Molecules, Joint International Research Laboratory of Metabolic and Developmental Sciences, Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China.
Front Plant Sci. 2022 Aug 22;13:985969. doi: 10.3389/fpls.2022.985969. eCollection 2022.
Trichomes, which are classified as glandular or non-glandular, are hair-like epidermal structures that are present on aerial parts of most plant species. Glandular secretory trichomes (GSTs) have the capacity to secrete and store specialized metabolites, which are widely used as natural pesticides, food additives, fragrance ingredients or pharmaceuticals. Isolating individual trichomes is an essential way for identifying trichome-specific gene functions and discovering novel metabolites. However, the isolation of trichomes is difficult and time-consuming. Here, we report a method to isolate the GSTs from leaf epidermis dispense with fixation using laser capture microdissection (LCM). In this study, 150 GSTs were captured efficiently from leaves and enriched for artemisinin measurement. UPLC analysis of microdissected samples indicated specific accumulation of secondary metabolites could be detected from a small number of GSTs. In addition, qRT-PCR revealed that the GST-specific structural genes involved in artemisinin biosynthesis pathway were highly expressed in GSTs. Taken together, we developed an efficient method to collect comparatively pure GSTs from unfixed leaved, so that the metabolites were relatively obtained intact. This method can be implemented in metabolomics research of purely specific plant cell populations and has the potential to discover novel secondary metabolites.
毛状体可分为腺毛和非腺毛，是大多数植物地上部分存在的毛发状表皮结构。腺毛分泌型毛状体（GSTs）具有分泌和储存特殊代谢物的能力，这些代谢物广泛用作天然杀虫剂、食品添加剂、香料成分或药物。分离单个毛状体是鉴定毛状体特异性基因功能和发现新代谢物的重要途径。然而，毛状体的分离既困难又耗时。在此，我们报告一种使用激光捕获显微切割（LCM）从叶片表皮分离GSTs而无需固定的方法。在本研究中，从叶片中高效捕获了150个GSTs，并富集用于青蒿素测定。对显微切割样品的超高效液相色谱（UPLC）分析表明，从少量GSTs中可检测到次生代谢物的特异性积累。此外，定量逆转录聚合酶链反应（qRT-PCR）显示，参与青蒿素生物合成途径的GST特异性结构基因在GSTs中高度表达。综上所述，我们开发了一种从未固定叶片中收集相对纯净GSTs的有效方法，从而使代谢物相对完整地获得。该方法可应用于纯特定植物细胞群体的代谢组学研究，并具有发现新的次生代谢物的潜力。
