博落回根中苄基异喹啉生物碱合成、转运和积累的细胞类型特异性定位及分子机制研究

Benzylisoquinoline alkaloids (BIAs) are a type of plant secondary metabolites with high value for pharmaceutical utilization, but the urgent alkaloid-abundant plant resources limit its pharmaceutical development and manufacture. This project intends to investigate the cell-type specific metabolite profiling and genetic expressional profiling of BIAs from Macleaya Cordata root by the integrated modern technologies such as molecular mass spectrometry imaging (MSI), laser microdissection (LMD), liquid chromatography-mass spectrometry (LC-MS) combining with qRT-PCR on the basis of the previous research on the metabolomics, genomics and transcriptomics. The in situ imaging of BIAs can be present in the root cross sections at different cell types and growth stages using MSI. The different cell-type samples are cut by LMD, some of them are analyzed to acquire the target metabolite profilings of BIAs and their trends with time by LC-MS, temporal and spatial gene expressional profilings and gene regulation mechanism were elucidated using LMD-qRT-PCR technique. This study will contribute to the promotion of plant molecular breeding and metabolic engineering studies, and is expected to guide the development of alternative plant production systems.

苄基异喹啉生物碱（BIAs）是具有很高药用价值的植物次生代谢产物，但植物资源日趋紧张限制了其药用开发和利用。本项目以博落回根为研究材料，在前期代谢组、基因组和转录组研究基础上，将质谱成像（MSI）、激光显微切割（LMD）和qRT-PCR结合起来进行博落回根中BIAs的细胞类型特异性的代谢谱和基因表达谱研究。通过MSI对不同生长期博落回根中BIAs进行原位可视化的整体直观的代谢谱表征；利用LMD获取不同细胞类型样本，对样本中BIAs进行LC-MS/MS动态代谢谱分析；借助qRT-PCR对不同细胞类型样本中BIAs的合成、转运和积累相关基因的时空表达规律进行研究。将上述细胞类型特异性的BIAs代谢谱与基因表达谱的数据进行关联分析，揭示博落回根中细胞水平上BIAs的合成、转运和积累的细胞类型特异性定位及其分子机制。该研究将有助于促进植物分子育种或代谢工程研究，有望指导替代性生产体系的建立。