College of Life and Environmental Sciences, Minzu University of China, Beijing, China.
Key Laboratory of Mass Spectrometry Imaging and Metabolomics (State Ethnic Affairs Commission), Centre for Imaging & Systems Biology, Minzu University of China, Beijing, China.
Plant Biotechnol J. 2023 Dec;21(12):2574-2584. doi: 10.1111/pbi.14154. Epub 2023 Aug 10.
A novel metabolomics analysis technique, termed matrix-assisted laser desorption/ionization mass spectrometry imaging-based plant tissue microarray (MALDI-MSI-PTMA), was successfully developed for high-throughput metabolite detection and imaging from plant tissues. This technique completely overcomes the disadvantage that metabolites cannot be accessible on an intact plant tissue due to the limitations of the special structures of plant cells (e.g. epicuticular wax, cuticle and cell wall) through homogenization of plant tissues, preparation of PTMA moulds and matrix spraying of PTMA sections. Our study shows several properties of MALDI-MSI-PTMA, including no need of sample separation and enrichment, high-throughput metabolite detection and imaging (>1000 samples per day), high-stability mass spectrometry data acquisition and imaging reconstruction and high reproducibility of data. This novel technique was successfully used to quickly evaluate the effects of two plant growth regulator treatments (i.e. 6-benzylaminopurine and N-phenyl-N'-1,2,3-thiadiazol-5-ylurea) on endogenous metabolite expression in plant tissue culture specimens of Dracocephalum rupestre Hance (D. rupestre). Intra-day and inter-day evaluations indicated that the metabolite data detected on PTMA sections had good reproducibility and stability. A total of 312 metabolite ion signals in leaves tissues of D. rupestre were detected, of which 228 metabolite ion signals were identified, they were composed of 122 primary metabolites, 90 secondary metabolites and 16 identified metabolites of unknown classification. The results demonstrated the advantages of MALDI-MSI-PTMA technique for enhancing the overall detection ability of metabolites in plant tissues, indicating that MALDI-MSI-PTMA has the potential to become a powerful routine practice for high-throughput metabolite study in plant science.
一种新型代谢组学分析技术，称为基于基质辅助激光解吸/电离质谱成像的植物组织微阵列（MALDI-MSI-PTMA），成功地开发用于高通量代谢物检测和成像从植物组织。该技术通过植物组织匀浆、PTMA 模具制备和 PTMA 切片基质喷涂，完全克服了由于植物细胞特殊结构（例如表皮蜡、角质层和细胞壁）的限制，使代谢物无法在完整的植物组织上获得的缺点。我们的研究展示了 MALDI-MSI-PTMA 的几个特性，包括无需样品分离和富集、高通量代谢物检测和成像（每天>1000 个样本）、高稳定性质谱数据采集和成像重建以及数据的高重现性。该新技术成功地用于快速评估两种植物生长调节剂处理（即 6-苄基氨基嘌呤和 N-苯基-N'-1,2,3-噻二唑-5-基脲）对Dracocephalum rupestre Hance（D. rupestre）植物组织培养标本中内源性代谢物表达的影响。日内和日间评估表明，PTMA 切片上检测到的代谢物数据具有良好的重现性和稳定性。在 D. rupestre 叶片组织中检测到 312 个代谢物离子信号，其中鉴定出 228 个代谢物离子信号，它们由 122 种初级代谢物、90 种次级代谢物和 16 种未知分类的鉴定代谢物组成。结果表明，MALDI-MSI-PTMA 技术用于增强植物组织中代谢物整体检测能力的优势，表明 MALDI-MSI-PTMA 有可能成为植物科学中高通量代谢物研究的有力常规实践。
