Bertic, Marko and Zimmer, Ina and Andres-Montaner, David and Rosenkranz, Maaria and Kangasjaervi, Jaakko and Schnitzler, Joerg-Peter and Ghirardo, Andrea (2023) Automatization of metabolite extraction for high-throughput metabolomics: case study on transgenic isoprene-emitting birch. TREE PHYSIOLOGY, 43 (10). pp. 1855-1869. ISSN 0829-318X, 1758-4469
Full text not available from this repository. (Request a copy)Abstract
Metabolomics studies are becoming increasingly common for understanding how plant metabolism responds to changes in environmental conditions, genetic manipulations and treatments. Despite the recent advances in metabolomics workflow, the sample preparation process still limits the high-throughput analysis in large-scale studies. Here, we present a highly flexible robotic system that integrates liquid handling, sonication, centrifugation, solvent evaporation and sample transfer processed in 96-well plates to automatize the metabolite extraction from leaf samples. We transferred an established manual extraction protocol performed to a robotic system, and with this, we show the optimization steps required to improve reproducibility and obtain comparable results in terms of extraction efficiency and accuracy. We then tested the robotic system to analyze the metabolomes of wild-type and four transgenic silver birch (Betula pendula Roth) lines under unstressed conditions. Birch trees were engineered to overexpress the poplar (Populus x canescens) isoprene synthase and to emit various amounts of isoprene. By fitting the different isoprene emission capacities of the transgenic trees with their leaf metabolomes, we observed an isoprene-dependent upregulation of some flavonoids and other secondary metabolites as well as carbohydrates, amino acid and lipid metabolites. By contrast, the disaccharide sucrose was found to be strongly negatively correlated to isoprene emission. The presented study illustrates the power of integrating robotics to increase the sample throughput, reduce human errors and labor time, and to ensure a fully controlled, monitored and standardized sample preparation procedure. Due to its modular and flexible structure, the robotic system can be easily adapted to other extraction protocols for the analysis of various tissues or plant species to achieve high-throughput metabolomics in plant research.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | SAMPLE PREPARATION; THYLAKOID MEMBRANES; MASS-SPECTROMETRY; TOBACCO PLANTS; EMISSION; PHOTOSYNTHESIS; PROTECTS; PLASMA; MODEL; MICROEXTRACTION; automation; sample preparation |
| Subjects: | 500 Science > 580 Botanical sciences |
| Divisions: | Biology, Preclinical Medicine > Institut für Pflanzenwissenschaften Biology, Preclinical Medicine > Institut für Pflanzenwissenschaften > Lehrstuhl für Ökologie und Naturschutzbiologie (Prof. Dr. Peter Poschlod) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 22 Apr 2024 13:20 |
| Last Modified: | 22 Apr 2024 13:20 |
| URI: | https://pred.uni-regensburg.de/id/eprint/62808 |
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